No full-text available
To read the full-text of this research,
you can request a copy directly from the author.
Show and (sometimes) Tell: Identity Construction and the Affordances of Video-Conferencing
Abstract
This article provides a preliminary answer to exactly why video-conferencing is evaluated as better than traditional telephony for long-distance familial interaction by allocating analytical attention to the showing of objects during interaction. While it is acknowledged that ‘showing’ constitutes an interactive move less contingent on linguistic maturation, more importantly, the showing of objects, artefacts or entities during video-conferencing interactions exemplifies an agentive and volitional production of identity elements on behalf of young children. Thus, while some have pointed to shortcomings of conversation-like activities mediated by video-conferencing in favour for more activity-driven tasks, I make a case for drawing upon pre-existing components of the material surround as a means to more comprehensively and longitudinally engage younger children in video-conferencing interaction.
... This set-up required two types of recordings (screen-recording on the laptop, and an external camera that records an in-room view) and two researchers observing the interaction and taking notes. Geenen (2017) also investigated family interactions via videoconferencing using the same dataset reported in Norris (2016), and focused on the showing of objects, entities, and artefacts, particularly the ways in which this contributed to young children's agentive identity formation. Investigating showing as an interactive move, he elucidated how social actors explicitly acknowledged "the relationship between the objects, their interactive relevance and the frozen actions embedded in them" (p. ...
... Norris and Makboon (2015) had shown how identity markers were available as frozen actions in the print mode and objects backgrounded in social actors' attention/ awareness continuum in in-person interaction. Geenen (2017) extended the notion of frozen actions by demonstrating how identity markers were embedded and articulated in the actions of showing and telling as foregrounded in the social actors' attention/awareness. Norris and Pirini (2016) investigated everyday knowledge communication, specifically the higher-level actions of acknowledging knowledge, coordinating attention, and negotiating disagreement in dyadic teamwork via videoconferencing. ...
... Second, compared to Norris (2016) and Geenen (2017), the social actors in our work were alone in their physical spaces, which meant that there were no interactions with other actors beyond the screen. ...
Multimodal (inter)action analysis offers a powerful and robust methodology for the study of action and interaction between social actors, their environment, and the objects and tools within. Yet its implementation in the analysis of synchronous multimodal online data sets, e.g. (inter)actions via videoconferencing, is limited. Drawing on our research in understanding teacher-learner (inter)actions in instruction-giving fragments in synchronous multimodal online language lessons, we describe and illustrate the ways in which we adapted and extended some of the methodological and analytical tools. These include (1) the use of a grounded theory approach in delineating and identifying higher-level actions, (2) the embodiment and disembodiment of frozen actions, (3) electronic print mode, (4) semiotic lag, (5) semiotic (mis)alignment, (6) modal density (mis)alignment, and (7) how modal density can be achieved by brisk modal shifts in addition to through modal intensity and complexity. We conclude by a call for further educational research in online teaching platforms using the framework to have richer understandings of the (inter)actions between social actors with particular roles and identities (teachers-learners), their environment, and the objects and tools within, which bring their “own material properties, feel and techniques of use, affordances and limitations” (Chun, Dorothy, Richard Kern & Bryan Smith. 2016. Technology in language use, language teaching, and language learning. The Modern Language Journal 100. 64–80: 65).
... People tend to feel a degree of 'togetherness' that was superior to that of the traditional voice call. Visual access to both interlocutors and to physical space most likely play a role in these evaluations but Geenen (2017) argues that it is precisely the potential for the multimodal production of identity elements made possible by videoconferencing that may explain the comparatively positive evaluations. Based on a multimodal interaction analysis (Norris 2004, 2019; Geenen and Pirini 2020) of long-distance familial videoconferencing, Geenen argues that the parentally facilitated interactive practice of showing and sometimes talking about physical objects or entities in the physical surround allows young children to semi-autonomously produce identity elements in real-time which may not be possible through the medium of traditional telephony. ...
... While Geenen (2017) acknowledges this practice as 'collaborative', the precise nature of this multiparty collaboration is not detailed in any length. Specifically, contributions from other interlocutors who may not be directly visible are not detailed. ...
... interval. As Geenen (2017) argues, mediated actions can be very precisely inferred analytically by virtue of mediational residue which refers to the material results of mediated action. The lower-level action of object handling and more specifically using the mouse tracker to scroll while moving the cursor itself and be inferred from the visual alterations on screen. ...
FULL TEXT HERE: https://rfmv.fr/numeros/5/articles/11-visual-transcription/ The communication and language sciences have decidedly taken a multimodal turn and a proliferation of work in previously language-dominated fields is focusing on the contribution of non-verbal communicative modes in social interaction. While this has proven empirically and theoretically fruitful, it throws an additional kink in an already complicated issue: transcription practices and the interpretive and representational process accompanying said practices. In this article, we review and champion a methodological framework that provides analytical tools and a transcription protocol for the generation of visual transcripts. Visual transcription, we contest, is far more congruent with the multimodal ethos and we both detail the method while applying it to a data segment from a larger project investigation teamwork and collaborative problem-solving in dyadic videoconferencing interactions. The analytical focus on action coupled with the accompanying transcription method reveals complex and fluctuating distributions of attention and interactional awareness. Non-verbal actions reveal contributions to the task at hand which are not realized in talk and the analysis details an intricate ebb and flow of attentional orientations which are realized through non-verbal means.
... Mupinga, 2005) and the delivery of health care services (Wilcox, 2000). More recently, readily available infrastructure, devices, and software solutions have contributed to videoconferencing's becoming a social phenomenon (Geenen, 2017) by, for instance, helping members of long-distance families keep in touch (Follmer et al., 2010;King-O'Riain, 2015). Now video chatting increasingly replaces phone calls, especially amongst Millennials (Grech, 2019). ...
... After WCS became the main communication channel for many people at the same time, WCS emerged as a social technology that led to a new virtual togetherness. Affordances that long-distance families have long recognised (Geenen, 2017) were finally recognised by people who were suddenly restricted from meeting with each other. While personal interactions in this new context were expected, how this new virtual togetherness pertained to professional settings and the greater community in the form of virtual coffees, happy hours and the like seems unique. ...
... "zoombombing") and comes with new security and privacy risks. (Geenen, 2017). • Little evidence of pure WCS-mediated social meetings among workmates during work (e.g. ...
Regulations to contain the spread of COVID-19 have affected corporations, institutions, and individuals to a degree that most people have never seen before. Information systems researchers have initiated a discourse on information technology's role in helping people manage this situation. This study informs and substantiates this discourse based on an analysis of a rich dataset: Starting in March 2020, we collected about 3 million tweets that document people's use of web-conferencing systems (WCS) like Zoom during the COVID-19 crisis. Applying text-mining techniques to Twitter data and drawing on affordance theory, we derive five affordances of and five constraints to the use of WCS during the crisis. Based on our analysis, our argument is that WCS emerged as a social technology that led to a new virtual togetherness by facilitating access to everyday activities and contacts that were "locked away" because of COVID-19-mitigation efforts. We find that WCS facilitated encounters that could not have taken place otherwise and that WCS use led to a unique blending of various aspects of people's lives. Using our analysis, we derive implications and directions for future research to address existing constraints and realise the potentials of this period of forced digitalisation.
... One example of this approach comes from an ongoing study into video conferencing. The camera is ever-present in these interactions, and participants have learned to orientate to the camera to display their attention and participation, including visually introducing new topics by introducing objects (Geenen, 2017). The camera becomes a central material feature in the higher-level actions that social actors co-produce. ...
... They appear adept at tracking conversational topio and re-introducing said topics to newcomers who may have 'missed something'. The actions themselves, however, are more nuanced and do not always occur through the mode of spoken language as children draw upon a range of non-verbal modes like gesture and object handling to introduce topics (Geenen, 2017(Geenen, , 2018. Only a methodology which prioritises the pragmatics of interaction (Clark, 2014) can help accurately deduce when and to what extent communicative aptitudes emerge. ...
Multimodal (Inter)action Analysis was developed to study social interaction based upon the theoretical notion of mediated action. Building on this core concept, Multimodal (Inter)action Analysis includes several theoretical/methodological tools. These tools facilitate analysis which moves flexibly between micro-level moments of interaction and macro-level practices and discourses. In this chapter, the application of mediated action to multimodal analysis is discussed, before the central theoretical/methodological tools are introduced. Tight links are made between the tools used in Multimodal (Inter)action Analysis and the core theoretical tenets, to support robust multimodal interaction research.
... Since a mediated action always involves a social actor, acting with and through mediational means, a central form of data is video recordings of social actors. Some examples of settings where researchers have used this framework include classroom interactions (Norris, 2014), family video-conferencing interactions (Geenen, 2017) (see Sample Study 28.1), management of attention/awareness in high school tutoring sessions (Pirini, 2014b), sign language in classrooms (Tapio, 2014), construction of ethnic identity within adult learning environments (Matelau, 2014), and the structure of lectures (Bernad-Mechó, 2017). ...
... Sample Study 28.1 walks readers through an example of a Multimodal (Inter)action Analysis studying family-child interaction over Skype. Sample Study 28.1 Geenen, J. (2017). Show and (sometimes) tell: Identity construction and the affordances of video-conferencing. ...
This chapter introduces the five most prominent approaches to multimodal data analysis: Multimodal (Inter)action Analysis, Mediated Discourse Analysis, Systemic Functional Multimodal Discourse Analysis, Social Semiotics, and Multimodal Conversation Analysis. The chapter begins by discussing the origin of each approach, methods of data collection and analysis, and embedded theoretical foundations. The theoretical differences between the approaches are then examined with a focus on the way each approach treats the individual, the artefact, and the notion of mode during analysis. With the variety of available approaches to multimodal data analysis, it is important that researchers link data collection and analysis to coherent theoretical underpinnings.
... Multimodal (Inter)action analysis has already proved quite valuable in unravelling some of the complexities which emerge through new technologically-mediated forms of social interaction. In the fractured interactional ecologies which emerge when communication is complexly mediated by new technologies, non-verbal modes take on a new importance and appear to be pivotal for exemplifying divergence in stance or disagreement (Norris and Pirini 2017), can be vital for the production of identity (Geenen 2017) and can be exploited to help facilitate smoother interactions with pre-verbal children and toddlers (Geenen 2018(Geenen , 2020. Other work has shown how non-verbal actions can be pivotal to accomplishing collaborative tasks via video-conferencing technology (Geenen et al. 2021) but that the distribution of attention and the interactional demands can also result in miscommunication and misunderstanding (Norris and Geenen 2022). ...
