Gazing Back, Playing Forward: Contemporary Psychoanalytic Musings on the Relational Essence of Hypnotherapeutic Action.

To build bridges between hypnosis and contemporary psychoanalysis, this article addresses how hypnosis, when used in psychotherapy, facilitates curative action through its relational essence. The author's extensive experience with hypnosis, psychotherapy, and psychoanalysis orient the narrative toward the unconscious patient-therapist interaction, with particular attention paid to the ethics of the inherent hypnotic seduction. Whether used primarily in relief-oriented ways or geared toward more transformative therapeutic aims, powerful unconscious factors are in play for both patient and therapist and are explicated to illustrate the interactive and frequently unformulated, intersubjective factors that facilitate effective, psychotherapeutic hypnosis. Consequently, therapists attuned to such intersubjective dynamics can make use of their own internal mental activities to understand a patient's current state of mind and level of developmental functioning, and thereby subsequently formulate mutative interventions. For instance, because hypnotizability reflects the ability to play in imaginative space, the regression promoted in hypnotherapy may activate both an illusion of omnipotence and its optimal disillusionment through the relational context. This requires going beyond more traditional, procedural ways of bifurcating hypnotic interventions as being either direct or indirect and instead further distinguish hypnotic interventions in accordance with their maternal and paternal relational dimensions. Arguably, then, the skillful hypnotherapist needs to maintain a coupling interplay between the maternal, maximally receptive and the paternal, more active modes of functioning within hypnotic play space.

The Generative Presence of Relatedness.

In this guest editorial, the authors introduce a special issue of the American Journal of Clinical Hypnosis that focuses on relational factors of hypnosis in psychotherapy. The authors have invited a number of esteemed colleagues to comment on aspects of the therapeutic relationship, and how it informs and influences the processes, techniques, and outcomes of hypnosis and therapy. In addition to summarizing each of these articles, this article analyzes the major relational themes that present across the articles.

In the Intersubjective Space: Hypnosis Through a Neuropsychological Lens.

This article explores five interwoven principles about relationship that impact on attentional focus as it relates to the practice of clinical hypnosis. It first reviews how relationship is an irreducible feature of life that greatly predates the arrival of human beings. Second, it describes brain structures that, from an evolutionary perspective, appeared relatively recently, and the neuropsychological abilities those structures confer on human relationships. Third, it links those social brain structures to trance, an inborn response to novelty that is an important feature of our adaptive learning capacity. It further suggests that narrative is a multilevel concept that is deeply embodied and constitutes the sorbate from which hypnotic interactions can draw their rich impact. Finally, the article posits that hypnosis represents a skill set through which attuned clinicians engage in co-creative dances with clients, where the choreography of their interaction attends and responds to spontaneously arising and/or deliberately seeded opportunities for adaptive change. Reconstructed descriptions of interactions with clients are provided to illustrate the application of the principles presented.

Hypnosis and The Therapeutic Relationship: Relational Factors of Hypnosis in Psychotherapy.

Over the years, the field of hypnosis has often given more attention to the state and procedural factors of hypnosis than the relational ones. In an attempt to address this imbalance, the 60th annual meeting of the American Society of Clinical Hypnosis (ASCH) had as its theme "Hypnosis and the Treatment Relationship." A centerpiece of this meeting was a collegial discussion among a panel of psychologists with expertise in relational hypnotherapy. The panel addressed several pertinent questions: (1) What are the healing qualities of relationship in psychotherapy? (2) What makes hypnosis relational in nature? (3) How do relational factors of hypnosis resemble healthy attachment processes and recapitulate stalled developmental maturation? (4) How does relationally informed hypnotherapy influence and strengthen the relationship, process, and outcome of psychotherapy? This article summarizes the factors that led to the creation of this panel; provides an edited transcript of this panel discussion, along with additional commentary on several key points raised; and concludes with a summary of the main themes and recommendations for further clinical practice and study.

Dancing in the In-Between: Hypnosis, Transitional Space, and Therapeutic Action.

