Applicability of tandem affinity purification MudPIT to pathway proteomics in yeast

A combined multidimensional chromatography-mass spectrometry approach known as "MudPIT" enables rapid identification of proteins that interact with a tagged bait while bypassing some of the problems associated with analysis of polypeptides excised from SDS-polyacrylamide gels. However, the reproducibility, success rate, and applicability of MudPIT to the rapid characterization of dozens of proteins have not been reported. We show here that MudPIT reproducibly identified bona fide partners for budding yeast Gcn5p. Additionally, we successfully applied MudPIT to rapidly screen through a collection of tagged polypeptides to identify new protein interactions. Twenty-five proteins involved in transcription and progression through mitosis were modified with a new tandem affinity purification (TAP) tag. TAP-MudPIT analysis of 22 yeast strains that expressed these tagged proteins uncovered known or likely interacting partners for 21 of the baits, a figure that compares favorably with traditional approaches. The proteins identified here comprised 102 previously known and 279 potential physical interactions. Even for the intensively studied Swi2p/Snf2p, the catalytic subunit of the Swi/Snf chromatin remodeling complex, our analysis uncovered a new interacting protein, Rtt102p. Reciprocal tagging and TAP-MudPIT analysis of Rtt102p revealed subunits of both the Swi/Snf and RSC complexes, identifying Rtt102p as a common interactor with, and possible integral component of, these chromatin remodeling machines. Our experience indicates it is feasible for an investigator working with a single ion trap instrument in a conventional molecular/cellular biology laboratory to carry out proteomic characterization of a pathway, organelle, or process (i.e. "pathway proteomics") by systematic application of TAP-MudPIT

Australian women's prediagnostic values and influencing sociodemographic variables relating to treatment choices for early breast cancer treatment

Women are often asked by their doctors to choose their preferred treatment for early breast cancer. Evidence shows that many women are distressed and confused about how to make this treatment decision and frequently seek help from nurses. Very little is known about women's value-centred decision-making in relation to selecting treatment for breast cancer and for nurses it is difficult to know how to assist these women with this process. In this study, 377 women participated prior to undergoing routine mammography screening and the data were collected using the Pre-Decision Portfolio Questionnaire (PDPQ) by Pierce 1. The partipants identified that expected treatment outcomes were the most important factor in choosing early breast cancer treatment. The majority reported that it was very important that a treatment would reduce the chances the cancer would return (95.6%), increase the length of their life (82.1%) and lead them to being healthy (80.4%). In addition, the participants indicated that it was important, or very important, that the emotional consequences of the treatment did "not make you depressed" (88.6%) or "sad" (90.4%) and should "keep you from worrying" (97%) and "give you peace of mind" (98.6%). Other factors, such as treatment's side effects, were identified as less important. Age, employment, education and having a family history of breast cancer were found to be significant influencing variables on the values of the participants. It was concluded that assessing and understanding the treatment values of women can help nurses focus on areas of importance to the woman and lead to informed decision-making when they are choosing treatment for early breast cancer

Spinning Interdisciplinary Service-Learning Webs: A Secondary Education Approach

Springboards: The ideas presented are not meant to be comprehensive. Placing this much information on one page restrains the number of ideas that can be given. They are meant to spark creative minds. Each teacher knows his/her students, community. and curriculum best. Making connections: Each and every curricular area does not need to participate for successful interdisciplinary learning to occur. Choosing your action: We have presented an example of each type of action: (direct, indirect, and advocacy). Only one type of action is required to make this a service-learning project along with preparation and reflection All School activities: When reading the Aging and Hunger and Homelessness webs it becomes obvious that an all school project is the focus. In the Aging web , references to holding a Senior Prom is a theme throughout. The same is true of the Hunger and Homelessness web where holding a Hunger Banquet becomes a school wide project. MSPAP and Service-learning: Service-learning and MSPAP are natural fit. Both incorporate real-life learning, problem-solving, critical thinking, and the opportunity to apply skills. Goals of this project: It is hoped that these webs accomplish 2 goals: 1- to increase understanding of service learning and its contributions to application of knowledge.2- to encourage curricular webbing that helps increase overall learning and understanding

Learning What Works: A Structured Evaluation of Two Tools That Help Groups Share Research Libraries

F1000 Workspace and Mendeley (with an institutional license that enables shared groups) are tools that facilitate building shared libraries of relevant content for groups such as research labs, hospital residents, and paper co-authors. The purpose of this structured evaluation is to identify the pros and cons of each tool for these users and to help librarians make informed recommendations.

