Role of Primary Care in Suicide Prevention During the COVID-19 Pandemic.

Primary care providers have an important role in suicide prevention, knowing that among people who die by suicide, 83% have visited a primary care provider in the prior year, and 50% have visited that provider within 30 days of their death, rather than a psychiatrist. The psychosocial impact of the coronavirus disease 2019 pandemic poses increased risk for suicide and other mental health disorders for months and years ahead. This article focuses on screening tools, identification of the potentially suicidal patient in the primary care setting, and a specific focus on suicide prevention during widespread, devastating events, such as a pandemic.

Mitochondrial membrane potential identifies cells with high recombinant protein productivity.

Development of cell lines for biotherapeutic protein production requires screening large numbers of clones to identify and isolate high producing ones. As such, stable cell line generation is a time- and resource-intensive process. There is an increasing need to enhance the selection efficiency of high-yielding clonal cell lines for cell line development projects by using high throughput screening of live cells for markers predictive of productivity. Single cell deposition by fluorescence activated cell sorting (FACS) is a commonly performed method for cloning to generate cell lines derived from a single recombinant cell. We have developed a novel strategy to identify higher productivity cells at the FACS step by leveraging a simple viable cell staining method that detects mitochondrial membrane potential (Ψm), a key indicator of cellular metabolic activity. We chose a dual-emission dye (Mito-ID, Enzo Life Sciences) that fluoresces green and orange in living cells with the intensity of the orange fluorescence being dependent on the cells Ψm status. Using available clonal cell lines with known productivity, or stable transfectant pools, we evaluated Ψm of cell populations with Mito-ID dye. We determined that the intensity of the Ψm fluorescent signal correlates with the known fed-batch titers of the producer clones, and that cell sorting based on an optimal Ψm staining intensity selectively enriches for higher producing clones from nonclonal transfectant pools. These clones are phenotypically stable for recombinant protein production. Furthermore, the strategy has been successfully applied to identifying higher producing cell lines for a range of antibody molecular formats. Using this method, we can combine an enriching step with the cloning step for high producers, thereby saving time and resources in cell line development.

High-throughput screening of antibody-expressing CHO clones using an automated shaken deep-well system.

Biopharmaceutical protein manufacturing requires the highest producing cell lines to satisfy current multiple grams per liter requirements. Screening more clones increases the probability of identifying the high producers within the pool of available transfectant candidate cell lines. For the predominant industry mammalian host cell line, Chinese hamster ovary (CHO), traditional static-batch culture screening does not correlate with the suspension fed-batch culture used in manufacturing, and thus has little predictive utility. Small scale fed-batch screens in suspension culture correlate better with bioreactor processes but a limited number of clones can be screened manually. Scaled-down systems, such as shaken deep well plates, combined with automated liquid handling, offer a way for a limited number of scientists to screen many clones. A statistical analysis determined that 384 is the optimal number of clones to screen, with a 99% probability that six clones in the 95th percentile for productivity are included in the screen. To screen 384 clones efficiently by the predictive method of suspension fed-batch, the authors developed a shaken deep-well plate culturing platform, with an automated liquid handling system integrating cell counting and protein titering instruments. Critical factors allowing deep-well suspension culture to correlate with shake flask culture were agitation speed and culture volume. Using our automated system, one scientist can screen five times more clones than by manual fed-batch shake-flask or shaken culture tube screens and can identify cell lines for some therapeutic protein projects with production levels greater than 6 g/L. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1460-1471, 2018.

Measuring the aggregation of CHO cells prior to single cell cloning allows a more accurate determination of the probability of clonality.

The manufacturing process for biotherapeutics is closely regulated by the Food and Drug Administration (FDA), European Medicines Agency (EMA) and other regulatory agencies worldwide. To ensure consistency of the product of a manufacturing cell line, International Committee on Harmonization guidelines (Q5D, 1997) state that the cell substrate should be derived from a single cell progenitor, i.e., clonal.Cell lines in suspension culture may naturally revert to cell adhesion in the form of doublets, triplets and higher order structures of clustered cells. We can show evidence of a single colony from limiting dilution cloning or in semi-solid media, but we cannot determine the number of cells from which the colony originated. To address this, we have used the ViCELL® XR (Beckman Coulter, High Wycombe, UK) cell viability analyzer to determine the proportion of clusters of two or more cells in a sample of the cell suspension immediately prior to cloning. Here, we show data to define the accuracy of the ViCELL for characterizing a cell suspension and summarize the statistical model combining two or more rounds of cloning to derive the probability of clonality. The resulting statistical model is applied to cloning in semi-solid medium, but could equally be applied to a limiting dilution cloning process. We also describe approaches to reduce cell clusters to generate a cell line with a high probability of clonality from a CHO host lineage. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:593-601, 2018.

