Phosphorylation of Ser1928 mediates the enhanced activity of the L-type Ca2+ channel Cav1.2 by the ß2-adrenergic receptor in neurons.

The L-type Ca2+ channel Cav1.2 controls multiple functions throughout the body including heart rate and neuronal excitability. It is a key mediator of fight-or-flight stress responses triggered by a signaling pathway involving ß-adrenergic receptors (ßARs), cyclic adenosine monophosphate (cAMP), and protein kinase A (PKA). PKA readily phosphorylates Ser1928 in Cav1.2 in vitro and in vivo, including in rodents and humans. However, S1928A knock-in (KI) mice have normal PKA-mediated L-type channel regulation in the heart, indicating that Ser1928 is not required for regulation of cardiac Cav1.2 by PKA in this tissue. We report that augmentation of L-type currents by PKA in neurons was absent in S1928A KI mice. Furthermore, S1928A KI mice failed to induce long-term potentiation in response to prolonged theta-tetanus (PTT-LTP), a form of synaptic plasticity that requires Cav1.2 and enhancement of its activity by the ß2-adrenergic receptor (ß2AR)-cAMP-PKA cascade. Thus, there is an unexpected dichotomy in the control of Cav1.2 by PKA in cardiomyocytes and hippocampal neurons.

Phosphorylation of Cav1.2 on S1928 uncouples the L-type Ca2+ channel from the ß2 adrenergic receptor.

Agonist-triggered downregulation of ß-adrenergic receptors (ARs) constitutes vital negative feedback to prevent cellular overexcitation. Here, we report a novel downregulation of ß2AR signaling highly specific for Cav1.2. We find that ß2-AR binding to Cav1.2 residues 1923-1942 is required for ß-adrenergic regulation of Cav1.2. Despite the prominence of PKA-mediated phosphorylation of Cav1.2 S1928 within the newly identified ß2AR binding site, its physiological function has so far escaped identification. We show that phosphorylation of S1928 displaces the ß2AR from Cav1.2 upon ß-adrenergic stimulation rendering Cav1.2 refractory for several minutes from further ß-adrenergic stimulation. This effect is lost in S1928A knock-in mice. Although AMPARs are clustered at postsynaptic sites like Cav1.2, ß2AR association with and regulation of AMPARs do not show such dissociation. Accordingly, displacement of the ß2AR from Cav1.2 is a uniquely specific desensitization mechanism of Cav1.2 regulation by highly localized ß2AR/cAMP/PKA/S1928 signaling. The physiological implications of this mechanism are underscored by our finding that LTP induced by prolonged theta tetanus (PTT-LTP) depends on Cav1.2 and its regulation by channel-associated ß2AR.

Changing Beliefs about Trauma: A Qualitative Study of Cognitive Processing Therapy.

BACKGROUND: Controlled qualitative methods complement quantitative treatment outcome research and enable a more thorough understanding of the effects of therapy and the suspected mechanisms of action. AIMS: Thematic analyses were used to examine outcomes of cognitive processing therapy (CPT) for posttraumatic stress disorder (PTSD) in a randomized controlled trial of individuals diagnosed with military-related PTSD (n = 15). METHOD: After sessions 1 and 11, participants wrote "impact statements" describing their appraisals of their trauma and beliefs potentially impacted by traumatic events. Trained raters coded each of these statements using a thematic coding scheme. RESULTS: An analysis of thematic coding revealed positive changes over the course of therapy in participants' perspective on their trauma and their future, supporting the purported mechanisms of CPT. CONCLUSION: Implications of this research for theory and clinical practice are discussed.

Pleiotropic effects of deubiquitinating enzyme Ubp5 on growth and pathogenesis of Cryptococcus neoformans.

