Single-cell multi-omics of human clonal hematopoiesis reveals that DNMT3A R882 mutations perturb early progenitor states through selective hypomethylation

Somatic mutations in cancer genes have been ubiquitously detected in clonal expansions across healthy human tissue, including in clonal hematopoiesis. However, mutated and wildtype cells are morphologically and phenotypically similar, limiting the ability to link genotypes with cellular phenotypes. To overcome this limitation, we leveraged multi-modality single-cell sequencing, capturing the mutation with transcriptomes and methylomes in stem and progenitors from individuals with DNMT3A R882 mutated clonal hematopoiesis. DNMT3A mutations resulted in myeloid over lymphoid bias, and in expansion of immature myeloid progenitors primed toward megakaryocytic-erythroid fate. We observed dysregulated expression of lineage and leukemia stem cell markers. DNMT3A R882 led to preferential hypomethylation of polycomb repressive complex 2 targets and a specific sequence motif. Notably, the hypomethylation motif is enriched in binding motifs of key hematopoietic transcription factors, serving as a potential mechanistic link between DNMT3A R882 mutations and aberrant transcriptional phenotypes. Thus, single-cell multi-omics pave the road to defining the downstream consequences of mutations that drive human clonal mosaicism.

Low guanine content and biased nucleotide distribution in vertebrate mtDNA can cause overestimation of non-CpG methylation

ABSTRACT Mitochondrial DNA (mtDNA) methylation in vertebrates has been hotly debated for over 40 years. Most contrasting results have been reported following bisulfite sequencing (BS-seq) analyses. We addressed whether BS-seq experimental and analysis conditions influenced the estimation of the levels of methylation in specific mtDNA sequences. We found false positive non-CpG methylation in the CHH context (fpCHH) using unmethylated Sus scrofa mtDNA BS-seq data. fpCHH methylation was detected on the top/plus strand of mtDNA within low guanine content regions. These top/plus strand sequences of fpCHH regions would become extremely AT-rich sequences after BS-conversion, whilst bottom/minus strand sequences remained almost unchanged. These unique sequences caused BS-seq aligners to falsely assign the origin of each strand in fpCHH regions, resulting in false methylation calls. fpCHH methylation detection was enhanced by short sequence reads, short library inserts, skewed top/bottom read ratios and non-directional read mapping modes. We confirmed no detectable CHH methylation in fpCHH regions by BS-amplicon sequencing. The fpCHH peaks were located in the D-loop, ATP6, ND2, ND4L, ND5 and ND6 regions and identified in our S. scrofa ovary and oocyte data and human BS-seq data sets. We conclude that non-CpG methylation could potentially be overestimated in specific sequence regions by BS-seq analysis.

Experiential design on cause-related marketing: lessons from the fill-the-bottle challenge

Purpose Based on a “Fill-the-Bottle” (FTB) challenge, this research explores how experiential design can help cause-related marketing. This study aims to show that experiences designed as anti-structural and anti-functional can raise awareness through action. Design/methodology/approach The authors study a corpus of 52 introspective journals and 60 pictures about the challenge, which entails filling empty bottles with cigarette butts from the streets as quickly as possible, then sharing pictures of the bottles on social media. Findings The anti-structural design of the experience activates the participants’ experiential system, and the social interactions between the participants and pedestrians construct meaning for the experience. The results further indicate that as follows: individuals’ frames of reference can explain whether they perceive the experience as liberatory or stochastic; anti-structural design can serve cause-related marketing by focusing on three stages: doing, showing and sharing; and experiential marketing can serve societal and social causes. Research limitations/implications This research involved a single field. Further research with more heterogeneous participants would be insightful. The power of experiential marketing to serve meaningful and collective causes should be encouraged. Further research should be conducted to understand and conceptualize these collective attempts to fight the dark sides of consumption. Practical implications In line with Pine and Gilmore’s (1999) advice to stage memorable experiences by working cautiously on cues, the FTB challenge analysis indicates that by focusing on material evidence and staging a specific sequence of doing something about it, showing everyone what is being done and expanding visibility by sharing artifacts of the action on social media, one can actually make people think about and remember the action. Social implications The “do-show-share” design that the FTB challenge uses can be relevant for many cause-related marketing efforts because it operates on both individual and collective levels. Originality/value This research offers a new perspective on experiential marketing by studying how experiences designed to be anti-structural can renew social, cause-related marketing tools.