Cognitive behavioral therapy, self-efficacy, and depression in persons with chronic pain.

Chronic pain is a complex and often disabling condition compounded by depression and poor self-efficacy. The purpose of this evidence-based project was to explore the relationship of cognitive behavioral therapy (CBT)-focused groups with self-efficacy and depression in persons with chronic pain at an intensive interdisciplinary 3-week pain rehabilitation center (PRC). The project sample consisted of 138 persons admitted to a PRC and scoring ≥27 on the Center for Epidemiological Study Depression Scale (CES-D) and then completing the Pain Self-Efficacy Questionnaire (PSEQ). After completing the PRC program, including CBT-focused groups, discharge CES-D and PSEQ scores were analyzed. A comparison group of CES-D scores from 134 persons admitted to the PRC from a 9-month time period preceding the addition of the CBT-focused groups was also examined. There was a significant increase in self-efficacy after participation in the intensive pain rehabilitation program including CBT-focused groups. Patient groups both before and after introduction of CBT-focused groups showed the same rate of improvement on the depression scores, suggesting that persons who participated in CBT-focused groups improved equally compared with persons who did not participate in these groups. Ninety-three percent of the participants expressed satisfaction with the CBT groups. This evidence-based practice is well supported in the literature and can be implemented with knowledgeable staff and engaged stakeholders.

Group interpersonal and social rhythm therapy for bipolar depression.

PURPOSE: To evaluate the feasibility of 2-week interpersonal and social rhythm therapy group (IPSRT-G) for bipolar depression. DESIGN AND METHODS: Participants with bipolar depression received two individual sessions, six IPSRT-G sessions, and a 12-week telephone call. The Inventory of Depressive Symptomatology-Clinician Rated (IDS-C), Young Mania Rating Scale (YMRS), Sheehan Disability Scale (SDS), and Clinical Global Impressions-Bipolar Version (CGI-BP) were used. FINDINGS: IDS-C and SDS scores improved significantly at 12 weeks. YMRS and CGI-BP scores improved but did not reach statistical significance. PRACTICE IMPLICATIONS: The promising antidepressive response supports further study of IPSRT-G for bipolar depression.

Original research in the classroom: why do zebrafish spawn in the morning?

As part of an upper level undergraduate developmental biology course at the University of Minnesota Duluth, we developed a unit in which students carried out original research as part of a cooperative class project. Students had the opportunity to gain experience in the scientific method from experimental design all of the way through to the preparation of publication on their research that included text, figures, and tables. This kind of inquiry-based learning has been shown to have many benefits for students, including increased long-term learning and a better understanding of the process of scientific discovery. In our project, students designed experiments to explore why zebrafish typically spawn in the first few hours after the lights come on in the morning. The results of our experiments suggest that spawning still occurs when the dark-to-light transition is altered or absent. This is consistent with the work of others that demonstrates that rhythmic spawning behavior is regulated by an endogenous circadian clock. Our successes and failures carrying out original research as part of an undergraduate course should contribute to the growing approaches for using zebrafish to bring the excitement of experimental science to the classroom.

Proposed career pathway for clinical academic general dental practitioners.

The Modernising Medical Careers framework provides the opportunity for both medical specialists and general medical practitioners to follow training pathways that lead to appointments as National Health Service (NHS) consultants and to senior academic posts. Similar opportunities are available for dentists who wish to specialise. However, they are not available to dentists working in primary dental care who wish to become NHS consultants or senior academics in general dentistry. An alternative pathway is required that does not force committed primary care dentists who wish to become NHS consultants or senior academics down a path of specialisation. In this paper, the authors explore the situation in some detail and propose a career pathway with appropriate competencies for primary care dentists who aspire to become NHS consultants or senior academics. They justify why such posts should be created. The competencies have been developed using key guidelines and documents from the European Bologna Process and the Association for Dental Education in Europe, the Curriculum for UK Dental Foundation Programme Training, and the General Dental Council monospecialty curricula. It is hoped that the proposed pathway will produce highly trained generalists who will: (a) encourage and undertake research in primary dental care, where over 90% of dentistry is delivered, (b) support and lead outreach centres so that teaching and clinical cases reflect primary dental care, where students will spend their working lives post-qualification, and (c) provide a means of increasing the numbers of clinical dental academics, which have been in decline over the last 10 years.

The Henry street consortium population-based competencies for educating public health nursing students.

