The Growing Problem of Hepatitis C Virus Infection: The Case of West Virginia.

Hepatitis C virus (HCV) is one of the most significant public health problems currently facing the United States, especially in West Virginia. If it is undetected and left untreated, the likelihood of sustaining a treatment response decreases. While early identification has been identified as a critical focus in trying to obtain better health outcomes, new drug treatments appear promising, if somewhat expensive. West Virginia is a predominantly rural state, where the incidence of HCV is 9 times the national average and Medicaid costs for treatment amounted to more than $27 million from 2014 to 2016. The purpose of this study was to conduct a systematic review of the effects of early identification and treatment for patients infected with HCV as it relates to West Virginia. A comprehensive systematic review was limited to 58 articles published from 2008 to 2018 and were in English. Findings from this review identified early detection as the first line of a preventive strategy to help reduce the evolving epidemic and that oral medications could reduce the risk of liver cancer and death. The cost associated with hospitalization of HCV more than tripled from $20 963 in 2005 to $64 867 in 2011 with the average charge per hospitalization at $53 626 due to HVC. The lack of adequate treatment options has led to increasing (and even more expensive) hospital care for untreated HCV. These facts suggest that this state might be facing an expected financial health care crisis due to its increasingly drug-related HCV-infected population.

De novo DNA methylation by DNA methyltransferase 3a controls early effector CD8+ T-cell fate decisions following activation.

DNMT3a is a de novo DNA methyltransferase expressed robustly after T-cell activation that regulates plasticity of CD4(+) T-cell cytokine expression. Here we show that DNMT3a is critical for directing early CD8(+) T-cell effector and memory fate decisions. Whereas effector function of DNMT3a knockout T cells is normal, they develop more memory precursor and fewer terminal effector cells in a T-cell intrinsic manner compared with wild-type animals. Rather than increasing plasticity of differentiated effector CD8(+) T cells, loss of DNMT3a biases differentiation of early effector cells into memory precursor cells. This is attributed in part to ineffective repression of Tcf1 expression in knockout T cells, as DNMT3a localizes to the Tcf7 promoter and catalyzes its de novo methylation in early effector WT CD8(+) T cells. These data identify DNMT3a as a crucial regulator of CD8(+) early effector cell differentiation and effector versus memory fate decisions.

Development and Analysis of a Scale for Meauring Teachers' Sense of Efficacy in Urban Schools (SEUS).

Research in teacher self-efficacy has a long history that can be traced back to Bandura (1986) and has been shown to be linked to teacher performance. This article presents evidence for teacher self-efficacy in urban schools, a construct that is separate from but related to the more general construct of teacher self-efficacy. An instrument was developed and validated by a team of university faculty, urban teachers, and school administrators. The Teachers' Sense of Efficacy in Urban Schools (SEUS) is a 15-item instrument designed to address factors that are important for success in teaching in an urban environment, including working effectively with English language learners, students with disabilities, economically disadvantaged students, cultural diversity, literacy, technology, differentiation, and assessment data. The present study analyzes SEUS on multiple levels, using the Rasch partial credit model.

Clinically relevant orthotopic pancreatic cancer models for adoptive T cell transfer therapy.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is an aggressive tumor. Prognosis is poor and survival is low in patients diagnosed with this disease, with a survival rate of ~12% at 5 years. Immunotherapy, including adoptive T cell transfer therapy, has not impacted the outcomes in patients with PDAC, due in part to the hostile tumor microenvironment (TME) which limits T cell trafficking and persistence. We posit that murine models serve as useful tools to study the fate of T cell therapy. Currently, genetically engineered mouse models (GEMMs) for PDAC are considered a "gold-standard" as they recapitulate many aspects of human disease. However, these models have limitations, including marked tumor variability across individual mice and the cost of colony maintenance. METHODS: Using flow cytometry and immunohistochemistry, we characterized the immunological features and trafficking patterns of adoptively transferred T cells in orthotopic PDAC (C57BL/6) models using two mouse cell lines, KPC-Luc and MT-5, isolated from C57BL/6 KPC-GEMM (KrasLSL-G12D/+p53-/- and KrasLSL-G12D/+p53LSL-R172H/+, respectively). RESULTS: The MT-5 orthotopic model best recapitulates the cellular and stromal features of the TME in the PDAC GEMM. In contrast, far more host immune cells infiltrate the KPC-Luc tumors, which have less stroma, although CD4+ and CD8+ T cells were similarly detected in the MT-5 tumors compared with KPC-GEMM in mice. Interestingly, we found that chimeric antigen receptor (CAR) T cells redirected to recognize mesothelin on these tumors that signal via CD3ζ and 41BB (Meso-41BBζ-CAR T cells) infiltrated the tumors of mice bearing stroma-devoid KPC-Luc orthotopic tumors, but not MT-5 tumors. CONCLUSIONS: Our data establish for the first time a reproducible and realistic clinical system useful for modeling stroma-rich and stroma-devoid PDAC tumors. These models shall serve an indepth study of how to overcome barriers that limit antitumor activity of adoptively transferred T cells.

