Fluorescence-based incision assay for human XPF-ERCC1 activity identifies important elements of DNA junction recognition

The structure-specific endonuclease activity of the human XPF–ERCC1 complex is essential for a number of DNA processing mechanisms that help to maintain genomic integrity. XPF–ERCC1 cleaves DNA structures such as stem–loops, bubbles or flaps in one strand of a duplex where there is at least one downstream single strand. Here, we define the minimal substrate requirements for cleavage of stem–loop substrates allowing us to develop a real-time fluorescence-based assay to measure endonuclease activity. Using this assay, we show that changes in the sequence of the duplex upstream of the incision site results in up to 100-fold variation in cleavage rate of a stem-loop substrate by XPF-ERCC1. XPF–ERCC1 has a preference for cleaving the phosphodiester bond positioned on the 3′-side of a T or a U, which is flanked by an upstream T or U suggesting that a T/U pocket may exist within the catalytic domain. In addition to an endonuclease domain and tandem helix–hairpin–helix domains, XPF has a divergent and inactive DEAH helicase-like domain (HLD). We show that deletion of HLD eliminates endonuclease activity and demonstrate that purified recombinant XPF–HLD shows a preference for binding stem–loop structures over single strand or duplex alone, suggesting a role for the HLD in initial structure recognition. Together our data describe features of XPF–ERCC1 and an accepted model substrate that are important for recognition and efficient incision activity

Advocating for Holistic Admissions Review: Implications for Master’s Level Counselor Education Programs

Positioning diversity at the forefront of counselor education has been an ongoing movement for well over the last decade. Attention has been given to considering recruitment and retention practices, with CACREP requiring counselor education programs to show evidence of ways this is enacted. Building on this work, counselor educators have brought needed attention to questioning how structural inequity permeates policies and procedures within counselor education programs. A critical consideration of counselor education admissions practices through an equity lens has been lacking in the literature. In this article, the authors make the case for the use of holistic admissions review as an equitable practice within counselor education

Clinical Mediation: Advocating for a New Role

Mediation training standards are inconsistent and fail to focus on the pivotal skill set of addressing, understanding, and working with underlying emotions that shape conflict and may prevent mutual goal attainment. The purpose of this article is to explore skills essential to counseling and mediation and advocate for the potential benefits of formally training counselors to facilitate the process of mediation and conflict resolution

Clinical Mediation: Advocating for a New Role

Mediation training standards are inconsistent and fail to focus on the pivotal skill set of addressing, understanding, and working with underlying emotions that shape conflict and may prevent mutual goal attainment. The purpose of this article is to explore skills essential to counseling and mediation and advocate for the potential benefits of formally training counselors to facilitate the process of mediation and conflict resolution

Duloxetine for the management of fibromyalgia syndrome

Fibromyalgia syndrome (FMS) is a widespread pain condition associated with a wide range of additional symptoms including fatigue, insomnia, depression, anxiety and stiffness. Duloxetine is one of three medications currently FDA approved for use in FMS management. Duloxetine is a mixed serotonin and norepinephrine reuptake inhibitor (SNRI) that functions by increasing central nervous system levels of serotonin and norepinephrine. This review is a primer on use of duloxetine in FMS management and includes information on pharmacology and pharmacokinetics, a review of the three duloxetine FMS treatment trials currently in publication, a discussion of the safety and tolerability of duloxetine, and patient-focused perspectives on duloxetine use in FMS management. Duloxetine has proven efficacy in managing pain and mood symptoms in adult FMS patients with and without major depressive disorder. However, due to side effects, duloxetine must be used with caution in patients with fatigue, insomnia, gastrointestinal complaints, headache, cardiovascular disease, bleeding-risk, and in those 24 years of age and younger due to risk of suicidality. Duloxetine use should be avoided in patients with liver disease or alcoholics. As with all medications, duloxetine is best used as part of an individualized regimen that includes nonpharmacologic modalities of exercise, education and behavioral therapies

Fostering Leadership Capacity in Three South Carolina High Schools: An Exploratory Study