The Henry Street Consortium, a collaboration of nurse educators from universities and colleges and public health nurses (PHNs) from government, school, and community agencies, developed 11 population-based competencies for educating nursing students and the novice PHN. Although many organizations have developed competency lists for experts, the Consortium developed a set of competencies that clearly define expectations for the beginning PHN. The competencies are utilized by both education and practice. They guide nurse educators and PHNs in the creation of learning experiences that develop population-based knowledge and skills for baccalaureate nursing students. Public health nursing leaders use the competencies to frame their expectations and orientations for nurses who are new to public health nursing. This paper explains the meaning of each of the 11 population-based competencies and provides examples of student projects that demonstrate competency development. Strategies are suggested for nurse educators and PHNs to promote effective population-based student projects in public health agencies.

Transfection of cultured eukaryotic cells using cationic lipid reagents.

The development of high-efficiency methods for the introduction of functional genetic material into eukaryotic cells using cationic lipids has accelerated biological research in the studies of gene expression, control of cell growth, and cell lineage. Transfection mediated by cationic lipids is commonly used in industrial protein production as well as in some clinical gene therapy protocols. This unit describes how to perform transfection of adherent and suspension cells, insect cells, and RNA transfection using cationic lipid reagents.

Clinical nurse specialist influence in the conduct of research in a clinical agency.

Clinical nurse specialists (CNSs) have a unique view and understanding of patients in their environment and are a valuable resource that has not been adequately engaged in research, even when CNSs assist researchers in the conduct of research. The purpose of this article is to outline activities for supporting the conduct of research that capitalize on the clinical strengths of the CNS from an ecological framework. To illustrate the contributions of CNSs in the conduct of research, 3 clinical research studies are described. Discussion of these studies within the context of an ecological framework offers a systematic approach to describing the potential involvement of the CNS in the implementation of nursing research.

Developing an intervention to alter catastrophizing in persons with fibromyalgia.

PURPOSE: The purpose of this pilot study was to develop and evaluate a brief psychoeducational intervention to decrease pain catastrophizing, a focus on pain and its negative consequences in patients with fibromyalgia. DESIGN: A case-study design was used to design the targeted intervention. The intervention was piloted with 2 small groups of patients and family members. SAMPLE: Thirty-nine patients with fibromyalgia completed the surveys; patients/family members (N = 9) and ten interdisciplinary staff members participated in separate focus groups or e-mail surveys to provide data to develop the intervention. Two additional groups of patients and family members (N = 7) participated in the pilot intervention sessions. FINDINGS: Catastrophic thinking and associated declines in function were confirmed by survey results and focus group themes. A 2-hour session using self-efficacy theory was developed from these findings and other data sources. The patients/family members in pilot groups reported an increased knowledge of pain catastrophizing and satisfaction with the intervention. IMPLICATIONS FOR NURSING PRACTICE: A brief, psychoeducational offering that targets the reduction of catastrophizing is a feasible addition to the usual treatment protocol in a fibromyalgia treatment program and warrants further study.

Using portfolios to evaluate achievement of population-based public health nursing competencies in baccalaureate nursing students.

Public health nurses from 13 local public health agencies and nurse educators from five schools of nursing developed population-based public health nursing competencies for new graduates and novice public health nurses. Educators in one nursing program used a portfolio assignment to measure achievement of the competencies by traditional and RN to BSN students in a community health nursing course. Data were collected from surveys and focus groups to determine students' responses to the portfolio and their use of population-based public health nursing concepts. The assignment enhanced students' critical thinking skills; however, concerns about the structure and evaluation of the portfolio decreased student satisfaction. Recommendations are made for improving the portfolio format, increasing students' valuing of the portfolio, managing the tension between assessment and learning, and orienting clinical agency staff and nursing instructors.

Transfection of cultured eukaryotic cells using cationic lipid reagents.

The development of high-efficiency methods for the introduction of functional genetic material into eukaryotic cells using cationic lipids has accelerated biological research in the studies of gene expression, control of cell growth, and cell lineage. Transfection mediated by cationic lipids is commonly used in industrial protein production as well as in some clinical gene therapy protocols. Replacing our previous unit on this topic, this new version describes how to perform transfection of adherent and suspension cells, insect cells, and RNA transfection using the cationic lipid system.

A value - impact approach for regulatory decision making : An application to nuclear power

This paper presents an extended value-impact methodology which aids decision makers in ranking various alternative actions for reducing the risk associated with nuclear power reactor. It extends the state-of-the-art value-impact methodology by using the Analytic Hierarchy Process (AHP), a formalized decision making tool for ranking various altenatives based on judgment. The method has been applied to a value-impact study of the implementation of either has a vented-containment system or an alternative decay heat removal system as a means for reducing risk at the Grand Gulf nuclear power plant. A ranking of several policy actions which could reduce the economic risk of nuclear power is perfomed herein. The results of this analysis show that the method provides considerable insight to the solution of topics of interest in the decision making area of nuclear power risk management.(AU)