A twin UGUA motif directs the balance between gene isoforms through CFIm and the mTORC1 signaling pathway.

Alternative polyadenylation (APA) generates mRNA isoforms and diversifies gene expression. Here we report the discovery that the mTORC1 signaling pathway balances the expression of two Trim9/TRIM9 isoforms through APA regulation in human and mouse. We showed that CFIm components, CPSF6 and NUDT21, promote the short Trim9/TRIM9 isoform (Trim9-S/TRIM9-S) expression. In addition, we identified an evolutionarily conserved twin UGUA motif, UGUAYUGUA, in TRIM9-S polyadenylation site (PAS) that is critical for its regulation by CPSF6. We found additional CPSF6-regulated PASs with similar twin UGUA motifs in human and experimentally validated the twin UGUA motif functionality in BMPR1B, MOB4, and BRD4-L. Importantly, we showed that inserting a twin UGUA motif into a heterologous PAS was sufficient to confer regulation by CPSF6 and mTORC1. Our study reveals an evolutionarily conserved mechanism to regulate gene isoform expression by mTORC1 and implicates possible gene isoform imbalance in cancer and neurological disorders with mTORC1 pathway dysregulation.

Shift in MSL1 alternative polyadenylation in response to DNA damage protects cancer cells from chemotherapeutic agent-induced apoptosis.

DNA damage reshapes the cellular transcriptome by modulating RNA transcription and processing. In cancer cells, these changes can alter the expression of genes in the immune surveillance and cell death pathways. Here, we investigate how DNA damage impacts alternative polyadenylation (APA) using the PAPERCLIP technique. We find that APA shifts are a coordinated response for hundreds of genes to DNA damage, and we identify PCF11 as an important contributor of DNA damage-induced APA shifts. One of these APA shifts results in upregulation of the full-length MSL1 mRNA isoform, which protects cells from DNA damage-induced apoptosis and promotes cell survival from DNA-damaging agents. Importantly, blocking MSL1 upregulation enhances cytotoxicity of chemotherapeutic agents even in the absence of p53 and overcomes chemoresistance. Our study demonstrates that characterizing adaptive APA shifts to DNA damage has therapeutic implications and reveals a link between PCF11, the MSL complex, and DNA damage-induced apoptosis.

Comprehensive profiling of mRNA polyadenylation in specific cell types in vivo by cTag-PAPERCLIP.

The ability to generate cell-type specific mRNA polyadenylation (pA) maps from complex tissues is crucial for understanding how alternative polyadenylation (APA) is regulated in individual cell types in their physiological environment under different conditions. In this chapter, we discuss cTag-PAPERCLIP, a recently developed method combining the well-established CLIP (crosslinking immunoprecipitation) technique and the Cre-lox system to achieve customized cell-type specific APA profiling from mouse tissue without cell purification or enrichment. In cTag-PAPERCLIP, selective expression of GFP-tagged poly(A) binding protein (PABP-GFP) in the desired cell type is achieved through Cre-mediated activation of a latent knock-in allele of PABP-GFP. Immunoprecipitation of PABP-GFP then allows mRNA 3' end fragments in the desired cell type to be specifically retrieved from ultraviolet (UV)-irradiated whole tissue lysate. The mRNA fragments are subsequently turned into a cDNA library to provide a comprehensive APA map and an mRNA expression profile of the chosen cell type through deep sequencing.

Healing journeys: Indigenous Men's reflections on resources and barriers to mental wellness.

Indigenous peoples in Canada and other settler colonial nations experience barriers to healing in the health care system and their communities. Drawing on four sequential sharing circles and indepth interviews with 11 Indigenous men, this article shares the stories of Indigenous men and their healing journeys with the aim of improving culturally safe support in the community. In sharing their stories, these men identified coping with colonialism, as well as trauma and grief, as barriers in their healing journey. They also described finding strength in cultural role models, fathering, as well as ceremony and connecting to the land. We discuss the implications of these findings for service provision and decolonizing community health services.

More than "petty squabbles" - Developing a contextual understanding of conflict and aggression among older women in low-income assisted living.