Uptake, Accuracy, Safety, and Linkage into Care over Two Years of Promoting Annual Self-Testing for HIV in Blantyre, Malawi: A Community-Based Prospective Study

Background Home-based HIV testing and counselling (HTC) achieves high uptake, but is difficult and expensive to implement and sustain. We investigated a novel alternative based on HIV self-testing (HIVST). The aim was to evaluate the uptake of testing, accuracy, linkage into care, and health outcomes when highly convenient and flexible but supported access to HIVST kits was provided to a well-defined and closely monitored population. Methods and Findings Following enumeration of 14 neighbourhoods in urban Blantyre, Malawi, trained resident volunteer-counsellors offered oral HIVST kits (OraQuick ADVANCE Rapid HIV-1/2 Antibody Test) to adult (≥16 y old) residents (n = 16,660) and reported community events, with all deaths investigated by verbal autopsy. Written and demonstrated instructions, pre- and post-test counselling, and facilitated HIV care assessment were provided, with a request to return kits and a self-completed questionnaire. Accuracy, residency, and a study-imposed requirement to limit HIVST to one test per year were monitored by home visits in a systematic quality assurance (QA) sample. Overall, 14,004 (crude uptake 83.8%, revised to 76.5% to account for population turnover) residents self-tested during months 1–12, with adolescents (16–19 y) most likely to test. 10,614/14,004 (75.8%) participants shared results with volunteer-counsellors. Of 1,257 (11.8%) HIV-positive participants, 26.0% were already on antiretroviral therapy, and 524 (linkage 56.3%) newly accessed care with a median CD4 count of 250 cells/μl (interquartile range 159–426). HIVST uptake in months 13–24 was more rapid (70.9% uptake by 6 mo), with fewer (7.3%, 95% CI 6.8%–7.8%) positive participants. Being “forced to test”, usually by a main partner, was reported by 2.9% (95% CI 2.6%–3.2%) of 10,017 questionnaire respondents in months 1–12, but satisfaction with HIVST (94.4%) remained high. No HIVST-related partner violence or suicides were reported. HIVST and repeat HTC results agreed in 1,639/1,649 systematically selected (1 in 20) QA participants (99.4%), giving a sensitivity of 93.6% (95% CI 88.2%–97.0%) and a specificity of 99.9% (95% CI 99.6%–100%). Key limitations included use of aggregate data to report uptake of HIVST and being unable to adjust for population turnover. Conclusions Community-based HIVST achieved high coverage in two successive years and was safe, accurate, and acceptable. Proactive HIVST strategies, supported and monitored by communities, could substantially complement existing approaches to providing early HIV diagnosis and periodic repeat testing to adolescents and adults in high-HIV settings

BH3 domains of BH3-only proteins differentially regulate Bax-mediated mitochondrial membrane permeabilization both directly and indirectly

activate Bax directly and can only act indirectly to reliev

Palliative Care for People with Advanced Liver Disease: A Feasibility Trial of a Supportive Care Liver Nurse Specialist

Background:Liver disease is an increasing cause of death worldwide but palliative care is largely absent for these patients.Aim:We conducted a feasibility trial of a complex intervention delivered by a supportive care liver nurse specialist to improve care coordination, anticipatory care planning and quality of life for people with advanced liver disease and their carers.Design:Patients received a 6-month intervention (alongside usual care) from a specially trained liver nurse specialist. The nurse supported patients/carers to live as well as possible with the condition and acted as a resource to facilitate care by community professionals. A mixed-method evaluation was conducted. Case note analysis and questionnaires examined resource use, care planning processes and quality-of-life outcomes over time. Interviews with patients, carers and professionals explored acceptability, effectiveness, feasibility and the intervention.Setting/participants:Patients with advanced liver disease who had an unplanned hospital admission with decompensated cirrhosis were recruited from an inpatient liver unit. The intervention was delivered to patients once they had returned home.Results:We recruited 47 patients, 27 family carers and 13 case-linked professionals. The intervention was acceptable to all participants. They welcomed access to additional expert advice, support and continuity of care. The intervention greatly increased the number of electronic summary care plans shared by primary care and hospitals. The Palliative care Outcome Scale and EuroQol-5D-5L questionnaire were suitable outcome measurement tools.Conclusion:This nurse-led intervention proved acceptable and feasible. We have refined the recruitment processes and outcome measures for a future randomised controlled trial

Perceived Social Support Network and Achievement : Mediation by Motivational Beliefs and Moderation by Gender

Research has shown that perceived social support (PSS) (from parents and teachers) influences achievement. However, little is known about how this relationship operates. This study examines the multiple mediational effects of students’ motivational beliefs in relationship to the association between PSS and mathematics achievement. The sample included the African countries that participated in the TIMSS 2011 (Ghana, Botswana, South Africa, Morocco, and Tunisia). A bootstrap analysis indicated a unique pattern of the role of motivational beliefs in mediating the relationships between PSS and achievement. Moreover, gender was found to moderate the indirect effect in some countries. The findings indicate that total mediation effect of students’ motivational belief on the relationship between PSS and achievement is “culture-fair but not culture-free”Research has shown that perceived social support (PSS) (from parents and teachers) influences achievement. However, little is known about how this relationship operates. This study examines the multiple mediational effects of students’ motivational beliefs in relationship to the association between PSS and mathematics achievement. The sample included the African countries that participated in the TIMSS 2011 (Ghana, Botswana, South Africa, Morocco, and Tunisia). A bootstrap analysis indicated a unique pattern of the role of motivational beliefs in mediating the relationships between PSS and achievement. Moreover, gender was found to moderate the indirect effect in some countries. The findings indicate that total mediation effect of students’ motivational belief on the relationship between PSS and achievement is “culture-fair but not culture-free”.Peer reviewe