Identification of a Specific Telomere Terminal Transferase Activity in Tetrahymena Extracts

We have found a novel activity in Tetrahymena cell free extracts that adds tandem TTGGGG repeats onto synthetic telomere primers. The single-stranded DNA oligonucleotides (TTGGGG)4 and TGTGTGGGTGTGTGGGTGTGTGGG, consisting of the Tetrahymena and yeast telomeric sequences respectively, each functioned as primers for elongation, while (CCCCAA)4 and two nontelomeric sequence DNA oligomers did not. Efficient synthesis of the TTGGGG repeats depended only on addition of micromolar concentrations of oligomer primer, dGTP, and dTTP to the extract. The activity was sensitive to heat and proteinase K treatment. The repeat addition was independent of both endogenous Tetrahymena DNA and the endogenous alpha-type DNA polymerase; and a greater elongation activity was present during macronuclear development, when a large number of telomeres are formed and replicated, than during vegetative cell growth. We propose that the novel telomere terminal transferase is involved in the addition of telomeric repeats necessary for the replication of chromosome ends in eukaryotes

The Telomere Terminal Transferase of Tetrahymena Is a Ribonucleoprotein Enzyme with Two Kinds of Primer Specificity

We have analyzed the de novo telomere synthesis catalyzed by the enzyme telomere terminal transferase (telomerase) from Tetrahymena. Oligonucleotides representing the G-rich strand of telomeric sequences from five different organisms specifically primed the addition of TTGGGG repeats in vitro, suggesting that primer recognition may involve a DNA structure unique to these oligonucleotides. The sequence at the 3' end of the oligonucleotide primer specified the first nucleotide added in the reaction. Furthermore, the telomerase was shown to be a ribonucleoprotein complex whose RNA and protein components were both essential for activity. After extensive purification of the enzyme by a series of five different chromatographic steps, a few small low abundance RNAs copurified with the activity

The Regulated Child

This chapter explores the concept of participation as a right for children. Given that child participation has various definitions and is applied to a variety of attitudes, values and behaviours, it will adopt a case-study approach. Firstly, it will draw upon an LA-funded project that investigated the participation of vulnerable children and young people (CYP) in the integrated frontline service provision they received. Secondly, it will to examine core principles underpinning the United Nations Convention (1989: Article 12), in order to examine statutory views of participation rights. By these means, the chapter will seek to investigate CYPs’ exercise of participation rights as well as attempt to identify factors that might influence the exercise of participation. In so doing, the chapter will consider whether or not the statutory view provides the conditions for exercised rights

Learnings From a National Cyberattack Digital Disaster During the SARS-CoV-2 Pandemic in a Pediatric Emergency Medicine Department

Objective: The primary objective was to analyze the impact of the national cyberattack in May 2021 on patient flow and data quality in the Paediatric Emergency Department (ED), amid the SARS-CoV-2 (COVID-19) pandemic. Methods: A single site retrospective time series analysis was conducted of three 6-week periods: before, during, and after the cyberattack outage. Initial emergent workflows are described. Analysis includes diagnoses, demographic context, key performance indicators, and the gradual return of information technology capability on ED performance. Data quality was compared using 10 data quality dimensions. Results: Patient visits totaled 13 390. During the system outage, patient experience times decreased significantly, from a median of 188 minutes (pre-cyberattack) down to 166 minutes, most notable for the period from registration to triage, and from clinician review to discharge (excluding admitted patients). Following system restoration, most timings increased. Data quality was significantly impacted, with data imperfections noted in 19.7% of data recorded during the system outage compared to 4.7% before and 5.1% after. Conclusions: There was a reduction in patient experience time, but data quality suffered greatly. A hospital’s major emergency plan should include provisions for digital disasters that address essential data requirements and quality as well as maintaining patient flow

Structured sedation programs in the emergency department, hospital and other acute settings: protocol for systematic review of effects and events.