... Multimodal (inter)action analysis is thus a coherent and comprehensive research framework for the analysis of qualitative video-based data. 2 All the pieces in this framework fit together (Norris 2012;Pirini 2014b), allowing the researcher to build a coherent picture of whatever human action, interaction or identity is being studied. In this way, we have made strides in examining space and place or children's acquisition (Geenen 2013;Geenen 2017Geenen , 2018; identity (Norris 2005(Norris , 2007(Norris , 2008(Norris , 2011Norris and Makboon 2015;Matelau-Doherty and Norris 2021); video conferences (Norris 2017a;Norris and Pirini 2017); business coaching, high school tutoring and intersubjectivity (Pirini 2013(Pirini , 2014a(Pirini , 2016, to name but a few areas in which the framework has been used. What we at the AUT Multimodal Research Centre are finding is that with a coherent framework such as MIA, there is much potential to discover new insight and knowledge about any kind of human action, interaction, and identity. ...
This paper presents a concise introduction to Multimodal (inter)action analysis (MIA), which began to be developed in the early 2000s in tandem with technological advances for visual qualitative research. By now, MIA has grown into a fully-fledged research framework, including multimodal philosophy, theory, method and methodology for the study of human action, interaction and identity. With systematic phases from data collection to transcription (including transcription conventions) and data analysis, this framework allows researchers to work in a data-driven and replicable manner moving past common interpretive paradigms (Norris 2019, 2020).
Red Chinese movies depict heroic individuals who make significant contributions to the nation, and these films play a vital role in constructing the national image of new China. This paper explores the images of heroic individuals in red Chinese movies using multimodal action theory. The findings indicate that these individuals are characterized by their selfless sacrifices for their country, fearless perseverance, embodiment of international communist ideals, and unwavering devotion and warmth. The study also reveals that facial expressions, body movements, and language serve as critical mediational means in shaping these characters, while other forms of mediation are underutilized. Recommendations include employing more mediational means to create more nuanced and complex characters in future red Chinese movies.
Purpose
The purpose of this study is to examine the role of videoconferencing technologies for mediating and transforming emotional experiences in virtual context.
Design/methodology/approach
Drawing on empirical data of video conferencing experiences, this study identifies different constitutive relations with technology through which actors cope with actual or potential anxieties in virtual meetings. It draws on the phenomenological-existential tradition (Sartre and Merleau-Ponty) and on an interpretive phenomenological analysis (IPA) to conceptualize and illustrate the role of affective affordances in virtual settings.
Findings
The study identifies four different body–technology–other relations that provide different action possibilities, both disclosing and concealing, for navigating emotional experiences in virtual encounters of mutual gazing. These findings offer insights into the anatomy of virtual emotions and provide explanations on the nature of Zoom fatigue (interactive exhaustion) and heightened feelings of self-consciousness resulting from video conferencing interactions.
Originality/value
This paper builds on and extends current scholarship on technological affordances, as well as emotions, to suggest that technologies also afford different tactics for navigating emotional experiences. Thus, this paper proposes the notion of affective affordance that can expand current information system (IS) and organization studies (OS) scholarship in important ways. The focus is on videoconference technologies and meetings that have received little research attention and even less so from a perspective on emotions. Importantly, the paper offers nuanced insights that can advance current research discourse on the relationships between technology, human body and emotions.
This thesis examines the construction of hybrid and fluid ethnic identity elements as produced by Māori and Pacific female artists. Ethnic identity studies within New Zealand reveal different types of ethnic identities, and although there is research being conducted into hybrid and fluid Māori and Pacific identity elements, no studies have been done examining the construction of these identities through mediated action. This thesis attempts to fill this gap.
Using video ethnography and socio-linguistic interviews, data were collected and analysed utilising multimodal (inter)action analysis (MIA) as the theoretical and methodological framework. Vertical identity production and site of engagement are analytical tools within MIA that allow for the study of the intersection between discourses and mediated actions performed by social actors. These analytical tools were applied to interview and video transcripts selected from the data, following a systematic process of data cataloguing.
Analysis of the data is presented in three chapters which show the ethnic and creative identity production of the participants as constructed through the central, intermediary and outer layers of discourse. The first analysis chapter demonstrates the way the participants create art by blending traditional and contemporary features and diverse knowledge, in turn constructing their immediate ethnic and creative identity elements. This analysis is compared to the way the participants verbalise these identity elements within their interviews. The second analysis chapter examines the way experiences of exclusion and inclusion from within their networks shape their continuous ethnic and
creative identity elements. The third analysis chapter explores moments of exclusion and inclusion but within larger communities such as mainstream New Zealand, and their ethnic communities. It also illustrates the way in which the participants’ art creates inclusion and shapes the general ethnic and creative identity development of other social actors.
Following this, wider discourses and practices are examined using the site of engagement as the analytical tool. This chapter demonstrates the way in which wider discourses such as colonial, superiority/inferiority and racism discourse intersect with practices such as superiority/inferiority, gratitude, and marginalisation and with the mediated actions performed by the participants. This analysis highlights the negative impact these discourses and practices can have on ethnic identity construction for Māori and Pacific social actors. To this end, numerous recommendations are made within the conclusion with the intention of changing these wider discourses and practices.
This thesis contributes to knowledge in the area of Māori and Pacific identity studies by utilising multimodal (inter)action analysis to study identity production. It also contributes to the theoretical and methodological framework of multimodal (inter)action analysis.
This paper examines the use of video chat (VC) with a focus on expectations and construction of attention. It is based on micro analyses of recorded VC sessions (gathered between 2013 and 2015) and thematic analysis of 29 semi-structured interviews about VC practices (conducted in 2014 and 2015). Building on multimodal (inter)action analysis (Norris, S. (2004). Analysing multimodal interaction: a methdological framework. Routledge, Norris, S. (2016). Concepts in multimodal discourse analysis with examples from video conferencing. Yearbook of the Poznan Linguistic Meeting 2: 141–165) and key concepts from nexus analysis (Scollon, R. and Scollon, S.W. (2004). Nexus Analysis: Discourse and the emerging internet. Routledge), I examine how focused attention is constructed in VCs and how these practices are shaped by experiences with other forms of communication. I demonstrate that unlike other forms of distance communication, typical VC encounters require a full investment of attention. This can be formulated as an interactional maxim: focus your attention on the VC interaction . I discuss how other activities can be interwoven with a VC and examine the exceptional practice of lapsed VC encounters (previously open connections or always-on video). I argue that participants display an orientation towards the maxim when pursuing other courses of action, and that lapsed encounters operate under a different value system than typical focused VC encounters. Finally, I reason that VC is reserved for close relationships because of the required investment of attention.
Multimodal interaction analysis is a holistic methodological framework that allows the analyst to integrate the verbal with the nonverbal, and to integrate these with material objects and the environment as they are being used by individuals acting and interacting in the world. In short, multimodal interaction analysis allows a researcher to study real people interacting with others, with technology, and with the environment. The point of view taken in multimodal interaction analysis is that all actions in fact are interactions and that all of these (inter)actions are linked to people (referred to as “social actors” in multimodal interaction analysis)—no matter whether you are investigating the real or the virtual world, someone buying ice cream, or using a software program. Multimodal interaction analysis is carried out through a series of analytical phases and steps. Researchers utilize multimodal transcription conventions and analytical tools, which are theoretically founded, allowing for replicability of analyses and reliability of findings. Multimodal interaction analysis allows the integration of all communicative modes, where communicative modes are defined as systems of mediated actions. In this view, all modes of communication together build one system of communication, which comes about through actions and interactions that people produce. This coherent framework and process of analysis can be taught and learned and is proving useful for research examining what people do, how they communicate, and how they interact.
Hypercortisolism is one of the most commonly diagnosed endocrinopathies in dogs, and new targeted medical treatment options are desirable. Steroidogenic factor-1 (SF-1), an orphan nuclear hormone receptor, is a key regulator of adrenal steroidogenesis, development, and growth. In pituitary-dependent hypercortisolism (PDH), high plasma ACTH concentrations increase the transcriptional activity of SF-1. In adrenal-dependent hypercortisolism, SF-1 expression is significantly greater in dogs with recurrence after adrenalectomy than in those without recurrence. Inhibition of SF-1 could therefore be an interesting treatment option in canine spontaneous hypercortisolism. We determined the effects of 3 SF-1 inverse agonists, compounds IsoQ A, #31, and #32, on cortisol production, on the messenger RNA (mRNA) expression of steroidogenic enzymes and SFs, and on cell viability, in primary adrenocortical cell cultures of 8 normal adrenal glands and of 3 cortisol-secreting adrenocortical tumors (ATs). To mimic PDH, the normal adrenocortical cell cultures were stimulated with ACTH. The results show that only compound #31 inhibited cortisol production and SF-1 target gene expression in non-ACTH-stimulated and ACTH-stimulated normal adrenocortical cells but did not affect cell viability. In the AT cell cultures, the effects of #31 on cortisol production and target gene expression were variable, possibly caused by a difference in the SF-1 mRNA expressions of the primary tumors. In conclusion, inhibition of SF-1 activity shows much promise as a future treatment for canine hypercortisolism.
Background
Circulating cell-free miRNAs have emerged as promising minimally-invasive biomarkers for early detection, prognosis and monitoring of cancer. They can exist in the bloodstream incorporated into extracellular vesicles (EVs) and ribonucleoprotein complexes. However, it is still debated if EVs contain biologically meaningful amounts of miRNAs and may provide a better source of miRNA biomarkers than whole plasma. The aim of this study was to systematically compare the diagnostic potential of prostate cancer-associated miRNAs in whole plasma and in plasma EVs.
Methods
RNA was isolated from whole plasma and plasma EV samples from a well characterised cohort of 50 patient with prostate cancer (PC) and 22 patients with benign prostatic hyperplasia (BPH). Nine miRNAs known to have a diagnostic potential for PC in cell-free blood were quantified by RT-qPCR and the relative quantities were compared between patients with PC and BPH and between PC patients with Gleason score ≥ 8 and ≤6.
Results
Only a small fraction of the total cell-free miRNA was recovered from the plasma EVs, however the EV-incorporated and whole plasma cell-free miRNA profiles were clearly different. Four of the miRNAs analysed showed a diagnostic potential in our patient cohort. MiR-375 could differentiate between PC and BPH patients when analysed in the whole plasma, while miR-200c-3p and miR-21-5p performed better when analysed in plasma EVs. EV-incorporated but not whole plasma Let-7a-5p level could distinguish PC patients with Gleason score ≥ 8 vs ≤6.