This article develops the idea that hypnosis is an interactive phenomenon occurring in a relational matrix. A tripartite model for explicating this relational matrix is presented, which includes a discussion of transference, contemporary relationship factors, and the interaction of these to produce a sense of therapeutic alliance. These relationship factors are central to the therapeutic action of hypnosis as a vehicle to potentiate change and growth. The unique and specific contribution of each of these factors to the process of hypnotherapy and to therapeutic action is examined. Phenomenologically, this relational interaction is conceptualized as occurring in transitional space, shaped by processes of regression and attunement. From this perspective, the hypnotherapist is viewed as a kind of transitional object whose empathic presence contains and facilitates those interactive phenomena which evoke and balance the transferential and contemporary aspects of the relationship and which allow for uniquely evocative developmentally focused interventions in trance work. Several examples are presented from an ongoing case that demonstrate how these relational variables shape the hypnotherapeutic process and how they can be used for uncovering and self-examination, for structural maturation, for affect regulation, and for emerging ego mastery. The therapeutic action demonstrated relies on hypnotic interventions rooted in the various components of the relational matrix made possible by the clinician's awareness of and attunement to these and by hisor her informed and sensitive management of them and of his or her own intersubjectivity. Specific strategies are presented via these examples to effectively utilize this experience in the service of treatment goals.

It Takes More Than Two to Tango: Building Secure Attachment Through Hypnotic and Ego-State Relationships.

This article explores how hypnotic strategies can be used within a polyvagal science framework to help create more secure attachment within the therapeutic relationship, as well as within the client in terms of ego-state relationships. Principles of safety and connection are emphasized, along with specific strategies to access the attachment circuits of the ventral vagal system, including the necessity of being present with the client without agenda. Uses of hypnosis related to safety and connection and methods to work with the center core self to facilitate empowerment, self-cohesion, and conflict-free experience are also reviewed. From an ego-state therapy perspective, a discussion of hypnosomatic approaches to connect with preverbal, nonverbal, and somatic aspects of self to accomplish developmental repair and facilitate secure attachment is also offered, along with case examples. A three-step model, which attempts to integrate polyvagal, somatic, and hypnotic approaches, is offered by the author to help structure corrective experiences for clients with trauma.

Relationship Factors in the Theater of the Imagination: Hypnosis With Children and Adolescents.

This article examines the spatial and social nature of human relationships with children and adolescents in clinical hypnosis. Beginning with the unique way in which the phenomenon of rapport is intrinsic to the therapeutic uses of hypnosis and is distinct among other therapies, the stage is set for the importance of relational hypnosis. Through the use of case vignettes that illustrate developmental imperatives, relationship factors influencing the clinical interaction are demonstrated in practice. These include transference and countertransference, safety, embodiment, novelty, creativity, respect, trust, equality, being with, loving responses, synchronicity, and empathy. Hypnotic relating exists in a framework through which absorption in play and imagination evokes the child's resources and suggestions are made. In this receptive stance the personalized suggestions lead to an environment for positive change.

Longing and Fear: The Ambivalence About Having a Relationship in Psychotherapy.

It is unquestioned that reaching the hypnotic state is helped along by relational factors and that, conversely, relational experiences can be deepened through hypnosis. It is also true that deepening the experience of being in a relationship with another person is neither comfortable nor indicated for every patient or therapist. Most humans feel ambivalent about closeness. People vary in their desire for and their skill in sustaining mature intimacy. When we move along the continuum from rudimentary notions about relational factors in psychotherapy, such as rapport, to complex concepts, such as enactments, we move along a corresponding continuum of increasing need for specialized training, supervised experience, and personal therapy. The field of psychotherapy has been plagued from its inception by not knowing what to do with the tensions that emerge when two people listen to and look at each other. Avoiding relational factors may be a very human response to a very daunting matter.

Trends and Health Risks of Dissolved Heavy Metal Pollution in Global River and Lake Water from 1970 to 2017.