Ubiquitination is a reversible protein modification that influences various cellular processes in eukaryotic cells. Deubiquitinating enzymes remove ubiquitin, maintain ubiquitin homeostasis and regulate protein degradation via the ubiquitination pathway. Cryptococcus neoformans is an important basidiomycete pathogen that causes life-threatening meningoencephalitis primarily in the immunocompromised population. In order to understand the possible influence deubiquitinases have on growth and virulence of the model pathogenic yeast Cryptococcus neoformans, we generated deletion mutants of seven putative deubiquitinase genes. Compared to other deubiquitinating enzyme mutants, a ubp5Δ mutant exhibited severely attenuated virulence and many distinct phenotypes, including decreased capsule formation, hypomelanization, defective sporulation, and elevated sensitivity to several external stressors (such as high temperature, oxidative and nitrosative stresses, high salts, and antifungal agents). Ubp5 is likely the major deubiquitinating enzyme for stress responses in C. neoformans, which further delineates the evolutionary divergence of Cryptococcus from the model yeast S. cerevisiae, and provides an important paradigm for understanding the potential role of deubiquitination in virulence by other pathogenic fungi. Other putative deubiquitinase mutants (doa4Δ and ubp13Δ) share some phenotypes with the ubp5Δ mutant, illustrating functional overlap among deubiquitinating enzymes in C. neoformans. Therefore, deubiquitinating enzymes (especially Ubp5) are essential for the virulence composite of C. neoformans and provide an additional yeast survival and propagation advantage in the host.

Cryptococcus neoformans requires a functional glycolytic pathway for disease but not persistence in the host.

Cryptococcus neoformans is an important fungal pathogen of immunocompromised individuals, with a close relative, Cryptococcus gattii, emerging as a serious threat for the immunocompetent. During initial infection, C. neoformans colonizes the airspaces of the lungs, resulting in pneumonia, and subsequently migrates to the central nervous system (CNS). We sought to understand fungal carbon utilization during colonization of these fundamentally different niches within the host, in particular the roles of gluconeogenesis and glycolysis. We created mutants at key points in the gluconeogenesis/glycolysis metabolic pathways that are restricted for growth on lactate and glucose, respectively. A phosphoenolpyruvate carboxykinase mutant (the pck1Δ mutant), blocked for entry of 2- and 3-carbon substrates into gluconeogenesis and attenuated for virulence in a murine inhalation model, showed wild-type (WT) persistence in a rabbit cerebrospinal fluid (CSF) model of cryptococcosis. Conversely, both the pyruvate kinase (pyk1Δ) and the hexose kinase I and II (hxk1Δ/hxk2Δ) mutants, which show impaired glucose utilization, exhibited severely attenuated virulence in the murine inhalation model of cryptococcosis and decreased persistence in the CNS in both the rabbit CSF and the murine inhalation models while displaying adequate persistence in the lungs of mice. These data suggest that glucose utilization is critical for virulence of C. neoformans and persistence of the yeast in the CNS.

A feasibility study comparing two chiropractic protocols in the treatment of patellofemoral pain syndrome.

OBJECTIVE: The purpose of this pilot study was to determine the feasibility of (1) using the existing clinic, clinicians, interns, faculty, and staff from our college in conducting all components of a planned randomized controlled clinical trial; (2) successfully recruiting patients with patellofemoral pain syndrome (PFPS); and (3) consistently, effectively, and safely implementing the study protocols and therapy. METHODS: Diagnostic, treatment, and blind assessment procedures were carried out while recruitment and administrative techniques for managing long-term storage of data and files were developed. Thirty-one patients were randomized into a local manipulative group (group A) or to a full kinetic chain manipulative therapy group (group B), each combined with exercise and soft tissue treatment. The Anterior Knee Pain Scale, visual analog scale, and Patient Satisfaction Scale were used. RESULTS: All phases of the feasibility study including use of the clinic, staff, recruitment techniques, treatment protocols, data collection, input, and analysis were effectively and safely carried out. CONCLUSIONS: A feasibility study investigating the ability to conduct a randomized controlled trial of a manipulative therapy protocol for PFPS using available chiropractic college infrastructure was accomplished. A fully powered PFPS trial is feasible and merited.

Change in posttraumatic stress disorder symptoms: do clinicians and patients agree?