Conclusions
This study demonstrates that for some miRNA biomarkers EVs provide a more consistent source of RNA than whole plasma, while other miRNAs show better diagnostic performance when tested in the whole plasma.
Sphingosine kinase 1 (Sphk1) is a highly conserved lipid kinase that phosphorylates sphingosine to form sphingosine-1-phosphate (S1P). Growing studies have demonstrated that Sphk1 is overexpressed in various types of solid cancers and can be induced by growth factors, cytokines, and carcinogens, leading to the increase of S1P production. Subsequently, the increased Sphk1/S1P facilitates cancer cell proliferation, mobility, angiogenesis, invasion, and metastasis. Therefore, Sphk1/S1P signaling plays oncogenic roles. This review summarizes the features of Sphk1/S1P signaling and their functions in colorectal cancer cell growth, tumorigenesis, and metastasis, as well as the possible underlying mechanisms.
Background:
Detection of circulating cell-free mRNA serves as noninvasive tools for cancer diagnosis. As an oncofetal protein, HMGA2 (high mobility group AT-hook 2) is upregulated in colorectal cancer (CRC) tissues. However, it is not clear whether the increased levels of circulating cell-free HMGA2 mRNA functions as potential biomarkers for improved diagnosis of CRC.
Methods:
To assess its clinical significance in diagnosis and prediction, we evaluated serum levels of circulating HMGA2 mRNA in CRC patients and in healthy controls. In this study, 83 CRC patients and 11 normal controls were enrolled in this study. We used real-time quantitative reverse transcription-PCR to evaluate the plasma mRNA levels of HMGA2 and analyze the correlation between their expression and clinicopathologic characteristics.
Results:
We found that the levels of HMGA2 mRNA were significantly higher in CRC patients compared with healthy volunteers. The patients with right-sided CRC, colon cancer, positive nerve infiltration, positive vascular invasion, negative microsatellite instability (MSI), and increasing in serum carbohydrate antigen (CA) 199 had higher levels of plasma HMGA2 mRNA. A strong positive correlation between circulating cell-free HMGA2 mRNA and CA199 level in serum was found in our study. Furthermore, statistical analysis revealed that levels of HMGA2 mRNA in plasma and in tumors were strictly correlated.
Conclusions:
Collectively, our data suggested that cell-free HMGA2 mRNA in plasma might function as a novel diagnostic marker for CRC.
Since their discovery and the advent of RNA interference, microRNAs have drawn enormous attention because of their ubiquitous involvement in cellular pathways from life to death, from metabolism to communication. It is also widely accepted that they possess an undeniable role in cancer both as tumor suppressors and tumor promoters modulating cell proliferation and migration, epithelial-mesenchymal transition and tumor cell invasion and metastasis. Moreover, microRNAs can even affect the tumor surrounding environment influencing angiogenesis and immune system activation and recruitment. The tight association of microRNAs with several cancer-related processes makes them undoubtedly connected to the effect of specific cancer drugs inducing either resistance or sensitization. In this context, personalized medicine through microRNAs arose recently with the discovery of single nucleotide polymorphisms in the target binding sites, in the sequence of the microRNA itself or in microRNA biogenesis related genes, increasing risk, susceptibility and progression of multiple types of cancer in different sets of the population. The depicted scenario implies that the overall variation displayed by these small non-coding RNAs have an impact on patient-specific pharmacokinetics and pharmacodynamics of cancer drugs, pushing on a rising need of personalized treatment. Indeed, microRNAs from either tissues or liquid biopsies are also extensively studied as valuable biomarkers for disease early recognition, progression and prognosis. Despite microRNAs being intensively studied in recent years, a comprehensive review describing these topics all in one is missing. Here we report an up-to-date and critical summary of microRNAs as tools for better understanding personalized cancer biogenesis, evolution, diagnosis and treatment.
miRNA is a short non-coding RNA that can influence mRNA processing at the post-transcriptional level. A large number of miRNAs have been found in virtually all species so far, and these small molecules play an important role in many different physiological processes and various pathologic conditions, such as cell metabolism, cancer, autoimmune disease, and diabetes mellitus. T2D arises from a dysregulated response to the elevated glucose level in the circulation. The prevalence of T2D has increased dramatically in all age groups, and T2D in older adults is associated with more T2D complications and higher mortality. Despite the existing findings describing the pathological mechanism, T2D pathology is more complex and the pathophysiology of the disease is still not fully elucidated. In this review, we summarize the current understanding of miRNA-mediated modulation of gene expression in T2D pathogenesis, as well as related signaling pathways, and insight into the important role of miRNA in various T2D complications. Furthermore, the potential therapeutic value of miRNA for T2D patients is also discussed in detail.
A lo largo de los últimos 20 años, el gran avance del desarrollo tecnológico ha traído consigo un importante y profundo conocimiento de las bases genéticas y moleculares del ser humano. Las bases sobre las que asentaban los conocimientos en torno a la genética clínica y las relaciones entre causa genética y el desarrollo de enfermedades hereditarias se han visto modificados de forma trascendental. En este ámbito, los descubrimientos en el campo de la genética clínica se han visto orientados hacia conceptos de heterogeneidad genética y fenotípica, de forma que una misma patología hereditaria actualmente se sabe que podría estar ocasionada por alteraciones en diferentes genes y simultáneamente, variantes en el mismo gen podrían desencadenar fenotipos de gran diversidad.
En este nuevo panorama, las metodologías de análisis masivo de genes así como las implementaciones bioinformáticas necesarias para la interpretación de las alteraciones identificadas, resultan de elevada importancia para poder alcanzar el diagnóstico certero de una enfermedad hereditaria.
Esta misma profundización en el conocimiento genético, ha identificado nuevos ámbitos de estudio que anteriormente se desconocían. La farmacogenética ha emergido como una disciplina de vital relevancia debido a que los perfiles genéticos propios del individuo influyen en la respuesta farmacológica final en términos de seguridad y efectividad. Además ha dejado al descubierto requerimientos adicionales para los estudios genéticos, como parámetros de sensibilidad, límites de detección y tiempos de respuesta, particularmente relevantes en el ámbito oncológico.
Adicionalmente las matrices biológicas sustrato de los estudios genéticos han evolucionado junto con el avance científico. La identificación de microARNs y la existencia de ADN tumoral circulante (ctADN) como biomarcadores plasmáticos, no invasivos que han mostrado en recientes estudios su potencial funcionalidad como indicadores en el screening y monitorización de progresión de enfermedad.
En términos de costo-efectividad, nuestros esfuerzos deberían dirigirse los hacia el diagnóstico precoz de enfermedades de alta heterogeneidad genética y en la rápida detección de recidivas en enfermedad neoplásica. Ambas situaciones incrementan las posibilidades de éxito del tratamiento farmacológico en el paciente y su familia, y por tanto contribuyen a aumentar la supervivencia del paciente y mejorar su calidad de vida.
On the grounds that miRNAs present in the blood of prostate cancer (PCa) patients are released in the growth medium by PCa cells, it is conceivable that PCa cells resistant to docetaxel (DCT) (DCTR) will release miRNAs that may be found in PCa patients under DCT therapy if resistant PCa cells appear. We isolated DCTR clones respectively from 22Rv1 and DU-145 PCa cell lines and performed through next-generation sequencing (NGS) the miRNAs profiles of the released miRNAs. The analysis of the NGS data identified 105 and 1 miRNAs which were differentially released in the growth medium of the 22Rv1/DCTR and DU-145/DCTR clones, respectively. Using additional filters, we selected 12 and 1 miRNA more released by all 22Rv1/DCTR and DU-145/DCTR clones, respectively. Moreover, we showed that 6 of them were more represented in the growth medium of the DCTR cells than the ones of DCT-treated cells. We speculated that they have the pre-requisite to be tested as predictive biomarkers of the DCT resistance in PCa patients under DCT therapy. We propose the utilization of clones resistant to a given drug as in vitro model to identify the differentially released miRNAs, which in perspective could be tested as predictive biomarkers of drug resistance in tumor patients under therapy.
Exosomes are vesicles of endocytic origin released by many cells. These vesicles can mediate communication between cells, facilitating processes such as antigen presentation. Here, we show that exosomes from a mouse and a human mast cell line (MC/9 and HMC-1, respectively), as well as primary bone marrow-derived mouse mast cells, contain RNA. Microarray assessments revealed the presence of mRNA from approximately 1300 genes, many of which are not present in the cytoplasm of the donor cell. In vitro translation proved that the exosome mRNAs were functional. Quality control RNA analysis of total RNA derived from exosomes also revealed presence of small RNAs, including microRNAs. The RNA from mast cell exosomes is transferable to other mouse and human mast cells. After transfer of mouse exosomal RNA to human mast cells, new mouse proteins were found in the recipient cells, indicating that transferred exosomal mRNA can be translated after entering another cell. In summary, we show that exosomes contain both mRNA and microRNA, which can be delivered to another cell, and can be functional in this new location. We propose that this RNA is called " exosomal shuttle RNA " (esRNA). Exosomes are small (50–90 nm) membrane vesicles of endocytic origin that are released into the extracellular environment on fusion of multivesicular bodies (MVB) with the plasma membrane 1. Many cells have the capacity to release exosomes, including reticulo-cytes 2 , dendritic cells 3 , B cells 4 , T cells 5 , mast cells 6 , epithelial cells 7 and tumour cells 8. The functions of exosomes are not completely understood, although it has been shown that exosomes can participate in the signalling events contributing to antigen presentation to T cells 4 and the development of tolerance 9. Several mechanisms have been hypothesized describing the interactions of exosomes and recipient cells. Exosomes can bind to cells through recep-tor–ligand interactions, similar to cell–cell communication mediating , for example, antigen presentation 4. Alternatively, exosomes could putatively attach or fuse with the target-cell membrane, delivering exosomal surface proteins and perhaps cytoplasm to the recipient cell 10,11. Finally, exosomes may also be internalized by the recipient cells by mechanisms such as endocytosis 12. Exosomes were isolated from a mast-cell line (MC/9), primary bone marrow-derived mast cells (BMMC) and a human mast-cell line (HMC-1) through a series of microfiltration and ultracentrifugation steps modified from what has been previously described 4. To confirm that the structures studied indeed are exosomes, they were examined by electron microscopy (Fig. 1a), flow cytometric analysis (FACS; Fig. 1b), and proteomic analysis (see Supplementary Information, Table S1). The electron micrographs of the exosomes revealed rounded structures with a size of approximately 50–80 nm, similar to previously described exo-somes 4,13–15. The identity of the studied vesicles was further confirmed as exosomes by FACS analysis (Fig. 1b), which show the presence of the surface protein CD63 — a commonly used marker of exosomes. Finally, extensive protein analysis of the MC/9 exosomes was performed on multiple samples using LC-MS/MS technology. A total of 271 proteins were identified (see Supplementary Information, Table S1) from three preparations of the isolated vesicles, of which 47 proteins were present in all three samples. More importantly, a large number of the proteins found in the preparations were the same as proteins previously identified in exosomes produced by other cells (that is, exosomes derived from intestinal epithelial cells, urine, dendritic cells, microglia, melanoma, T-cells and B-cells). In particular, 60% of the 47 proteins found in all samples of mast-cell exosomes have been previously found in other types of exosomes. Moreover, 39% of the 271 total proteins found in the analysed exosome samples have also been previously found in other types of exosomes. Thus, the electron microscopy, the FACS, and the detailed protein analyses each provided significant evidence in favour of the identification of the isolated vesicles as exosomes. The presence of nucleic acids was examined in exosomes derived from MC/9, BMMC and HMC-1 cells to define a potential mechanism by which exosomes may mediate cell–cell communication. These assessments showed that isolated exosomes contain no DNA (see Supplementary Information, Fig. S1). However, substantial amounts of RNA were detected by agarose gel electrophoresis, spectrophotometry
Exosomes are one type of membrane vesicles secreted into extracellular space by most types of cells. In addition to performing many biological functions particularly in cell-cell communication, cumulative evidence has suggested that several biological entities in exosomes like proteins and microRNAs are closely associated with the pathogenesis of most human malignancies and they may serve as invaluable biomarkers for disease diagnosis, prognosis, and therapy. This provides a commanding impetus and growing demands for simple, efficient, and affordable techniques to isolate exosomes. Capitalizing on the physicochemical and biochemical properties of exosomes, a number of techniques have been developed for the isolation of exosomes. This article summarizes the advances in exosome isolation techniques with an emphasis on their isolation mechanism, performance, challenges, and prospects. We hope that this article will provide an overview of exosome isolation techniques, opening up new perspectives towards the development more innovative strategies and devices for more time saving, cost effective, and efficient isolations of exosomes from a wide range of biological matrices.