Dominant approaches to relational aggression among older adults tend to conceptualize the problem as a behavioral or interpersonal issue, and can inadvertently infantilize the phenomenon as 'bullying.' In this article we use a narrative approach and the conceptual lens of precarity to develop an in-depth, theoretically informed analysis of relational aggression between older women in low-income assisted living. The analysis of the narratives of tenants (and a manager) indicated that past life experiences and intersecting threats to power and identity shaped and could intensify tenants' interpretations of and reactions to others' actions and comments. Conflicts over a) unequal distributions of caring labor, b) control of social activities, and c) access to appreciation are complex and rational responses to precarious contextual conditions. Findings contribute empirically to the body of research on relational aggression among older adults, expanding this field through connecting it to critical gerontological conceptualizations of precarity. Preventing relational aggression requires increased public investment in formal social supports for older adults, challenging dominant discourses that privilege independence, and recognizing how the legacies of past disadvantage and contextual precarity (as opposed to mental illness or dementia) shape social interactions with and responses to others.

Putting responsive behaviours in place: Examining how formal and informal carers understand the actions of people with dementia.

Living alongside and working with people with dementia who yell or strike out can be very demanding and sometimes harmful. It is generally understood that such actions may be a response to the social and physical environment, yet very little attention has been paid to understanding what role the environment plays in eliciting and responding to these actions across different settings. Drawing on 27 semi-structured interviews with formal and informal carers, this article examines how carers understand the actions of people with dementia in relation to their environment. We identify individual, interpersonal, local environmental, macro-scale and temporal dynamics that influence the actions of people with dementia and carers' capacities to work with them. Drawing on relational thinking and the concept of structural violence, we argue that many of the problems and solutions associated with aggressive actions are located outside the person with dementia in broader relations and systems of care. We conclude with a discussion of how these understandings can inform developments in the systems of home and community care.

"Not there yet": Examining community support from the perspective of people with dementia and their partners in care.

People with dementia can live meaningful and engaged lives with the appropriate social and physical supports in place. There has been relatively little research, however, on the experiences and desires of people with dementia themselves as they negotiate informal and formal support in rural and small town settings. In this article, we draw on semi-structured interviews with 46 community-dwelling people with dementia and 43 partners in care in rural Ontario, Canada to examine how people with dementia relate to and within their communities as well as their perceptions of community support services. We identify the continued contributions of people with dementia to their own care and the care of others as well as common social, cultural, and organizational factors related to delayed service use and refusal to use particular services. We argue that care is "not there yet" for people in the earlier stages of dementia and that more attention needs to be paid to what people with dementia can offer their communities as well as the role of culture and gender in developing support. Our findings make an important contribution to understanding the experience of dementia in rural and small town Canada, which is relevant to rural healthcare and community support in other industrialized countries.

The dynamics of voluntarism in rural dementia care.

In this article, we contribute to the social sciences literature on voluntarism by examining the dynamics of voluntary service provision for people living with dementia in rural settings. Although volunteer-based organizations provide community support services across a range of Western countries, little attention has been directed towards understanding the organization and actions of volunteers and voluntarism in dementia care. To address this gap, we conducted a case study of Alzheimer support organizations in Ontario, Canada, using questionnaires with service providers (N=20) and semi-structured interviews with people with dementia (N=46) and partners in care (N=43). In our analysis, we identify challenges related to increasing demands for support, partner relations, reaching rural communities, a lack of early stage supports, a lack of volunteers for programs that families have requested, and loss of volunteers in programs families depended on. Moreover, we argue that the current model of voluntary sector service provision for people living with dementia is unevenly developed and potentially unsustainable.

The impact of the transcendental meditation program on government payments to physicians in Quebec: an update.