Magnetic Resonance Imaging Images under Deep Learning in the Identification of Tuberculosis and Pneumonia

This work aimed to explore the application value of deep learning-based magnetic resonance imaging (MRI) images in the identification of tuberculosis and pneumonia, in order to provide a certain reference basis for clinical identification. In this study, 30 pulmonary tuberculosis patients and 27 pneumonia patients who were hospitalized were selected as the research objects, and they were divided into a pulmonary tuberculosis group and a pneumonia group. MRI examination based on noise reduction algorithms was used to observe and compare the signal-to-noise ratio (SNR) and carrier-to-noise ratio (CNR) of the images. In addition, the apparent diffusion coefficient (ADC) value for the diagnosis efficiency of lung parenchymal lesions was analyzed, and the best b value was selected. The results showed that the MRI image after denoising by the deep convolutional neural network (DCNN) algorithm was clearer, the edges of the lung tissue were regular, the inflammation signal was higher, and the SNR and CNR were better than before, which were 119.79 versus 83.43 and 12.59 versus 7.21, respectively. The accuracy of MRI based on a deep learning algorithm in the diagnosis of pulmonary tuberculosis and pneumonia was significantly improved (96.67% vs. 70%, 100% vs. 62.96%) ( P < 0.05 ). With the increase in b value, the CNR and SNR of MRI images all showed a downward trend ( P < 0.05 ). Therefore, it was found that the shadow of tuberculosis lesions under a specific sequence was higher than that of pneumonia in the process of identifying tuberculosis and pneumonia, which reflected the importance of deep learning MRI images in the differential diagnosis of tuberculosis and pneumonia, thereby providing reference basis for clinical follow-up diagnosis and treatment.

Discovery of an ancient MHC category with both class I and class II features

Two classes of major histocompatibility complex (MHC) molecules, MHC class I and class II, play important roles in our immune system, presenting antigens to functionally distinct T lymphocyte populations. However, the origin of this essential MHC class divergence is poorly understood. Here, we discovered a category of MHC molecules (W-category) in the most primitive jawed vertebrates, cartilaginous fish, and also in bony fish and tetrapods. W-category, surprisingly, possesses class II–type α- and β-chain organization together with class I–specific sequence motifs for interdomain binding, and the W-category α2 domain shows unprecedented, phylogenetic similarity with β2-microglobulin of class I. Based on the results, we propose a model in which the ancestral MHC class I molecule evolved from class II–type W-category. The discovery of the ancient MHC group, W-category, sheds a light on the long-standing critical question of the MHC class divergence and suggests that class II type came first.

Communication pathways bridge local and global conformations in an IgG4 antibody

The affinity of an antibody for its antigen is primarily determined by the specific sequence and structural arrangement of the complementarity-determining regions (CDRs). Recent evidence, however, points toward a nontrivial relation between the CDR and distal sites: variations in the binding strengths have been observed upon mutating residues separated from the paratope by several nanometers, thus suggesting the existence of a communication network within antibodies, whose extension and relevance might be deeper than insofar expected. In this work, we test this hypothesis by means of molecular dynamics (MD) simulations of the IgG4 monoclonal antibody pembrolizumab, an approved drug that targets the programmed cell death protein 1 (PD-1). The molecule is simulated in both the apo and holo states, totalling 4 μs of MD trajectory. The analysis of these simulations shows that the bound antibody explores a restricted range of conformations with respect to the apo one, and that the global conformation of the molecule correlates with that of the CDR. These results support the hypothesis that pembrolizumab featues a multi-scale hierarchy of intertwined global and local conformational changes. The analysis pipeline developed in this work is general, and it can help shed further light on the mechanistic aspects of antibody function.

Emotion Modulation through Music after Sadness Induction—The Iso Principle in a Controlled Experimental Study

Music therapy intervention manuals suggest that individuals who suffer from affective disorders benefit from listening to music according to the iso principle. The iso principle comprises listening to music that matches the current mood of patients at first, and then to gradually shift to music that represents a desired mood. Within the current study, we investigate whether the sequence of music with different emotional valence can modulate the emotional state. All participants were healthy adults who underwent a sadness induction via a movie clip. They were subsequently divided into four experimental groups. Each was asked to listen to two pieces of music according to a specific sequence: sad-sad; sad-happy; happy-happy; happy-sad. Participants were prompt to rate their current emotional state at different stages of the experiment: prior to and after the movie clip, as well as after each of the two pieces of music. The frame used for the assessment was the Positive and Negative Affect Schedule and the Self-Assessment Manikin. The results indicate that the movie clip induced sadness. The group of participants who listened to the sad music first and the happy music afterwards ultimately reported a higher positive affect, a higher emotional valence, and a lower negative affect compared with the other groups. However, not all the between-group differences reached significance. We conclude that the sequence of music with different emotional valence affects the current emotional state. The results are generally in line with the iso principle. Directions for future research are presented.

Instruments for the development of the country's export potential

In the context of accelerating globalization and integration trends in the world economic system, countries must make efforts to form their advantageous positions in the international division of labour. This can be realized by optimizing the sphere of foreign economic activity of the state. The purpose of the article is to substantiate the areas of improvement of strategic tools to develop the country's export potential. The authors analyzed the theoretical and methodological basis, namely the essence of the country's export potential, its structural elements of the export potential. The authors analyzed the structure of exports of Kyrgyzstan and Ukraine in 2020, highlighted their common features, shortcomings and explained why the specialization of exports is not optimal. The authors found that a systematic approach to developing a country's development strategy requires a specific sequence in the implementation of this process and proposed an algorithm for implementing the concept of developing the country's export potential, and also proposed calculating a comprehensive indicator of export potential, a more detailed description of which will be presented in further studies.