BACKGROUND: The use of procedural sedation outside the operating theatre has increased in hospital settings and has gained popularity among non-anesthesiologists. Sedative agents used for procedural pain, although effective, also pose significant risks to the patient if used incorrectly. There is currently no universally accepted program of education for practitioners using or introducing procedural sedation into their practice. There is emerging literature identifying structured procedural sedation programs (PSPs) as a method of ensuring a standardized level of competency among staff and reducing risks to the patient. We hypothesize that programs of education for healthcare professionals using procedural sedation outside the operating theatre are beneficial in improving patient care, safety, practitioner competence and reducing adverse event rates. METHODS: Electronic databases will be systematically searched for studies (randomized and non-randomized) examining the effectiveness of structured PSPs from 1966 to present. Database searches will be supplemented by contact with experts, reference and citation checking, and a grey literature search. No language restriction will be imposed. Screening of titles and abstracts, and data extraction will be performed by two independent reviewers. All disagreements will be resolved by discussion with an independent third party. Data analysis will be completed adhering to procedures outlined in the Cochrane Handbook of Systematic Reviews of Interventions. If the data allows, a meta-analysis will be performed. DISCUSSION: This review will cohere evidence on the effectiveness of structured PSPs on sedation events and patient outcomes within the hospital and other acute care settings. In addition, it will examine key components identified within a PSP associated with patient safety and improved patient outcomes.Trial registration: PROSPERO registration number: CRD42013003851

The Interpersonal Style and Complementarity Between Crisis Negotiators and Forensic Inpatients

Previous negotiation research has explored the interaction and communication between crisis negotiators and perpetrators. A crisis negotiator attempts to resolve a critical incident through negotiation with an individual, or group of persons in crisis. The purpose of this study was to establish the interpersonal style of crisis negotiators and complementarity of the interpersonal interaction between them and forensic inpatients. Crisis negotiators, clinical workers and students (n = 90) used the Check List of Interpersonal Transactions-Revised (CLOIT-R) to identify interpersonal style, along with eight vignettes detailing interpersonal styles. Crisis negotiators were most likely to have a friendly interpersonal style compared to the other non-trained groups. Complementarity theory was not exclusively supported as submissive individuals did not show optimistic judgments in working with dominant forensic inpatients and vice versa. Exploratory analysis revealed that dominant crisis negotiators were optimistic in working with forensic inpatients with a dominant interpersonal style. This study provides insight into the area of interpersonal complementarity of crisis negotiators and forensic inpatients. Whilst further research is required, a potential new finding was established, with significant ‘similarity’ found when dominant crisis negotiators are asked to work with dominant forensic inpatients

The extent, quality and impact of patient and public involvement in primary care research: a mixed methods study

Background: To improve the lives of patients in primary care requires the involvement of service users in primary care research. We aimed to explore the extent, quality and impact of patient and public involvement (PPI) in primary care research. Methods: We extracted information about PPI from grant applications, reports and an electronic survey of researchers of studies funded by the NIHR School for Primary Care Research (SPCR). We applied recognised quality indicators to assess the quality of PPI and assessed its impact on research. Results: We examined 200 grant applications and reports of 181 projects. PPI was evident in the development of 47 (24%) grant applications. 113 (57%) grant applications included plans for PPI during the study, mostly in study design, oversight, and dissemination. PPI during projects was reported for 83 (46%) projects, including designing study materials and managing the research. We identified inconsistencies between planned and reported PPI. PPI varied by study design, health condition and study population. Of 46 (24%) of 191 questionnaires completed, 15 reported PPI activity. Several projects showed best practice according to guidelines, in terms of having a PPI budget, supporting PPI contributors, and PPI informing recruitment issues. However few projects offered PPI contributors training, used PPI to develop information for participants about study progress, and had PPI in advising on dissemination. Beneficial impacts of PPI in designing studies and writing participant information was frequently reported. Less impact was reported on developing funding applications, managing or carrying out the research. The main cost of PPI for researchers was their time. Many researchers found it difficult to provide information about PPI activities. Our findings informed: - a new Cost and Consequences Framework for PPI in primary care research highlighting financial and non-financial costs, plus the benefits and harms of PPI - Fifteen co-produced recommendations to improve PPI in research and within the SPCR. Conclusions: The extent, quality and impact of PPI in primary care research is inconsistent across research design and topics. Pockets of good practice were identified making a positive impact on research. The new Cost and Consequences Framework may help others assess the impact of PPI

Local and Regional North Carolina Collaborations: Case Studies from the North Carolina Chapter of the American Planning Association

Carolina Planning regularly publishes a feature highlighting projects from members of the North Carolina Chapter of the American Planning Association (APA-NC). This year’s submissions range from multi-county visioning efforts to small-town transportation planning, demonstrating that planning at any scale can benefit from innovative collaboration. Includes the following case studies: Innovative Partnerships (Introduction); Opt-In Project Explores Uncharted Planning Territory in Southwestern NC; Transportation Planning in a Growing Community; Playing Around with Civic Engagement Strategies in the Triad; Simple Changes and Collaboration: NCDOT and the Town of West Jefferson Partner to Revitalize Downtow