MicroRNAs (miRNAs) hold promise as biomarkers for a variety of disease processes and for determining cell differentiation. These short RNA species are robust, survive harsh treatment and storage conditions and may be extracted from blood and tissue. Pre-analytical variables are critical confounders in the analysis of miRNAs: we elucidate these and identify best practices for minimizing sample variation in blood and tissue specimens. Pre-analytical variables addressed include patient-intrinsic variation, time and temperature from sample collection to storage or processing, processing methods, contamination by cells and blood components, RNA extraction method, normalization, and storage time/conditions. For circulating miRNAs, hemolysis and blood cell contamination significantly affect profiles; samples should be processed within 2 h of collection; ethylene diamine tetraacetic acid (EDTA) is preferred while heparin should be avoided; samples should be “double spun” or filtered; room temperature or 4 °C storage for up to 24 h is preferred; miRNAs are stable for at least 1 year at –20 °C or –80 °C. For tissue-based analysis, warm ischemic time should be <1 h; cold ischemic time (4 °C) <24 h; common fixative used for all specimens; formalin fix up to 72 h prior to processing; enrich for cells of interest; validate candidate biomarkers with in situ visualization. Most importantly, all specimen types should have standard and common workflows with careful documentation of relevant pre-analytical variables.
Background and aim
The potential of microRNAs (miRNA) as non-invasive diagnostic, prognostic, and predictive biomarkers, as well as therapeutic targets, has recently been recognized. Previous studies have highlighted the importance of consistency in the methodology used, but to our knowledge, no study has described the methodology of sample preparation and storage systematically with respect to miRNAs as blood biomarkers. The aim of this study was to investigate the stability of miRNAs in blood under various relevant clinical and research conditions: different collection tubes, storage at different temperatures, physical disturbance, as well as serial freeze-thaw cycles.
Methods
Blood samples were collected from 12 healthy donors into different collection tubes containing anticoagulants, including EDTA, citrate and lithium-heparin, as well as into serum collection tubes. MiRNA stability was evaluated by measuring expression changes of miR-1, miR-21 and miR-29b at different conditions: varying processing time of whole blood (up to 72 hours (h)), long-term storage (9 months at -80°C), physical disturbance (1 and 8 h), as well as in a series of freeze/thaw cycles (1 and 4 times).
Results
Different collection tubes revealed comparable concentrations of miR-1, miR-21 and miR-29b. Tubes with lithium-heparin were found unsuitable for miRNA quantification. MiRNA levels were stable for at least 24 h at room temperature in whole blood, while separated fractions did show alterations within 24 h. There were significant changes in the miR-21 and miR-29b levels after 72 h incubation of whole blood at room temperature (p<0.01 for both). Both miR-1 and miR-21 showed decreased levels after physical disturbance for 8 h in separated plasma and miR-1 in serum whole blood, while after 1 h of disturbance no changes were observed. Storage of samples at -80°C extended the miRNA stability remarkably, however, miRNA levels in long-term stored (9 months) whole blood samples were significantly changed, which is in contrast to the plasma samples, where miR-21 or miR-29b levels were found to be stable. Repetitive (n = 4) freeze-thaw cycles resulted in a significant reduction of miRNA concentration both in plasma and serum samples.
Conclusion
This study highlights the importance of proper and systematic sample collection and preparation when measuring circulating miRNAs, e.g., in context of clinical trials. We demonstrated that the type of collection tubes, preparation, handling and storage of samples should be standardized to avoid confounding variables influencing the results.
Exosomal miRNAs are proposed as excellent candidate biomarkers for clinical applications. However, little is known about their potential roles as prognostic biomarkers in lung cancer. In this study, we explored the prognostic value of plasma exosomal microRNAs (miRNAs) for non-small-cell lung cancer (NSCLC). Using a quantitative polymerase chain reaction (qPCR) array panel, we analyzed 84 plasma exosomal miRNAs in 10 lung adenocarcinoma patients and 10 matched healthy controls. The qPCR array showed 30 aberrantly expressed exosomal miRNAs. Nine candidate miRNAs were selected based on differential expression and previous reports for further evaluating their prognostic roles in 196 NSCLC patients. Elevated levels of exosomal miR-23b-3p, miR-10b-5p and miR-21-5p were independently associated with poor overall survival (with hazard ratio [95% confidence interval]: 2.42 (1.45 - 4.04), P = 0.001; 2.22 (1.18 - 4.16), P = 0.013; 2.12 (1.28 - 3.49), P = 0.003, respectively). When compared to the clinical prognostic variables only model, adding the three exosomal miRNA signatures significantly improved survival predictive accuracy with an increase of time-dependent area under the receiver operating characteristic curve from 0.88 to 0.91 (P=0.015). Our results indicated that plasma exosomal miR-23b-3p, miR-10b-5p and miR-21-5p are promising non-invasive prognostic biomarkers of NSCLC.
Circulating RNAs, especially microRNAs (miRNAs), have recently emerged as non-invasive disease biomarkers. Despite enthusiasm and numerous reports on disease-associated circulating miRNAs, currently there is no circulating miRNA-based diagnostic in use. In addition, there are many contradictory reports on the concentration changes of specific miRNA in circulation. Here we review the impact of various technical and non-technical factors related to circulating miRNA measurement and elucidate the importance of having a general guideline for sample preparation and concentration measurement in studying circulating RNA.
Introduction
Diagnosis of prostate cancer (PCa) is based on prostate biopsy that is performed when prostate specific antigen (PSA) is persistently altered over time and/or abnormal digital rectal examination is found. Serum PSA levels increase in both PCa and benign prostatic hyperplasia, leading to an increased number of unnecessary biopsies. There is an urgent need to unravel PCa-specific molecular signatures.
Patients and methods
This study aimed at characterizing a panel of circulating micro RNAs (miRNAs) that could distinguish PCa from benign prostatic hyperplasia in a population of age-matched patients with increased PSA levels. Both miRNAs targeting genes involved in PCa onset and miRNAs whose role in PCa has been highlighted in other studies were included. For this purpose, let-7c, let-7e, let-7i, miR-26a-5p, miR-26b-5p, miR-24-3p, miR-23b-3p, miR-27-b-3p, miR-106a-5p, miR-20b-5p, miR-18b-5p, miR-19b-2-5p, miR-363-3p, miR-497, miR-195, miR-25-3p, miR-30c-5p, miR-622, miR-874-3p, miR-346 and miR-940 were assayed through real-time PCR in 64 patients with PCa and compared with 60 patients with benign prostatic hyperplasia. The ability of miRNAs to predict the stage of disease was also analyzed.
Results
Let-7c, let-7e, let-7i, miR-26a-5p, miR-26b-5p, miR-18b-5p and miR-25-3p were able to discriminate patients with PCa from those harboring benign prostatic hyperplasia, both presenting altered PSA levels (>3 ng/mL). MiR-25-3p and miR-18b-5p showed the highest sensitivity and specificity to predict PCa, respectively. The combination of these two miRNAs improved the overall sensitivity. A correlation between pathological Gleason score and miRNA expression levels was reported; miR-363-3p, miR-26a-5p, miR-26b-5p, miR-106a-5p, miR-18b-5p, miR-25-3p and let-7i decreased in expression concomitantly with an increase in malignancy.
Conclusion
This study confirms serum miRNAs to be reliable candidates for the development of minimally invasive biomarkers for the diagnosis and prognosis of PCa, particularly in those cases where PSA acts as a flawed marker.
Evidence has shown that microRNAs are widely implicated as indispensable components of tumor suppressive and oncogenic pathways in human cancers. Thus, identification of microRNA targets and their relevant pathways will contribute to the development of microRNA-based therapeutics. The forkhead box transcription factors regulate numerous processes including cell cycle progression, metabolism, metastasis and angiogenesis, thereby facilitating tumor initiation and progression. A complex network of protein and non-coding RNAs mediates the expression and activity of forkhead box transcription factors. In this review, we summarize the current knowledge and concepts concerning the involvement of microRNAs and forkhead box transcription factors and describe the roles of microRNAs-forkhead box axis in various disease states including tumor initiation and progression. Additionally, we describe some of the technical challenges in the use of the microRNA-forkhead box signaling pathway in cancer treatment.