A Conversation Analytic Study of Laughter in Psychotherapy

<p>This thesis is an investigation into laughter in psychotherapeutic interactions. Conversation analysis was the method used to analyse laughter practices by client and therapist that aid in the business of psychotherapy. Analysing naturally occurring talk is important as it reveals how actions are accomplished, as some past studies on laughter in psychotherapy rely on anecdotal evidence and categorical analysis. Additionally, past psychological literature on laughter can view the phenomenon of laughter as random, and as a by-product of humour. An assumption of conversation analysis is the view of talk being systematic and organised. There is no detail too small that it does not contribute to an interaction (Jefferson, 1985). With this viewpoint in mind conversation analysts have revealed laughter to be an orderly phenomenon that is capable of other actions in talk besides appreciating humour. However, there is a lack of conversation analytical work in laughter during therapy; a gap this thesis sought to address. In particular there were two research questions. If laughter does not have the sole role of appreciating humour, what can it do in psychotherapy? Additionally, past studies in psychotherapy have linked laughter to affiliation in therapy sessions, but do not illustrate the specific sequence of how rapport is achieved in the interaction itself. Psychotherapy can be known as the „talking cure‟ (Perakyla, Antaki, Vehvilainen, & Leudar, 2008), thus, the second question is how does laughter display affiliation in therapeutic talk? Using the fundamental literature of conversation analysis there were two findings regarding laughter in psychotherapy found in this thesis. Firstly, clients would laugh responsively to an action of therapeutic import, the laughter functioned as a marker of dis-preference and an invitation for the therapist to laugh. The therapist would dis-attend the client‟s laughter in order to prompt talk which progressed the therapy from the client. Secondly, therapist could affiliate with the client by display a shared stance towards a matter spoken of by the client. During or after these displays the therapist invited laughter from the client so that the two could laugh together in a further display of shared emotional alignment. These results expanded conversation analytical work on laughter regarding laughter invitations (Jefferson, 1979) and work on psychotherapeutic interactions regarding the prompting of talk (Muntigl, & Hadic Zabala, 2008). The findings also provide empirical evidence for how therapists affiliate with their clients using laughter at the micro-analytical level. The findings of this thesis contribute to psychological, conversation analytical, and psychotherapeutic knowledge on laughter.</p>

A Conversation Analytic Study of Laughter in Psychotherapy

<p>This thesis is an investigation into laughter in psychotherapeutic interactions. Conversation analysis was the method used to analyse laughter practices by client and therapist that aid in the business of psychotherapy. Analysing naturally occurring talk is important as it reveals how actions are accomplished, as some past studies on laughter in psychotherapy rely on anecdotal evidence and categorical analysis. Additionally, past psychological literature on laughter can view the phenomenon of laughter as random, and as a by-product of humour. An assumption of conversation analysis is the view of talk being systematic and organised. There is no detail too small that it does not contribute to an interaction (Jefferson, 1985). With this viewpoint in mind conversation analysts have revealed laughter to be an orderly phenomenon that is capable of other actions in talk besides appreciating humour. However, there is a lack of conversation analytical work in laughter during therapy; a gap this thesis sought to address. In particular there were two research questions. If laughter does not have the sole role of appreciating humour, what can it do in psychotherapy? Additionally, past studies in psychotherapy have linked laughter to affiliation in therapy sessions, but do not illustrate the specific sequence of how rapport is achieved in the interaction itself. Psychotherapy can be known as the „talking cure‟ (Perakyla, Antaki, Vehvilainen, & Leudar, 2008), thus, the second question is how does laughter display affiliation in therapeutic talk? Using the fundamental literature of conversation analysis there were two findings regarding laughter in psychotherapy found in this thesis. Firstly, clients would laugh responsively to an action of therapeutic import, the laughter functioned as a marker of dis-preference and an invitation for the therapist to laugh. The therapist would dis-attend the client‟s laughter in order to prompt talk which progressed the therapy from the client. Secondly, therapist could affiliate with the client by display a shared stance towards a matter spoken of by the client. During or after these displays the therapist invited laughter from the client so that the two could laugh together in a further display of shared emotional alignment. These results expanded conversation analytical work on laughter regarding laughter invitations (Jefferson, 1979) and work on psychotherapeutic interactions regarding the prompting of talk (Muntigl, & Hadic Zabala, 2008). The findings also provide empirical evidence for how therapists affiliate with their clients using laughter at the micro-analytical level. The findings of this thesis contribute to psychological, conversation analytical, and psychotherapeutic knowledge on laughter.</p>