PURPOSE: To determine whether practice of the Transcendental Meditation (TM) technique can affect medical expenses. DESIGN: The evaluation was a quasi experimental, longitudinal, cost-minimization study. SETTING: Province of Quebec, Canada. SUBJECTS: This study involved 1418 Quebec health insurance enrollees who practiced the TM technique compared with 1418 subjects who were randomly selected from enrollees of the same age, sex, and region. TM subjects had chosen to begin the technique prior to learning about and choosing to enter the study. MEASURES: This 14-year, pre- and postintervention study retrospectively assessed government payments to physicians for treating the TM and comparison groups. Other medical expense data for individuals were unavailable. Data were inflation-adjusted. For each subject, least squares regression slopes were calculated to estimate pre- and postintervention annual rates of change in payments. We compared the groups' means and 1%, 5%, and 10% trimmed means (robust estimators) of the slopes. RESULTS: Before starting meditation, the yearly rate of increase in payments between groups was not significantly different (p > .17). After commencing meditation, the TM group's mean payments declined 1% to 2% annually. The comparison group's payments increased up to 11.73% annually over 6 years. There was a 13.78% mean annual difference (p = .0017). CONCLUSIONS: The results suggest that the TM technique reduced payments to physicians between 5% and 13% annually relative to comparison subjects over 6 years. Randomized studies are recommended.

The HomeSafe Pilot Program: a novel approach to injury prevention in residential construction.

Workers in the residential construction industry face unacceptably high risk of injury, disability and death. Attempts to implement comprehensive health and safety programs in this industry have met with little success. The HomeSafe Pilot Program is a novel residential construction safety program developed and sponsored by the Occupational Safety and Health Administration (OSHA) Region VIII and the Home Builders Association of Metropolitan Denver (HBA). Test subjects represent over 7475 persons employed in residential construction in the six county Denver Metro area of Colorado. The HomeSafe Pilot Program includes primary behavioral, engineering and administrative interventions to improve safe work practices in residential construction. It has some unique features of brevity, specificity and incentives not seen elsewhere in the construction industry. Its overall goal is to guide residential construction companies along a path of progressive development of comprehensive safety and health programs. The HomeSafe Pilot Program is introduced and compared to other safety and health program models developed by OSHA and the HBA. This study began in January 1997 and will continue through the millennium.

The sickle cell hemolytic transfusion reaction syndrome.

BACKGROUND: Patients with sickle cell anemia may develop serious, life-threatening hemolytic transfusion reactions (HTRs). More severe anemia may develop after the HTR than was present before transfusion, which suggests the possibility of an increased rate of hemolysis of autologous red cells. STUDY DESIGN AND METHODS: The signs and symptoms occurring during eight severe HTRs that occurred in five patients with sickle cell anemia were reviewed, as were published reports by other investigators. Calculations of red cell production and destruction incorporating known correction factors for reticulocyte maturation were performed to determine the most probable mechanism for the striking drop in hematocrit observed in several instances. RESULTS: A characteristic constellation of findings was recognized in some severe HTRs in patients with sickle cell anemia. Calculations of daily red cell production and senescence indicated that a marked drop in hematocrit occurs when erythropoiesis is suppressed in a patient with a short red cell life span and that this could account for severe posttransfusion anemia when donor red cells are hemolyzed during an HTR. CONCLUSION: A sickle cell HTR syndrome was defined. A rapid increase in the severity of anemia occurs in patients with sickle cell anemia when all donor red cells are hemolyzed during an HTR and when there is suppression of erythropoiesis, as commonly occurs as a result of transfusion or concomitant illness. Although an increased rate of hemolysis of autologous red cells may also occur, more definitive data are required to document that in these patients.

An innovative approach to reducing medical care utilization and expenditures.

In a retrospective study, we assessed the impact on medical utilization and expenditures of a multicomponent prevention program, the Maharishi Vedic Approach to Health (MVAH). We compared archival data from Blue Cross/Blue Shield Iowa for MVAH (n = 693) with statewide norms for 1985 through 1995 (n = 600,000) and with a demographically matched control group (n = 4,148) for 1990, 1991, 1994, and 1995. We found that the 4-year total medical expenditures per person in the MVAH group were 59% and 57% lower than those in the norm and control groups, respectively; the 11-year mean was 63% lower than the norm. The MVAH group had lower utilization and expenditures across all age groups and for all disease categories. Hospital admission rates in the control group were 11.4 times higher than those in the MVAH group for cardiovascular disease, 3.3 times higher for cancer, and 6.7 times higher for mental health and substance abuse. The greatest savings were seen among MVAH patients older than age 45, who had 88% fewer total patients days compared with control patients. Our results confirm previous research supporting the effectiveness of MVAH for preventing disease. Our evaluation suggests that MVAH can be safely used as a cost-effective treatment regimen in the managed care setting.