Various vaccine treatments against metastatic colorectal cancer have been developed and applied. However, to improve the efficacy of immunotherapy, biomarkers that can predict the effects are needed. It has been reported that various microRNAs (miRNAs) in peripheral blood may be useful as non-invasive biomarkers. In this study, miRNAs influencing the efficacy of vaccine treatment were screened for in a microarray analysis of 13 plasma samples that were obtained from patients prior to vaccine treatment. To validate the screening results, real-time RT-PCR was performed using 93 plasma samples obtained from patients prior to vaccine treatment. Four candidate miRNAs were selected according to the results of the comprehensive analysis of miRNA expression, which were ranked using the Fisher criterion and the absolute value of the log2 ratio in the screening analysis. The validation analysis showed that in the HLA-A*2402‑matched patient group (vaccine-treated group), patients with a high expression of plasma miR-6826 had a poorer prognosis than those with a low expression (P=0.048). In contrast, in the HLA-A*2402-unmatched patient group (control group), there was no difference between the patients with high or low plasma miR-6826 expression (P=0.168). Similar results were obtained in the analysis of miR-6875 (P=0.029 and P=0.754, respectively). Moreover, multivariate analysis of the Cox regression model indicated that the expression of miR-6826 was the most significant predictor for overall survival (P=0.003, hazard ratio, 3.670). In conclusion, plasma miR-6826 and miR-6875 may be predictive biomarkers for a poor response to vaccine treatment. Although further clarification is needed regarding the functions of miR-6826 and miR-6875 and their relationship to immune‑related molecules, plasma miR-6826 and miR-6875 may be useful negative biomarkers for predicting the efficacy of vaccine treatment.
Lung cancer is the most common cause of cancer deaths all over the world, in which non-small cell lung cancer (NSCLC) accounts for ~85% of cases. It is well known that microRNAs (miRNAs) play a critical role in various cellular processes, mediating post-transcriptional silencing either by mRNA degradation through binding the 3′ UTR of target mRNA or by translational inhibition of the protein. In the past decade, miRNAs have also been increasingly identified in biological fluids such as human serum or plasma known as circulating or cell-free miRNAs, and may function as non-invasive diagnostic markers for various cancer types including NSCLC. Circulating tumor cells (CTCs) are those cells that are shed from solid tumors and then migrate into the circulation. However, reports concerning the roles of CTCs are quite rare, which may be attributed to the difficulties in the enrichment and detection of CTCs in the circulation. Although, there have been reassuring advances in identifying circulating miRNA-panels, which are assumed to be of diagnostic value in NSCLC early stage, some issues remain concerning the reliability of using miRNA panels as a diagnostic tool for NSCLC. In the current review, we are aiming at providing insights into the miRNAs biology, the mechanisms of miRNAs release into the bloodstream, cell-free miRNAs as the diagnostic markers for NSCLC and the current limitations of CTCs as diagnostic markers in NSCLC.
Mammography is the predominant screening method for early detection of breast cancer, but has limitations and could be rendered more accurate by combination with a blood-based biomarker profile. Circulating microRNAs (miRNAs) are increasingly recognized as strong biomarkers, and we previously developed a 9-miRNA profile using serum and LNA-based qPCR that effectively stratified patients with early stage breast cancer vs. healthy women. To further develop the test into routine clinical practice, we collected serum of women examined by clinical mammography (N=197) according to standard operational procedures (SOPs) of the Danish Cancer Biobank. The performance of the circulating 9-miRNA profile was analyzed in 116 of these women, including 36 with breast cancer (aged 50-74), following a standardized protocol that mimicked a routine clinical set-up. We confirmed that the profile is significantly different between women with breast cancer and controls (p-value <0.0001), with an AUC of 0.61. Significantly, one woman whose 9-miRNA profile predicted a 73% probability of having breast cancer indeed developed the disease within one year despite being categorized as clinically healthy at the time of blood sample collection and mammography. We propose that this miRNA profile combined with mammography will increase the overall accuracy of early detection of breast cancer.
Prostate specific antigen (PSA) remains the most used biomarker in the management of early prostate cancer (PCa), in spite of the problems related to false positive results and overdiagnosis. New biomarkers have been proposed in recent years with the aim of increasing specificity and distinguishing aggressive from non-aggressive PCa. The emerging role of the prostate health index and the 4Kscore is reviewed in this article. Both are blood-based tests related to the aggressiveness of the tumor, which provide the risk of suffering PCa and avoiding negative biopsies. Furthermore, the use of urine has emerged as a non-invasive way to identify new biomarkers in recent years, including the PCA3 and TMPRSS2:ERG fusion gene. Available results about the PCA3 score showed its usefulness to decide the repetition of biopsy in patients with a previous negative result, although its relationship with the aggressiveness of the tumor is controversial. More recently, aberrant microRNA expression in PCa has been reported by different authors. Preliminary results suggest the utility of circulating and urinary microRNAs in the detection and prognosis of PCa. Although several of these new biomarkers have been recommended by different guidelines, large prospective and comparative studies are necessary to establish their value in PCa detection and prognosis.
Background
Recently, circulating microRNAs (miRNAs) have been reported to be stably detectable in plasma/serum and to function as potent biomarkers in various cancers. The aim of this study was to evaluate the expression level of plasma miRNA-195 in patients with non-small cell lung cancer (NSCLC) and investigate its diagnostic and prognostic value. Methods
Quantitative real-time PCR was performed to evaluate plasma miRNA-195 levels in 100 NSCLC patients and 100 healthy volunteers. The association between miRNA-195 expression and clinicopathological factors as well as the overall survival was analyzed. Receiver-operating characteristic (ROC) curve analysis was carried out to assess the potential value of plasma miRNA-195 for NSCLC diagnosis. ResultsPlasma miRNA-195 was downregulated in NSCLC patients compared with healthy controls (P < 0.001). Decreased plasma miRNA-195 expression was significantly associated with lymph node metastasis and advanced clinical stage. ROC curve analysis showed that plasma miRNA-195 was a useful marker for NSCLC diagnosis. Multivariate Cox regression analysis confirmed low plasma miRNA-195 expression as an independent unfavorable prognostic factor for NSCLC patients. Conclusions
These findings indicate that plasma miRNA-195 might serve as a promising biomarker for the early detection and prognosis evaluation of NSCLC.
Objective
Early diagnosis of epithelial ovarian cancer is critical for patient survival. The objective of this pilot study is to identify a circulating micro (mi)RNA as a potential biomarker for epithelial ovarian cancer.
Methods
A total of 135 epithelial ovarian cancer patients and 54 benign ovarian tumor patients were recruited for this study. Using customized TaqMan low density miRNA arrays, we first screened expression levels of 48 miRNAs in sera from 18 epithelial ovarian cancer patients and 16 benign ovarian tumor patients. The most significantly and differentially expressed miRNA was then further examined in all serum samples using real-time polymerase chain reaction. Its expression was further analyzed in relationship with clinicopathological factors and patient survival.
Results
Array screening data showed that expression levels of serum miRNA-20a, miRNA-125b, miRNA-126, miRNA-355, and let-7c were significantly different between malignant and benign ovarian tumor patients. Subsequent real-time polymerase chain reaction results showed that serum miRNA-125b levels were significantly higher in epithelial ovarian cancer patients compared to benign controls. Moreover, serum miRNA-125b levels were significantly higher in ovarian cancer patients in early stages I and II, and in patients having no residual tumor following surgery, but were not associated with differentiation and histological types of ovarian cancer. Notably, the higher level of miR-125b was significantly positively correlated with progression-free survival (P = 0.035) and marginally, with overall survival (P = 0.069).
Conclusions
miRNA-125b plays an important role in the pathogenesis and progression of epithelial ovarian cancer. Circulating miRNA-125b has the potential to become a novel biomarker for early diagnosis and prognosis prediction of epithelial ovarian cancer.
MicroRNAs (miRNAs) have been shown to be critical mediators of many cellular and developmental processes and have been implicated in different human diseases. Since the observation of extracellular miRNAs present in various biofluids, much attention and excitement have been garnered toward understanding the functional roles of these circulating extracellular miRNAs and establishing their potential use as noninvasive diagnostic biomarkers. Here, we will review the current state of miRNA biomarkers for many human diseases, including their emerging use in toxicological applications, and discuss the current challenges in the field, with an emphasis on technical issues that often hinder discovery-based miRNA biomarker studies.
Background:
Prostate cancer is the second most frequently diagnosed cancer in men worldwide. Theranostics, a combination of diagnostics and therapeutics, is an emerging concept in the field of precision medicine, and microRNAs (miRNAs) are predictive pioneers in this area.
Content:
miRNAs are small endogenous noncoding RNA molecules that regulate gene expression posttranscriptionally by targeting messenger RNAs. More than 60% of all protein coding genes are controlled by miRNAs, which makes them powerful regulators of the different cellular processes involved in the pathogenesis of various types of cancer, including prostate cancer. Growing evidence indicates the differential expression of miRNAs in tumor tissues. In addition, miRNAs in body fluids, known as circulating miRNAs, are present in remarkably stable forms and their alteration in prostate cancer has been well documented. Circulating miRNAs are known to originate from tumor tissues, thereby enabling intercellular communication via carriers to promote tumorigenesis and malignancy. In addition, fueled by recent advances, the use of miRNA-based anticancer therapies has been proposed with the onset of early phase clinical trials to assess the therapeutic efficacy of miRNAs.
Summary:
In this review, we summarize the theranostic utility of miRNAs and outline their diagnostic and prognostic potential in prostate cancer. In addition, we discuss the current detection methodologies and emerging innovative strategies for the detection of miRNAs in body fluids and tumor tissues in the clinical setting. We also provide insight into the current and future therapeutic potential of miRNAs in prostate cancer.
Objective:
To develop a plasma-based microRNA (miRNA) diagnostic assay specific for colorectal neoplasms, building upon our prior work.
Background:
Colorectal neoplasms [colorectal cancer (CRC) and colorectal advanced adenoma (CAA)] frequently develop in individuals at ages when other common cancers also occur. Current screening methods lack sensitivity, specificity, and have poor patient compliance.
Methods:
Plasma was screened for 380 miRNAs using microfluidic array technology from a "Training" cohort of 60 patients, (10 each) control, CRC, CAA, breast cancer, pancreatic cancer, and lung cancer. We identified uniquely dysregulated miRNAs specific for colorectal neoplasia (P < 0.05, false discovery rate: 5%, adjusted α = 0.0038). These miRNAs were evaluated using single assays in a "Test" cohort of 120 patients. A mathematical model was developed to predict blinded sample identity in a 150 patient "Validation" cohort using repeat-sub-sampling validation of the testing dataset with 1000 iterations each to assess model detection accuracy.