Heavy metal pollution in surface water is a global environmental problem. This study analyzed the trends, health risks, and sources of eight dissolved heavy metal species in river and lake water across five continents (Africa, Asia, Europe, North America, and South America; Oceania was excluded owing to a lack of data) for the period 1970-2017. We wanted to assess the effects of various implemented countermeasures to pollution and to determine those that could be adopted worldwide. Collectively, the water system showed increasing trends for Cd, Cr, Cu, Ni, Mn, and Fe and decreasing trends for Pb and Zn. The mean dissolved concentrations of most heavy metals were highest in Asia and lowest in Europe. Most heavy metals had low non-carcinogenic risks over this period. The cancer risks associated with Pb were lower than the hazardous level on all five continents over the five decades, whereas the cancer risks related to Cr exceeded the hazardous level in the 1970s, 2000s, and 2010s, and in Africa, Asia, and North America over the entire period. Mining and manufacturing were consistently found to be critical sources of metal pollution from 1970 to 2017. However, the heavy metal sources differed significantly by continent, with waste discharge and rock weathering dominant in Africa; mining and manufacturing, along with rock weathering, are dominant in Asia and South America; fertilizer and pesticide use, along with rock weathering, are dominant in North America; and mining and manufacturing, waste discharge, and rock weathering are dominant in Europe. Global trends in the metal loadings in water and in relevant pollution-control measures suggest that countermeasures in Europe have successfully controlled heavy metal pollution. The successful measures include implementing rigorous standards for metal emissions, limiting the metal concentrations in products, and rigorously treating metal-contaminated waste. Therefore, the measures implemented in Europe should be extended worldwide to treat heavy metal pollution in water.

Anaerobic Microbial Degradation of Polycyclic Aromatic Hydrocarbons: A Comprehensive Review.

Polycyclic aromatic hydrocarbons (PAHs) are a class of hazardous organic contaminants that are widely distributed in nature, and many of them are potentially toxic to humans and other living organisms. Biodegradation is the major route of detoxification and removal of PAHs from the environment. Aerobic biodegradation of PAHs has been the subject of extensive research; however, reports on anaerobic biodegradation of PAHs are so far limited. Microbial degradation of PAHs under anaerobic conditions is difficult because of the slow growth rate of anaerobes and low energy yield in the metabolic processes. Despite the limitations, some anaerobic bacteria degrade PAHs under nitrate-reducing, sulfate-reducing, iron-reducing, and methanogenic conditions. Anaerobic biodegradation, though relatively slow, is a significant process of natural attenuation of PAHs from the impacted anoxic environments such as sediments, subsurface soils, and aquifers. This review is intended to provide comprehensive details on microbial degradation of PAHs under various reducing conditions, to describe the degradation mechanisms, and to identify the areas that should receive due attention in further investigations.

ADAMTS Proteins: Concepts, Challenges, and Prospects.

The ADAMTS superfamily comprises secreted metalloproteases (ADAMTS proteases) as well as structurally related secreted glycoproteins that lack catalytic activity (ADAMTS-like proteins). Members of both families participate in diverse morphogenetic processes during embryonic development, and connective tissue maintenance and hemostasis in the adult. Several ADAMTS proteins are heavily implicated in genetic and acquired human and animal disorders. Despite these indicators of a profound biological and medical importance, detailed knowledge about their molecular structures, substrates, biological pathways, and biochemical mechanisms is significantly limited by unique intrinsic characteristics, which have led to several technical challenges. As a group, they are larger, more heavily modified, and harder to purify than other secreted proteases. In addition, idiosyncratic aspects of individual members are deserving of further investigation but can complicate their analysis. Here, some of the key concepts, challenges, and prospects in ADAMTS research are discussed in the context of the knowledge accumulated over the past two decades. Individual chapters in this volume of Methods in Molecular Biology provide practical solutions for surmounting these challenges. Since the biology of a protease is actually the biology of its substrates, there is considerable emphasis on purification of recombinant ADAMTS proteins, identification of their substrates and assays for their proteolytic activity.

Inhibition of ADAMTS1 Expression by Lentiviral CRISPR/Cas9 Gene Editing Technology.

The continuous improvement of gene editing tools has allowed a major revolution in biological sciences. Although a variety of gain and loss-of-function approaches have been widely used for the last decades, some limitations arose from non-specific targeting or lack of complete inhibition of the gene of interest. CRISPR/Cas9 editing technology introduced new and significant advantages because it can directly modify the gene of interest and completely blocks its expression.In the context of cancer studies, the heterogeneity of the tumor microenvironment requires comprehensive approaches to unveil the contribution of multiple genes. For example, a deeper understanding of the biology of proteases such as ADAMTS (a disintegrin and metalloproteinase with thrombospondin type 1 motifs) will improve our perspective of complex phenomena affected by extracellular matrix remodeling, including embryonic development, angiogenesis, immune infiltration, metastasis, and tumor plasticity. Here, we present a method using CRISPR/Cas9 technology to inhibit the expression of the representative ADAMTS1 in cancer cells. Following the first steps of gene edition, we pursue further selection of silenced cells and provide a detailed description of sequence analysis and validation assays. This method leads to inactivation of ADAMTS1 in cancer cells, providing a relevant biological tool that will allow subsequent in vivo and in vitro ADAMTS1 functional analysis.