Patients' attitudes toward having been forcibly medicated.

Forced antipsychotic medication procedures are generally perceived to be clinically necessary options, albelt violations of individuals' bodies and autonomy. Previous studies have explored forcibly medicated patients' attitudes concerning these procedures, but as patients were interviewed while still in the hospital, this may have affected their responses. We interviewed consecutively forcibly medicated English-speaking acute-care inpatients after their discharge to the community. The interviews were conducted by telephone by a clinician not involved with their treatment. Of 65 such patients, 7 had already been rehospitalized, 3 could not recall the procedure, and 25 others refused the interview or were not locatable. Of the 30 who were successfully interviewed, only 47 percent had received any forced injections; the remainder had accepted oral medication under duress. Recollecting their experiences, 57 percent professed fear of side effects, 17 percent feared "addiction," and 17 percent objected to others' controlling them. Forty percent recalled feeling angry, 33 percent helpless, 23 percent fearful, 13 percent embarrassed, but 23 percent were relieved. Surprisingly, 60 percent retrospectively agreed with having been coerced, 53 percent stating they were more likely to take medication voluntarily in the future. Other forcibly medicated patients had poorer outcomes, such as rapid readmission or discharge to a state hospital: those patients may have harbored more negative feelings. However, a substantial fraction of the patients who were reached in the community appeared to support having received medication forcibly as inpatients.

Indications and immediate patient outcomes of pathology intraoperative consultations. College of American Pathologists/Centers for Disease Control and Prevention Outcomes Working Group Study.

OBJECTIVE: To evaluate the reasons (indications) for and immediate intraoperative surgical results (outcomes) associated with pathology intraoperative consultation. DESIGN: In 1992 and 1993, surgeons collaborated with pathologists in 472 voluntarily participating institutions from the United States (462), Canada (7), Australia (2), and New Zealand (1) in a study jointly sponsored by the College of American Pathologists and the Centers for Disease Control and Prevention. Pathologists selected 20 consecutive intraoperative consultations and assembled a cover letter, a checklist questionnaire, and a copy of the corresponding surgical pathology report, all of which were sent to the surgeon(s) for retrospective evaluation. PARTICIPANTS: The study was distributed to participants in the College of American Pathologists voluntary Q-Probes quality improvement and Surgical Pathology Performance Improvement programs and to Canadian and Australian hospitals with more than 200 beds. RESULTS: Evaluation of 9164 cases established the five most common indications for intraoperative consultation: (1) establish or confirm diagnosis to determine type or extent of operation (51%), (2) confirm adequacy of margins (16%), (3) confirm nature of tissue to direct sampling for immediate culture or other laboratory study (10%), (4) expedite obtaining diagnosis to inform family or patient (8%), and (5) confirm sufficient tissue submitted to secure diagnosis in permanent section (8%). The information provided by the intraoperative consultation resulted in changed surgical procedures that were either modified, terminated, or newly initiated in 47%, 30%, 6%, 9%, and 28% of cases, corresponding respectively to each of the above five common indications. Rarely cited reasons for intraoperative consultation were to expedite obtaining diagnosis for surgeon's knowledge (3%), to facilitate patient management, other professional communication or discharge planning prior to permanent section availability (3%), academic protocol (< 1%), and consultation not needed or no reason for request (< 1%). CONCLUSIONS: This multi-institutional, interdisciplinary database confirms that pathology intraoperative consultations, regardless of the initial indications, influence immediate patient care decisions, resulting in changed surgical procedures in an average of 39% of all operative cases.

The impact of the transcendental meditation program on government payments to physicians in Quebec.