Results:
Seven miRNAs (miR-21, miR-29c, miR-122, miR-192, miR-346, miR-372, and miR-374a) were selected based upon P value, area under the curve (AUC), fold change, and biological plausibility. Area under the curve (±95% confidence interval) for "Test" cohort comparisons were 0.91 (0.85-0.96) between all neoplasia and controls, 0.79 (0.70-0.88) between colorectal neoplasia and other cancers, and 0.98 (0.96-1.0) between CRC and colorectal adenomas. In our "Validation" cohort, our mathematical model predicted blinded sample identity with 69% to 77% accuracy, 67% to 76% accuracy, and 86% to 90% accuracy for each comparison, respectively.
Conclusions:
Our plasma miRNA assay and prediction model differentiate colorectal neoplasia from patients with other neoplasms and from controls with higher sensitivity and specificity compared with current clinical standards.
There is an urgent need for novel noninvasive prognostic biomarkers for monitoring the recurrence of breast cancer. The purpose of this study is to identify circulating microRNAs that can predict breast cancer recurrence. We conducted a microRNA profiling experiment in serum samples from 48 breast cancer patients using Exiqon miRCURY microRNA RT-PCR panels. Significantly differentiated miRNAs for recurrence in the discovery profiling were further validated in an independent set of sera from 20 patients with breast cancer recurrences and 22 patients without recurrences. We identified seven miRNAs that were differentially expressed between breast cancer patients with and without recurrences, including four miRNAs upregulated (miR-21-5p, miR-375, miR-205-5p, and miR-194-5p) and three miRNAs downregulated (miR-382-5p, miR-376c-3p, and miR-411-5p) for recurrent patients. Using penalized logistic regression, we built a 7-miRNA signature for breast cancer recurrence, which had an excellent discriminating capacity (concordance index=0.914). This signature was significantly associated with recurrence after adjusting for known prognostic factors, and it was applicable to both hormone-receptor positive (concordance index=0.890) and triple-negative breast cancers (concordance index=0.942). We also found the 7-miRNA signature were reliably measured across different runs of PCR experiments (intra-class correlation coefficient=0.780) and the signature was significantly higher in breast cancer patients with recurrence than healthy controls (p=1.1x10-5). In conclusion, circulating miRNAs are promising biomarkers and the signature may be developed into a minimally invasive multi-marker blood test for continuously monitoring the recurrence of breast cancer. It should be further validated for different subtypes of breast cancers in longitudinal studies.
Ovarian cancer is the leading cause of death among gynecologic malignancies. Since ovarian cancer develops asymptomatically, it is often diagnosed at an advanced and incurable stage. Despite many years of research, there is still a lack of reliable diagnostic markers and methods for early detection and screening. Recently, it was discovered that cell-free microRNAs (miRNAs) circulate in the body fluids of healthy and diseased patients, suggesting that they may serve as a novel diagnostic marker. This review summarizes the current knowledge regarding the potential clinical relevance of circulating cell-free miRNA for ovarian cancer diagnosis, prognosis, and therapeutics. Despite the high levels of ribonucleases in many types of body fluids, most of the circulating miRNAs are packaged in microvesicles, exosomes, or apoptotic bodies, are binding to RNA-binding protein such as argonaute 2 or lipoprotein complexes, and are thus highly stable. Cell-free miRNA signatures are known to be parallel to those from the originating tumor cells, indicating that circulating miRNA profiles accurately reflect the tumor profiles. Since it is well established that the dysregulation of miRNAs is involved in the tumorigenesis of ovarian cancer, cell-free miRNAs circulating in body fluids such as serum, plasma, whole blood, and urine may reflect not only the existence of ovarian cancer but also tumor histology, stage, and prognoses of the patients. Several groups have successfully demonstrated that serum or plasma miRNAs are able to discriminate patients with ovarian cancer patients from healthy controls, suggesting that the addition of these miRNAs to current testing regimens may improve diagnosis accuracies for ovarian cancer. Furthermore, recent studies have revealed that changes in levels of cell-free circulating miRNAs are associated with the condition of cancer patients. Discrepancies between the results across studies due to the lack of an established endogenous miRNA control to normalize for circulating miRNA levels, as well as differing extraction and quantification methods, are the pitfalls to be resolved before clinical application. There is still a long way, however, before this can be achieved, and further evidence would make it possible to apply circulating cell-free miRNAs not only as biomarkers but also as potential therapeutic targets for ovarian cancer in the future.
Improving early detection of lung cancer is critical to improving lung cancer survival. Studies have shown that computerized tomography (CT) screening can reduce mortality from lung cancer, but this involves risks of radiation exposure and can identify non-cancer lung nodules that lead to unnecessary interventions for some. There is a critical need to develop alternative, less invasive methods to identify patients who have early-stage lung cancer. The detection of circulating tumor cells (CTCs) are a promising area of research, but current technology is limited by a low yield of CTCs. Alternate studies are investigating circulating nucleic acids and proteins as possible tumor markers. It is critical to develop innovative methods for early lung cancer detection that may include CTCs or other markers that are low-risk and low-cost, yet specific and sensitive, to facilitate improved survival by diagnosing the disease when it is surgically curable.
Background:
Prostate cancer (PCa) screening currently relies on prostate-specific antigen (PSA) testing and digital rectal examination. However, recent large-scale studies have questioned the long-term efficacy of these tests, and biomarkers that accurately identify PCa are needed.
Methods:
We analysed the levels of circulating microRNAs (miRNAs) in patients with elevated PSA who were diagnosed with either localised PCa (n=36) or benign prostatic hyperplasia (BPH, n=31) upon biopsy. Real-time RT-PCR with Taqman probes was used to measure plasma levels of miRNAs. To circumvent problems associated with circulating miRNA quantitation, we computed the expression ratios of upregulated and downregulated miRNAs.
Results:
The miR-106a/miR-130b and miR-106a/miR-223 ratios were significantly different between the biopsy-positive and BPH groups (P<0.0001), and yielded statistical power values that were >0.99. Both miRNA ratios were highly sensitive and more specific than PSA in discriminating localised PCa from BPH. Receiver operating characteristic curve analysis revealed area under curve values of 0.81 (miR-106a/miR-130b) and 0.77 (miR-106a/miR-223).
Conclusions:
Testing for circulating miR-106a/miR-130b and miR-106a/miR-223 ratios may reduce the costs and morbidity of unnecessary biopsies and is feasible for large-scale screening, as it requires measuring only three miRNAs.British Journal of Cancer advance online publication, 26 May 2016; doi:10.1038/bjc.2016.151 www.bjcancer.com.
The effective and efficient management of cancer patients relies upon early diagnosis and/or the monitoring of treatment, something that is often difficult to achieve using standard tissue biopsy techniques. Biological fluids such as blood hold great possibilities as a source of non-invasive cancer biomarkers that can act as surrogate markers to biopsy-based sampling. The non-invasive nature of these "liquid biopsies" ultimately means that cancer detection may be earlier and that the ability to monitor disease progression and/or treatment response represents a paradigm shift in the treatment of cancer patients. Below, we review one of the most promising classes of circulating cancer biomarkers: microRNAs (miRNAs). In particular, we will consider their history, the controversy surrounding their origin and biology, and, most importantly, the hurdles that remain to be overcome if they are really to become part of future clinical practice.
The ability of the classic tumour-suppressive let-7 family to inhibit carcinogenesis, tumour progression, recurrence and pluripotency of cancer stem cells has generated significant interest in the field of cancer research. Through suppressing and degrading downstream-targeted mRNAs, let-7 affected most aspects of cell biology. It is perplexing how let-7 affects oncogenesis, as the large influx of new miRNAs and other kinds of non-coding RNAs are continuously defined. In this review, we delineate the complex functions of let-7 and discuss the future direction of let-7 research.
Background:
The diagnostic value of circulating microRNAs (miRNAs) detection in prostate cancer (PC) patients is currently under debate. Thus, we performed this meta-analysis of published literature to systematically evaluate the diagnostic potential of circulating miRNAs in PC.
Methods:
Eligible studies were searched in PubMed, Embase and Chinese National Knowledge Infrastructure databases. Sensitivity and specificity were pooled using a random-effects model and were used to plot the summary receiver operator characteristic (SROC) curve. All analyses were performed using the Stata 13.0 software and Meta-Disc 1.4 for windows.
Results:
A total of ten articles were included for the meta-analysis according to the inclusion criteria. The pooled results based on all included studies showed circulating miRNAs have a relatively good diagnostic performance, with a sensitivity of 0.74, a specificity of 0.71 and an area under SROC curve (AUC) of 0.77 in indiscriminating PC from controls. Furthermore, meta-regression and subgroup analyses indicate that multiple circulating miRNAs detection displayed a better diagnostic performance than single one, with an AUC rising from 0.75 to 0.81. Additional interesting finding was that Caucasian-based circulating miRNAs assays could reach a higher accuracy compared with non-Caucasian-based one for PC with the p value of 0.0378.
Conclusion:
Our results confirmed the potential use of circulating miRNAs in the early diagnosis of PC, especially the combination of multiple circulating miRNAs. However, large-scale prospective studies are still needed to further validate our findings.
Background: Recent evidence suggests that microRNAs play an important role in cancer diagnostics. We assessed plasma microRNA-21 levels in patients with colorectal cancer (CRC) at different stages and in patients with benign polyps.
Methods: Plasma levels of miR-21 were assessed by quantitative reverse transcription polymerase chain reaction assay in plasma samples of 76 CRC patients and in 20 patients with benign polyps. Differences between groups were evaluated with Mann-Whitney and Kruskal-Wallis tests.
Results: No significant differences of miR-21 plasma levels were observed between CRC patients and subjects with benign polyps (p > 0.05). Also, no significant differences were found between CRC patients with advanced (III-IV) or early cancer stages (I-II) (p > 0.05).
Conclusions: These results do not support the hypothesis that circulating miR-21 expression is increased in adenoma-carcinoma-advanced carcinoma sequence. Accordingly, plasma miR-21 assessment does not appear to be a useful biomarker for diagnosing and staging CRC.
Lung cancer is the most common cancer in the world and the leading cause of tumor death among males. MicroRNAs (miRNAs) are single-stranded RNAs of approximately 22 nucleotides and constituted a new class of gene regulators in humans. As a novel class of emerging biomarkers, the aberrant expression of miRNA has been detected in various tumors. miRNAs are secreted into circulation by microvesicles from the broken tumor cells and act as either oncogenes or tumor suppressors in tumor tissues. In this review, we summarized different circulating miRNAs and their expression level as well as predictable values in lung cancer patients which were investigated in recent 5 years. Circulating miRNAs are found to be dysregulated and have association with clinicopathological parameters and overall survival in lung cancer patients. In conclusion, circulating miRNAs have the potential for distinguishing lung cancer patients from healthy individuals, with the advantages of stabilities, noninvasiveness and cost-effectiveness.