Mending fences: repairing boundaries through ego state therapy.

Ego state therapy has often been cited as an effective treatment to help repair fragmentation related to posttraumatic stress and dissociative disorders. This article explores how specialized work with ego states can help to clarify and strengthen internal and external boundaries, create greater boundary flexibility, and contribute to containment and self-regulation. Applications of direct and indirect hypnosis to repair boundary issues through ego state therapy are emphasized, and clinical case examples are used to illustrate results.

CES1 Releases Oxylipins from Oxidized Triacylglycerol (oxTAG) and Regulates Macrophage oxTAG/TAG Accumulation and PGE2/IL-1ß Production.

Triacylglycerols (TAGs) are storage forms of fat, primarily found in cytoplasmic lipid droplets in cells. TAGs are broken down to their component free fatty acids by lipolytic enzymes when fuel reserves are required. However, polyunsaturated fatty acid (PUFA)-containing TAGs are susceptible to nonenzymatic oxidation reactions, leading to the formation of oxylipins that are esterified to the glycerol backbone (termed oxTAGs). Human carboxylesterase 1 (CES1) is a member of the serine hydrolase superfamily and defined by its ability to catalyze the hydrolysis of carboxyl ester bonds in both toxicants and lipids. CES1 is a bona fide TAG hydrolase, but it is unclear which specific fatty acids are preferentially released during lipolysis. To better understand the biochemical function of CES1 in immune cells, such as macrophages, its substrate selectivity when it encounters oxidized PUFAs in TAG lipid droplets requires study. We sought to identify those esterified oxidized fatty acids liberated from oxTAGs by CES1 because their release can activate signaling pathways that enforce the development of lipid-driven inflammation. Gaining this knowledge will help fill data gaps that exist between CES1 and the lipid-sensing nuclear receptors, PPARγ and LXRα, which are important drivers of lipid metabolism and inflammation in macrophages. Oxidized forms of triarachidonoylglycerol (oxTAG20:4) or trilinoleoylglycerol (oxTAG18:2), which contain physiologically relevant levels of oxidized PUFAs (<5 mol %), were incubated with recombinant CES1 to release oxylipins and nonoxidized arachidonic acid (AA) or linoleic acid (LA). CES1 hydrolyzed each oxTAG, yielding regioisomers of hydroxyeicosatetraenoic acids (5-, 11-, 12-, and 15-HETE) and hydroxyoctadecadienoic acids (9- and 13-HODE). Furthermore, human THP-1 macrophages with deficient CES1 levels exhibited a differential response to extracellular stimuli (oxTAGs, lipopolysaccharide, and 15-HETE) as compared to those with normal CES1 levels, including enhanced oxTAG/TAG lipid accumulation and altered cytokine and prostaglandin E2 profiles. This study suggests that CES1 can metabolize oxTAG lipids to release oxylipins and PUFAs, and it further specifies the substrate selectivity of CES1 in the metabolism of bioactive lipid mediators. We suggest that the accumulation of oxTAGs/TAGs within lipid droplets that arise due to CES1 deficiency enforces an inflammatory phenotype in macrophages.

Dual IL-6 and CTLA-4 blockade regresses pancreatic tumors in a T cell- and CXCR3-dependent manner.