This study assessed the longitudinal association between clinician and patient ratings of posttraumatic stress disorder (PTSD) symptoms over the course of 2 different randomized clinical trials of veterans with chronic PTSD. One trial, the Department of Veterans Affairs Cooperative Study 420 (CSP 420; N = 360) compared trauma-focused and present-centered group therapies, and the 2nd trial compared cognitive processing theory and a waitlist control condition (N = 60). Linear mixed effects modeling revealed significant associations between clinician ratings (Clinician-Administered PTSD Scale; CAPS; D. D. Blake et al., 1990) and patient ratings (Posttraumatic Stress Disorder Checklist; PCL; F. W. Weathers, B. T. Litz, J. A. Herman, J. A. Huska, & T. M. Keane, 1993) in total and symptom clusters of PTSD. Contrary to hypothesis, the amount of change on the CAPS ranged from .75 to .82 standard deviations for every 1 standard deviation change on the PCL. The CAPS and PCL were more closely associated in the trauma-focused vs. present-centered treatment condition in CSP 420, and especially regarding hyperarousal symptoms. When comparing categorization of clinically significant change on the CAPS and PCL, the authors found no differences in the percentages of agreement between clinicians and patients in improvement and exacerbation. The value of multimodal assessment of PTSD treatment outcomes is discussed.

Tandem optimization of target activity and elimination of mutagenic potential in a potent series of N-aryl bicyclic hydantoin-based selective androgen receptor modulators.

Pharmacokinetic studies in cynomolgus monkeys with a novel prototype selective androgen receptor modulator revealed trace amounts of an aniline fragment released through hydrolytic metabolism. This aniline fragment was determined to be mutagenic in an Ames assay. Subsequent concurrent optimization for target activity and avoidance of mutagenicity led to the identification of a pharmacologically superior clinical candidate without mutagenic potential.

The role of emotional functioning in military-related PTSD and its treatment.

This two-part study investigated the cross-sectional and across-treatment relationships among measures of emotional functioning and posttraumatic stress disorder (PTSD) symptomatology for veterans receiving specialized treatment for military-related PTSD. Cross-sectional analyses revealed overlap among emotion regulation, affective control, depressive symptoms, and PTSD symptoms at pre-treatment. In regression analyses, fear of losing affective control was most predictive of PTSD symptoms. Bivariate analyses of residualized change scores showed that changes in emotion regulation and fear of losing affective control were associated with changes in PTSD and depressive symptoms across treatment. Regression analyses revealed that changes in fear of losing affective control most strongly predicted changes in PTSD and depressive symptoms. Theoretical and clinical implications are discussed, including an understanding of the differences between emotion regulation and affective control processes. Future research directions are offered, including improved measurement of emotional functioning and longitudinal research delineating the likely bi-directional relationship between emotional functioning and PTSD.

A review of interpersonal-psychodynamic group psychotherapy outcomes for adult survivors of childhood sexual abuse.

This article discusses the use of interpersonal-psychodynamic group treatments for survivors of childhood sexual abuse. Empirically based interpersonal-psychodynamic group outcome studies are reviewed according to efficacy and effectiveness models of treatment outcome research. To facilitate the evaluation and comparison of clinically significant change across studies, effect size indicators were calculated when descriptive data were reported. Findings suggest that interpersonal-psychodynamic group therapy is an effective treatment approach for many survivors of childhood sexual abuse. Implications for future research are considered.

Emotional deficits in military-related PTSD: an investigation of content and process disturbances.

To expound on the nature of emotional deficits in PTSD, the current study investigated the relationships among emotion content and process variables and PTSD symptomatology in a sample of 85 veterans with military-related trauma. Alexithymic externally oriented thinking and negative affectivity emerged as the most consistent predictors of PTSD symptoms; however, depression was the only variable associated with emotional numbing. Theoretical and clinical implications of these findings are discussed, as well as future research directions including the collateral and clinician assessment of emotional functioning, use of other process measures, and inclusion of various control groups.

Psychological assessment of adult survivors of childhood sexual abuse within a naturalistic clinical sample.

This study investigates the long-term effects of childhood sexual abuse (CSA). Differences between abused and nonabused individuals in psychiatric symptomatology, interpersonal functioning, social and occupational functioning, personality dynamics, and therapeutic alliance were examined. The relationship between abuse severity and long-term effects was also analyzed. Data were gathered from 51 patients seeking individual psychotherapy at a community outpatient clinic. Findings suggested that CSA survivors tend to experience greater psychiatric distress and poorer interpersonal functioning than nonabused clinical controls. No significant differences were found in social and occupational functioning or in alliance developed by the end of the therapeutic assessment process. Abuse severity was significantly related to increased symptomatology and poorer interpersonal functioning. Findings support and extend existing literature and are especially useful for clinicians working with abuse survivors.