In this chapter, we outline various traditions of thought and explain how Mediated Discourse Analysis borrows insights and analytical tools from many of them. We then move on and illustrate that the framework proposed, however differs from all of the other approaches in a number of important ways: The difference, we argue can perhaps be summed up best by transforming the slogan discourse as action into discourse in action.
Exosomes are membrane vesicles that mediate intercellular communication by transporting their molecular cargo from cell to cell. We investigated whether serum levels of exosomal miR-373, miR-200a, miR-200b and miR-200c and circulating exosomes have diagnostic and prognostic relevance in a cohort of 163 epithelial ovarian cancer (EOC) patients using TaqMan MicroRNA assays and ELISA. The serum concentrations of exosomal miR-373 (p = 0.0001), miR-200a (p = 0.0001), miR-200b (p = 0.0001) and miR-200c (p = 0.028) were significantly higher in EOC patients than healthy women. The levels of miR-200a (p = 0.0001), miR-200b (p = 0.0001) and miR-200c (p = 0.019) could distinguish between malignant and benign ovarian tumors. While the levels of miR-373 and miR-200a were increased in all FIGO/lymph node stages (p = 0.0001), the levels of miR-200b and miR-200c were higher in patients with FIGO stage III-IV (p = 0.0001, p = 0.008, respectively) including lymph node metastasis (p = 0.0001, p = 0.004, respectively) than FIGO stages I-II. The increased levels of miR-200b and miR-200c were also associated with CA125 values (p = 0.0001, p = 0.0001, respectively) and a shorter overall survival (p = 0.007, p = 0.017, respectively). The levels of exosomes were excessively elevated in EOC patients (p = 0.0001). In all three cohorts, they were positively associated with the serum levels of exosomal miR-373 (p = 0.004), miR-200a (p = 0.0001), miR-200b (p = 0.0001) and miR-200c (p = 0.008). In conclusion, the increased levels of exosomal miR-200b and miR-200c mainly observed in advanced EOC suggest that these microRNAs may be involved in tumor progression. The high concentrations of exosomes in EOC patients imply an excessive, active exosomal secretion in EOC.
Exosomes have been shown to transmit drug resistance through delivering miRNAs. We aimed to explore their roles in breast cancer. Three resistant sublines were established by exposing parental MDA-MB-231 cell line to docetaxel, epirubicin and vinorelbine, respectively. Preneoadjuvant chemotherapy biopsies and paired surgically-resected specimens embedded in paraffin from 23 breast cancer patients were collected. MiRNA expression profiles of the cell lines and their exosomes were evaluated using microarray. The result showed that most miRNAs in exosomes had a lower expression level than that in cells, however, some miRNAs expressed higher in exosomes than in cells, suggesting a number of miRNAs is concentrated in exosomes. Among the dysregulated miRNAs, 22 miRNAs were consistently up-regulated in exosomes and their cells of origin. We further found that 12 of the 22 miRNAs were significantly up-regulated after preneoadjuvant chemotherapy. Further study of the role of these 12 miRNAs in acquisition of drug resistance is needed to clarify their contribution to chemoresistance.
Growing evidence is showing that metastatic cell populations are able to transfer their characteristics to less malignant cells. Exosomes (EXOs) are membrane vesicles of endocytic origin able to convey their cargo of mRNAs, microRNAs (miRs), proteins and lipids from donors to proximal as well as distant acceptor cells. Our previous results indicated that miR-221&222 are key factors for melanoma development and dissemination. The aim of this study was to verify whether the tumorigenic properties associated with miR-222 overexpression can be also propagated by miR-222-containing EXOs.
EXOs were isolated by UltraCentrifugation or Exoquick-TC ® methods. Preparations of melanoma-derived vesicles were characterized by using the Nanosight™ technology and the expression of exosome markers analyzed by western blot. The expression levels of endogenous and exosomal miRNAs were examined by real time PCR. Confocal microscopy was used to evaluate transfer and uptake of microvesicles from donor to recipient cells. The functional significance of exosomal miR-222 was estimated by analyzing the vessel-like process formation, as well as cell cycle rates, invasive and chemotactic capabilities.
Besides microvesicle marker characterization, we evidenced that miR-222 exosomal expression mostly reflected its abundance in the cells of origin, correctly paralleled by repression of its target genes, such as p27Kip1, and induction of the PI3K/AKT pathway, thus confirming its functional implication in cancer. The possible differential significance of PI3K/AKT blockade was assessed by using the BKM120 inhibitor in miR-222-transduced cell lines. In addition, in vitro cultures showed that vesicles released by miR-222-overexpressing cells were able to transfer miR-222-dependent malignancy when taken-up by recipient primary melanomas. Results were confirmed by antagomiR-221&222 treatments and by functional observations after internalization of EXOs devoid of these miRs.
All together these data, besides generally confirming the role of miR-222 in melanoma tumorigenesis, supported its responsibility in the exosome-associated melanoma properties, thus further indicating this miR as potential diagnostic and prognostic biomarker and its abrogation as a future therapeutic option.
One of the most fundamental issues in discourse studies is understanding the relationship between what we say and write and what we do, that is, the relationship between discourse and action. Nearly all mainstream approaches to discourse analysis begin with the assumption that discourse itself is a kind of social action. Conversation analysts, for example, see talk as a matter of “paired action,” genre analysts speak of genres as examples of “communicative actions,” interactional sociolinguists focus on how people strategize discursive actions within an ongoing negotiative process with other social actors, and critical discourse analysts remind us that discourse constitutes and is constitutive of what they call “social practice.”
Keywords:
discourse analysis;
language and social interaction;
research methods in applied linguistics
The Hippo signalling pathway co-ordinately modulates cell regeneration and organ size, and its deregulation contributes to tumourigenesis through many cellular processes, including over-proliferation, apoptosis resistance, and cell migration. Recent discoveries have shed new light on how microRNAs (miRNAs) are closely linked to the Hippo pathway in tumour progression. Hippo signalling has been reported to affect widespread miRNA biogenesis. In turn, several miRNAs regulate Hippo signalling, which contributes to carcinogenesis. This article will provide an overview of the crosstalk between Hippo signalling and miRNAs in the development of cancer and further appraise potential targets for therapeutic intervention. This article is protected by copyright. All rights reserved.
Prostate cancer (CaP) is a leading cause of cancer death and displays a broad range of clinical behavior from relatively indolent to aggressive metastatic disease. Due to the alteration and incomplete characterization of the CaP genomic markers, the quest for novel cellular metabolic regulatory molecules like micro RNA (miRNA) as a biomarker could be considered for the prognosis and treatment of CaP in future. In this article, we review the existing literature pertaining to CaP. Study provides a comprehensive miRNA profile expressed in CaP. Beside the miRNA expressed in the tumor tissue, circulating miRNAs have been found highly stable and are both detectable and quantifiable in a range of accessible bio fluids; therefore, miRNA has the potential to be useful diagnostic, prognostic and predictive biomarker. Along with being an important molecule in modulation of CaP progression, the miRNA have certain limitations such as lack of stable expression of multiple target genes and often disrupt entire signaling networks of cellular metabolic pathways. We conclude that: The alteration of miRNA and their role played in cellular regulatory networks would be the next target of basic research in CaP. The miRNAs identified may be validated and modeled to understand their role in CaP, using bioinformatics. There is an immediate unmet need in the translational approach of identified miRNAs. The characterization of miRNAs involved in CaP is still incomplete: adequate validation studies are required to corroborate current results.
MicroRNAs (miRNAs) are short, non‑coding RNA molecules that act as regulators of gene expression. Circulating blood miRNAs have potential as cancer biomarkers. The main objective of the present study was to assess the effect of miRNA‑23b (miR‑23b) expression in plasma on the diagnosis and prognosis of colorectal cancer (CRC). Reverse transcription‑quantitative polymerase chain reaction (PCR) was used to measure miR‑23b expression levels, and methylation‑specific PCR was used to test the promoter methylation status. Subsequently, the expression level of miR‑23b in plasma samples was compared between CRC patients and healthy control individuals. The miR‑23b expression levels were significantly lower in CRC cells and primary CRC tissues than in nonmalignant colorectal tissues (P<0.001). It was also shown that miR‑23b expression is downregulated by promoter methylation and can be restored by demethylation agent treatment. miR‑23b was significantly decreased in plasma samples from CRC patients compared with the healthy control individuals (P<0.001). The value of the area under the receiver operating characteristic curve was 0.842 (sensitivity, 84.38%; specificity, 77.08%; 95% confidence interval, 0.763‑0.922). Low plasma miR‑23b expression was significantly associated with clinical stage, tumor depth, distant metastasis and tumor recurrence. CRC patients with low miR‑23b expression in plasma exhibited a shorter recurrence‑free survival time and poorer overall survival rate. The present results suggested that the downregulation of miR‑23b in the plasma has the potential to be a diagnostic and prognostic biomarker in CRC.
Background
Circulating microRNAs (miRNAs) are attracting major interest as potential noninvasive biomarkers for colorectal cancer (CRC). This study aimed to identify a novel serum miRNA biomarker for the early detection and/or evaluating prognosis of CRC patients.
Patients and methods
Comprehensive miRNA array analysis was performed using serum samples from patients with colorectal neoplasia and healthy controls. Next, to verify whether the candidate miRNA possessed a secretory potential, we screened miRNA expression levels in culture medium from 2 CRC cell lines, followed by serum analysis from 12 stage IV CRC, 12 adenoma and 12 control subjects. Thereafter, we validated expression of candidate miRNAs in 179 primary CRC tissues, as well as serum samples from an independent cohort of 211 CRCs, 56 adenomas, and 57 control subjects.
Results
Through microarray analysis, we identified significantly higher levels of miRNA-1290 (miR-1290) in serum from patients with colorectal adenomas and cancers. We verified miR-1290 overexpression in serum of CRC patients in a training cohort. In the validation cohort, serum miR-1290 levels were significantly upregulated in patients with colorectal adenomas (P<0.0001) and cancers (P<0.0001). Serum miR-1290 levels could robustly distinguish adenoma (area under the curve [AUC]=0.718) and CRC patients (AUC=0.830) from normal subjects. High miR-1290 expression in serum and tissue was significantly associated with tumor aggressiveness and poor prognosis. Moreover, serum miR-1290 levels were an independent prognostic factor (hazard ratio=4.51; 95% confidence interval=1.23-23.69; P=0.0096) and an independent predictor for tumor recurrence (hazard ratio=3.92; 95% confidence interval=1.11-25.14; P=0.032) in CRC.