This study aimed to enhance antitumor immune responses to pancreatic cancer via Ab-based blockade of IL-6 and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). Mice bearing s.c. or orthotopic pancreatic tumors were treated with blocking Abs to IL­6 and/or CTLA-4. In both tumor models, dual IL-6 and CTLA-4 blockade significantly inhibited tumor growth. Additional investigations revealed that dual therapy induced an overwhelming infiltration of T cells into the tumor as well as changes in CD4+ T cell subsets. Dual blockade therapy elicited CD4+ T cells to secrete increased IFN-γ in vitro. Likewise, in vitro stimulation of pancreatic tumor cells with IFN-γ profoundly increased tumor cell production of CXCR3-specific chemokines, even in the presence of IL-6. In vivo blockade of CXCR3 prevented orthotopic tumor regression in the presence of the combination treatment, demonstrating a dependence on the CXCR3 axis for antitumor efficacy. Both CD4+ and CD8+ T cells were required for the antitumor activity of this combination therapy, as their in vivo depletion via Abs impaired outcomes. These data represent the first report to our knowledge of IL-6 and CTLA­4 blockade as a means to regress pancreatic tumors with defined operative mechanisms of efficacy.

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.

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.

A thermodynamic approach to designing structure-free combinatorial DNA word sets.

An algorithm is presented for the generation of sets of non-interacting DNA sequences, employing existing thermodynamic models for the prediction of duplex stabilities and secondary structures. A DNA 'word' structure is employed in which individual DNA 'words' of a given length (e.g. 12mer and 16mer) may be concatenated into longer sequences (e.g. four tandem words and six tandem words). This approach, where multiple word variants are used at each tandem word position, allows very large sets of non-interacting DNA strands to be assembled from combinations of the individual words. Word sets were generated and their figures of merit are compared to sets as described previously in the literature (e.g. 4, 8, 12, 15 and 16mer). The predicted hybridization behavior was experimentally verified on selected members of the sets using standard UV hyperchromism measurements of duplex melting temperatures (T(m)s). Additional experimental validation was obtained by using the sequences in formulating and solving a small example of a DNA computing problem.

Joan of Arc meets Mary Poppins: maternal re-nurturing approaches with male patients in Ego-State Therapy.

Many patients with posttraumatic fragmentation demonstrate a positive response to the corrective possibilities provided through Ego-State Therapy. However, full resolution of presenting symptoms may not occur for individuals with significant childhood histories of parental abuse and neglect without opposite sex, as well as same sex, re-nurturing interventions. This presentation emphasizes the use of maternal re-nurturing methods with men who struggle with the effects of significant attachment deficits in early life. Case examples feature male patients with long-term difficulties in their adult relationships with women that had proved refractory to other therapy methods. Following Ego-State Therapy interventions with maternal symbolic figures, however, these problems improved dramatically. Therapeutic implications for cross-gender re-nurturing with patients who report different types of maternal attachment trauma are explored and discussed.

Comparison of Methods for miRNA Extraction from Plasma and Quantitative Recovery of RNA from Cerebrospinal Fluid.

Interest in extracellular RNA (exRNA) has intensified as evidence accumulates that these molecules may be useful as indicators of a wide variety of biological conditions. To establish specific exRNA molecules as clinically relevant biomarkers, reproducible recovery from biological samples and reliable measurements of the isolated RNA are paramount. Toward these ends, careful and rigorous comparisons of technical procedures are needed at all steps from sample handling to RNA isolation to RNA measurement protocols. In the investigations described in this methods paper, RT-qPCR was used to examine the apparent recovery of specific endogenous miRNAs and a spiked-in synthetic RNA from blood plasma samples. RNA was isolated using several widely used RNA isolation kits, with or without the addition of glycogen as a carrier. Kits examined included total RNA isolation systems that have been commercially available for several years and commonly adapted for extraction of biofluid RNA, as well as more recently introduced biofluids-specific RNA methods. Our conclusions include the following: some RNA isolation methods appear to be superior to others for the recovery of RNA from biological fluids; addition of a carrier molecule seems to be beneficial for some but not all isolation methods; and quantitative recovery of RNA is observed from increasing volumes of cerebrospinal fluid.

Women's sexual satisfaction as a predictor of well-being in same-sex versus mixed-sex relationships.

Structural equation modelling was used to assess the strengths of the links between sexual satisfaction and self-reported (a) relationship well-being, (b) mental health, and (c) physical health for women in same-sex (i.e., homosexual, n = 114) versus mixed-sex (i.e., heterosexual, n = 208) relationships. Participants came from a large-scale Internet study. Sexual satisfaction was found to be an extremely strong predictor of relational well-being, a strong predictor of mental health, and a weak to moderately strong predictor of physical health. A two-group comparison model indicated that the strength of these links was the same, regardless of whether the women were in a sexual relationship with a man or with another woman.