Analyzing the Effects of O-Fucosylation on Secretion of ADAMTS Proteins Using Cell-Based Assays.

Metalloproteinases of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin type 1 repeats) superfamily are extensively modified with glycan moieties. Glycosylation occurs as these enzymes are trafficked through the endoplasmic reticulum (ER) and Golgi apparatus on their way to the extracellular space and includes N-linked glycosylation, O-linked fucosylation and C-linked mannosylation. This chapter focuses on O-linked fucose, which is added to properly folded thrombospondin type 1 repeats (TSRs) in the ER by protein O-fucosyltransferase 2 (POFUT2) and elongated to a Glucoseß1-3Fucose disaccharide by ß3-glucosyltransferase (B3GLCT). Knockout of POFUT2 results in embryonic lethality in mice, and inactivating mutations in B3GLCT cause Peters plus syndrome, a congenital disorder of glycosylation in humans. Addition of the disaccharide by POFUT2 and B3GLCT stabilizes folded TSRs, enhancing folding in the ER and secretion efficiency of several ADAMTS proteins from cells. Thus, POFUT2 and B3GLCT both function as an ER quality control pathway for folding of TSRs in ADAMTS proteins. In this chapter we describe in detail the methods developed to analyze secretion defects of ADAMTS proteins upon loss of either POFUT2 or B3GLCT. The methods described include creation of CRISPR/Cas9-mediated knockout cell lines of POFUT2 and B3GLCT and use of these cell lines to analyze and quantify secretion defects of ADAMTS proteins.

Challenges and Solutions for Purification of ADAMTS Proteases: An Overview.

ADAMTS are secreted metalloproteinases implicated in many key biological processes. The 19 different members of this family share an identical domain composition at the level of their amino-terminal portion, whereas the identity and number of the domains forming their carboxy-terminal half are divergent and define distinct ADAMTS subfamilies. Due to their large size, extensive glycosylation, the presence of specific domains, their tendency to form aggregates, their relatively low abundance in tissues and the presence of many disulfide bonds, ADAMTS are very hard to isolate, express, and purify, as either native or recombinant active enzymes. This chapter provides an overview of critical steps to take into account when obtaining these proteases for biochemical and functional investigation.

Purification of Native or Recombinant ADAMTS2, and Procollagen I Cleavage Assay.

ADAMTS constitute a family of 19 secreted metalloproteinases involved in diverse physiopathological conditions. Most of their roles first emerged from analysis of spontaneous human and animal mutations or genetically engineered animals. However, the involved mechanisms and the full repertoire of their functions are still largely unrecognized, in part because they are difficult to produce and purify as recombinant active enzymes. Here we describe protocols, tips, and tricks specifically regarding ADAMTS2, 3, and 14 but still relevant for other ADAMTS.

ADAMTS7: Recombinant Protein Expression and Purification.

ADAMTS7 is a secreted protease that is predominantly expressed in tissues of the cardiovascular system and tendon. Although recent evidence suggests that it may play a role in the etiology of coronary artery disease, its physiological function and substrates are unknown. The enzyme undergoes extensive posttranslational modifications, including chondroitin sulfate attachment, N and O-linked glycosylation, and a two-step activation process. For the benefit of scientists who study the function of ADAMTS7 and its role in disease, this chapter provides an introduction to the chemical and functional properties of the various ADAMTS7 domains, as well as a protocol for the recombinant expression and purification of ADAMTS7.

Purification and Activity Determination of ADAMTS-4 and ADAMTS-5 and Their Domain Deleted Mutants.

A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs-4 (ADAMTS-4) and ADAMTS-5 are zinc-dependent metalloproteinases that are involved in the maintenance of cartilage extracellular matrix (ECM) and are currently considered the major aggrecanases in the development of osteoarthritis. In this chapter we describe the establishment and cultivation of cell lines expressing ADAMTS-4,-5 and their domain deletion mutants; the collection of medium containing expressed ADAMTS-4,-5; the subsequent purification of this medium through anti-FLAG affinity chromatography; and the characterization of ADAMTS-4,-5 activity using synthetic Förster resonance energy transfer (FRET) peptide substrates.