Some states include fostering teacher leadership in principal evaluations, so principals need to provide opportunities and mentor or coach their staffs to assume leadership roles. Yet, less is known about the specific ways principals develop the leadership capacities of others. In this exploratory study we examined how principals in three South Carolina high schools intentionally fostered the capacities of leaders to enhance their schools’ organizational capacities. Our research questions were: 1) How are emerging leaders identified and selected by their principals? 2) How do principals foster leadership capacity in these leaders? Data collected for this multi-site qualitative study consisted of artifacts and semi-structured 60-90 minute interviews. All interviews were transcribed and analyzed by the research team inductively and deductively using NVivo 10. Our findings indicated that principals selected leaders who: indicated a desire to go into formal leadership roles; had good interpersonal skills and rapport with students and parents; demonstrated persistence and willingness; were effective teachers; had strong personalities; and had a particular skill or knowledge. Our findings for the second question indicate that principals fostered leadership by: mentoring and coaching others to leadership; scaffolding opportunities to lead; and trusting leaders to make the right decision

CAML Does Not Modulate Tetherin-Mediated Restriction of HIV-1 Particle Release

Background: Tetherin/BST-2 is a recently-identified potent restriction factor in human cells that restricts HIV particle release following particle formation and budding at the plasma membrane. Vpu counteracts tetherin’s restriction of particle release in a manner that has not yet been fully defined. We recently identified calcium-modulating cyclophilin ligand (CAML) as a Vpu-interacting protein that also restricts particle release. We hypothesized that CAML may act to enhance tetherinmediated restriction of particle release and thereby explain how two distinct factors could be responsible for Vpuresponsive restriction. Methodology/Principal Findings: Endogenous levels of tetherin in human cells correlated well with their restriction pattern and responsiveness to Vpu, while levels of cellular CAML protein did not. Tetherin but not CAML was inducible by interferon in a wide variety of human cells. Stable depletion of human CAML in restrictive HeLa cells had no effect on cell surface levels of tetherin, and failed to relieve tetherin-mediated restriction. Stable depletion of tetherin from HeLa cells, in contrast, rendered HeLa cells permissive and Vpu-unresponsive. Tetherin but not CAML expression in permissive human cells rendered them restrictive and Vpu responsive. Depletion of CAML had no influence on cell surface levels of tetherin. Conclusions/Significance: We conclude that tetherin restricts particle release and does not require CAML for this effect

Milnacipran for the management of fibromyalgia syndrome

Fibromyalgia syndrome (FMS) is a widespread pain condition associated with fatigue, cognitive dysfunction, sleep disturbance, depression, anxiety, and stiffness. Milnacipran is one of three medications currently approved by the Food and Drug Administration in the United States for the management of adult FMS patients. This review is the second in a three-part series reviewing each of the approved FMS drugs and serves as a primer on the use of milnacipran in FMS treatment including information on pharmacology, pharmacokinetics, safety and tolerability. Milnacipran is a mixed serotonin and norepinephrine reuptake inhibitor thought to improve FMS symptoms by increasing neurotransmitter levels in descending central nervous system inhibitory pathways. Milnacipran has proven efficacy in managing global FMS symptoms and pain as well as improving symptoms of fatigue and cognitive dysfunction without affecting sleep. Due to its antidepressant activity, milnacipran can also be beneficial to FMS patients with coexisting depression. However, side effects can limit milnacipran tolerability in FMS patients due to its association with headache, nausea, tachycardia, hyper- and hypotension, and increased risk for bleeding and suicidality in at-risk patients. Tolerability can be maximized by starting at low dose and slowly up-titrating if needed. As with all medications used in FMS management, milnacipran works best when used as part of an individualized treatment regimen that includes resistance and aerobic exercise, patient education and behavioral therapies

Simulation study of pressure and temperature dependence of the negative thermal expansion in Zn(CN)(2)

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Small scale vegetable oil extraction

The processes required to manufacture human food from oilseeds and nuts vary considerably, depending on the raw materials and the scale of the operation. Large-scale producers can select equipment from a range of manufacturers, but the choice for smaller manufacturers is generally limited. Small-Scale Vegetable Oil Extraction presents the options available for processing a range of common oilseeds from domestic to village scales of operation. Details are provided on the type and source of equipment, and the requirements for financial success are examined. The publication will be of interest both to entrepreneurs and NGOs considering setting up oilseed operations