Conclusions
Serum miR-1290 is a novel biomarker for early detection, recurrence and prognosis in CRC.
The contribution of microRNAs to the regulation of mRNA expression during physiological and developmental processes are well-recognized. These roles are being expanded by recent observations that emphasize the capability of miRNA to participate in inter-cellular signaling and communication. Several factors support a functional role for miRNA as mediators of cell-to-cell signaling. miRNA are able to exist within the extracellular milieu or circulation, and their stability and integrity maintained through association with binding proteins or lipoproteins, or through encapsulation within cell-derived membrane vesicles. Furthermore, miRNA can retain functionality and regulate target gene expression following their uptake by recipient cells. In this overview, we review specific examples that will highlight the potential of miRNA to serve as paracrine signaling mediators in metabolic diseases and cancers. Elucidating the mechanisms involved in inter-cellular communication involving miRNA will provide new insights into disease pathogenesis and potential therapeutic opportunities.
Early detection of colorectal cancer is the main prerequisite for successful treatment and reduction of mortality. Circulating microRNAs were previously identified as promising diagnostic, prognostic and predictive biomarkers. The purpose of the present study was to identify serum microRNAs enabling early diagnosis and prognosis prediction of colon cancer. In total, serum samples from 427 colon cancer patients and 276 healthy donors were included in 3-phase biomarker study. Large-scale microRNA expression profiling was performed using Illumina small RNA sequencing. Diagnostic and prognostic potential of identified microRNAs was validated on independent training and validation sets of samples using RT-qPCR. Fifty-four microRNAs were found to be significantly deregulated in serum of colon cancer patients compared to healthy donors (P < 0.01). A diagnostic 4-microRNA signature consisting of miR-23a-3p, miR-27a-3p, miR-142-5p and miR-376c-3p was established (AUC = 0.917), distinguishing colon cancer patients from healthy donors with sensitivity of 89% and specificity of 81% (AUC = 0.922). This panel of microRNAs exhibited high diagnostic performance also when analyzed separately in colon cancer patients in early stages of the disease (T1-4N0M0; AUC = 0.877). Further, a prognostic panel based on the expression of miR-23a-3p and miR-376c-3p independent of TNM stage was established (HR 2.30; 95% CI 1.44-3.66; P < 0.0004). In summary, highly sensitive signatures of circulating microRNAs enabling non-invasive early detection and prognosis prediction of colon cancer were identified.
The assessment of differentially expressed microRNAs in patients and healthy controls is important to identify potential tumor biomarkers. Recently, it has been shown that the microRNA levels in exosomes are more correlated with the clinical-pathological variables than vesicle-free microRNAs (miRNAs) in biofluids; therefore, there is an increasing interest in these specific evaluations. However, these measurements can be affected by experimental problems that not always are evaluated and/or by inadequate procedural choices. In particular, exosome isolation and miRNA extraction procedures are crucial to avoid contaminations, and even the choice of the most suitable purity controls is important. Moreover, a stable endogenous RNA should be used for normalization of miRNA expression obtained by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) in order to make these measures comparable among different samples. A rushed choice of the endogenous control can bias study conclusions without revealing inconsistencies. Unfortunately, a few studies systematically identified the best normalizer for their specific experimental context. Instead, sometimes, the normalization procedures were performed in a disputable way or the normalizer choices simply based on the previous literature. Here, we reviewed the studies where the exosomal miRNA profiling was assessed in human biofluids to point out the adopted procedures and the specific endogenous controls chosen for normalization.
MicroRNAs (miRNA) are short, noncoding RNAs whose dysregulation has been implicated in most, if not all, cancers. They regulate gene expression by suppressing mRNA translation and reducing mRNA stability. To this end, there is a great deal of interest in modifying miRNA expression levels for the treatment of cancer. However, the literature is fraught with inconsistent accounts as to whether various miRNAs are oncogenic or tumor suppressive. In this review, we directly examine these inconsistencies and propose several mechanisms to explain them. These mechanisms include the possibility that specific miRNAs can simultaneously produce competing oncogenic and tumor suppressive effects by suppressing both tumor suppressive mRNAs and oncogenic mRNAs, respectively. In addition, miRNAs can modulate tumor-modifying extrinsic factors, such as cancer-immune system interactions, stromal cell interactions, oncoviruses, and sensitivity to therapy. Ultimately, it is the balance between these processes that determines whether a specific miRNA produces a net oncogenic or net tumor suppressive effect. A solid understanding of this phenomenon will likely prove valuable in evaluating miRNA targets for cancer therapy. Cancer Res; 76(13); 1-5. ©2016 AACR.
Despite recent advances, non-small cell lung cancer remains a devastating disease and carries a grim prognosis. Major contributing factors include difficulties in diagnosing the disease early in its course during the asymptomatic stage and the poor understanding of the biology underlying disease progression. Liquid biopsies, noninvasive blood tests that detect circulating biomarkers such as circulating tumor cells and tumor-derived nucleic acid fragments, are a rapidly evolving field of research that could provide answers to both of these unmet needs. Herein, we review the relevant data concerning the diagnostic, predictive and prognostic significance of three distinct but potentially complementary circulating biomarkers in non-small cell lung cancer: circulating tumor cells, cell free DNA and microRNAs.
The discovery of cell-free microRNAs (miRNAs) in serum, plasma and other body fluids has yielded an invaluable potential source of non-invasive biomarkers for cancer and other non-malignant diseases. miRNAs in the blood and other body fluids are highly stable in biological samples and are resistant to environmental conditions, such as freezing, thawing or enzymatic degradation, which makes them convenient as potential biomarkers. In addition, they are more easily sampled than tissue miRNAs. Altered levels of cell-free miRNAs have been found in every type of cancer analysed, and increasing evidence indicates that they may participate in carcinogenesis by acting as cell-to-cell signalling molecules. This review summarizes the biological characteristics and mechanisms of release of cell-free miRNAs that make them promising candidates as non-invasive biomarkers of cancer.
Multimodal discourse analysis is an emergent field that began around the verge of the millennium with books such as Reading Images: A Grammar of Visual Design (Kress & van Leeuwen, 1996/2006), Mediated Discourse as Social Interaction (Scollon, 1998), Multimodal Discourse (Kress & van Leeuwen, 2001), and Analyzing Multimodal Interaction: A Methodological Framework (Norris, 2004). Initially, multimodal discourse analysis was primarily the domain of mediated discourse analysts, social semioticians, and systemic functional linguists. While early developments were somewhat overlapping in time, these works resulted from, and aligned with, two separate major paradigm shifts stemming from previous work in discourse analysis...
Objective:
The purpose of this study was to explore serum miR-135a-5p expression in colorectal cancer and examine the potential usefulness of this molecule as a biomarker for diagnosis in colorectal cancer.
Methods:
Serum samples were collected from 60 patients with primary colorectal cancer, 40 patients with colorectal polyps and 50 healthy controls. Serum miR-135a-5p expression levels were detected by reverse transcription quantitative real-time quantitative polymerase chain reaction. Serum carcinoembryonic antigen and carbohydrate antigen 199 concentrations were detected by MODULAR ANALYTICS E170.
Results:
The relative expression level of serum miR-135a-5p in colorectal cancer patients, colorectal polyps patients and healthy controls was 2.451 (1.107, 4.413), 0.946 (0.401, 1.942) and 0.949 (0.194, 1.415), respectively, indicating that it was significantly higher in colorectal cancer patients than that in the other two groups (U = 351.0, 313.0, both P < 0.001). Additionally, it was significantly correlated with different degrees of tumour differentiation (U = 215.0, P = 0.029) and different tumour stages (U = 202.0, P = 0.013). There was no significant correlation between the relative expression of serum miR-135a-5p and carcinoembryonic antigen (r(2 )= 0.023, P = 0.293) or carbohydrate antigen 199 (r(2 )= 0.067, P = 0.068) in colorectal cancer patients. Compared with colorectal polyps group, AUC(ROC) of serum miR-135a-5p in colorectal cancer group was 0.832 with 95% CI 0.73-0.93; compared with healthy control group, AUC(ROC) was 0.875 with 95% CI 0.80-0.95.
Conclusion:
Serum miR-135a-5p expression in colorectal cancer patients was higher than that in patients with colorectal polyps and healthy controls, suggesting that serum miR-135a-5p may prove to be an important biomarker for auxiliary diagnosis of colorectal cancer.
Moving towards multimodal mediated theory, I propose to define a mode as a system of mediated action that comes about through concrete lower-level actions that social actors take in the world. In order to explain exactly how a mode is a system of mediated action, I turn to a perfume blog and use one blog entry as my starting point. The mode that I primarily focus on in this article is the mode of smell, explicating that the mode of smell is not synonymous with olfactory perception, even though modal development of smell is certainly partially dependent upon olfactory perception.
As I am ostensibly focusing on the one mode, I once again problematize this notion of countability and delineate the purely theoretical and heuristic unit of mode (Norris, 2004). I clarify that modes a) do not exist in the world as they are purely theoretical in nature; b) that modes can be delineated in various ways; and c) that modes are never singular.
Even though the concept of mode is problematical – and in my view needs to always be problematized – I argue that the term and the notion of mode is theoretically useful as it allows us to talk about and better understand communication and (inter)action in three respects: 1. The notion of mode allows us to investigate regularities as residing on a continuum somewhere between the social actor(s) and the mediational means; 2. The theoretical notion of mode embraces socio-cultural and historical as well as individual characteristics, never prioritising any of these and always embracing the tension that exists between social actor(s) and mediational means; and 3. The theoretical notion of mode demonstrates that modal development through concrete lower-level actions taken in the world, is transferable to other lower-level actions taken.
Divorce is a traumatic disruption in the lives of families that puts both parents and children at risk for long-term emotional and social consequences. However, if the non-residential parent maintains a quality relationship with the child, many of these negative consequences are mitigated. Divorced families face substantial challenges in parenting while living apart, especially as geographic separation often makes in-person visitation more difficult. Many families are turning to virtual visitation—supplementing in-person visits with use of communication technologies such as videoconferencing. However, current communication technologies are often inadequate to support long-distance parenting. We discuss the needs of divorced families and how these may be addressed through design. We present a case study of a single intervention, called the ShareTable, aimed at enriching virtual visitation between parents and children who live apart. Finally, we discuss the challenges and opportunities of designing for divorced families.