PURPOSE: This study evaluated whether governmental medical payments in Quebec were affected by the Transcendental Meditation (TM) technique. DESIGN: This retrospective study used a pre- and postintervention design in which government payments for physicians' services were reviewed for 3 years before and up to 7 years after subjects started the technique. Payment data were adjusted for aging and year-specific variation (including inflation) using normative data. No separate control group was used; thus it is impossible to determine whether the changes were caused by the TM program or some other factor. SUBJECTS: A volunteer group of 677 provincial health insurance enrollees was evaluated. The subjects had chosen to practice the TM technique before they were selected to enter the study. The subjects (348 men, 329 women) had diverse occupations. Their average age was 38 years and ranged from 18 to 71 years at the start of the TM program. INTERVENTION: The TM technique of Maharishi Mahesh Yogi is a standardized procedure practiced for 15 to 20 minutes twice daily while sitting comfortably with eyes closed. SETTING: Province of Quebec, Canada. RESULTS: During the 3 years before starting the TM program, the adjusted payments to physicians for treating the subjects did not change significantly. After beginning TM practice, subjects' adjusted expenses declined significantly. The several methods used to assess the rate of decline showed estimates ranging from 5% to 7% annually. CONCLUSIONS: The results suggests that the TM technique reduces government payments to physicians. However, because of the sampling method used, the generalizability of these results to wider populations could not be evaluated.

Transfusion medicine monitoring practices. A study of the College of American Pathologists/Centers for Disease Control and Prevention Outcomes Working Group.

OBJECTIVE: To survey transfusion medicine practices in 1990, to determine the distribution of defects in the transfusion process, to examine the relationship between defects and complications, and to recommend improvements in the transfusion process. DESIGN: A mail survey that divided the transfusion process into 24 risk-prone steps and gathered defect rates on each step, along with incidence data for eight known complications of transfusions and other demographic information. SETTINGS: Hospitals, independent laboratories, and blood centers that provide transfusion medicine services. OTHER PARTICIPANTS: Respondents were 1365 participants in the College of American Pathologists 1991 Blood Bank Quality Assurance Survey. RESULTS: While processing 6.2 million units of blood and blood products, respondents reported detecting over 88,000 defects: 41% in the preanalytic phase of testing, 55% in the postanalytic phase, and only 4% in the analytic phase, the phase to which most monitoring efforts were devoted. A median of eight steps were actively monitored by survey participants overall, whereas 96 facilities sought defects in all 24 steps. CONCLUSIONS: Analysis of the data showed several monitoring steps provide similar information. Although monitoring of the transfusion process could not be linked with prevention of the complications studied, active surveillance does focus attention on defect-prone steps and allows testing of strategies to improve the transfusion process. We describe how defect detection systems may be improved.

Transfusion-associated graft-versus-host disease in immunocompetent patients: report of a fatal case associated with transfusion of blood from a second-degree relative, and a survey of predisposing factors.

A patient without evident immune deficiency who received a transfusion of blood from a second-degree family member developed fatal transfusion-associated graft-versus-host disease (TA-GVHD). The donor was homozygous for an HLA haplotype for which the recipient was heterozygous (one-way HLA match). All 39 reported cases of TA-GVHD in immunocompetent patients were reviewed to ascertain the predisposing factors and to define the indications for irradiating blood for this population. HLA typing was described in 15 cases; in 13, including seven related and six unrelated donors, a one-way HLA match was present. Thirty-one (79%) of the 39 cases were reported from Japan (and 196 other cases are cited in the Japanese literature), but a one-way HLA match among unrelated donors at HLA-A, -B, -DR loci is only approximately two to four times more likely in Japanese persons than in whites. Fresh blood (< 96 hours old) was used in 29 (94%) of the 31 cases reported from Japan and in 33 (87%) of 38 cases overall (in one case, the age of the blood used was not reported). Thus, factors that appear to predispose to TA-GVHD in immunocompetent patients are a one-way HLA match, fresh blood, and, possibly, Japanese ancestry. Irradiating cellular blood components from all blood relatives of transfusion recipients will not completely eliminate the risk of TA-GVHD.

An erythrocyte glycoprotein of apparent Mr 60,000 expresses the Sc1 and Sc2 antigens.

Immunoblotting with human anti-Sc1 and anti-Sc2 locates the Sc1 and Sc2 antigens to an erythrocyte membrane glycoprotein of apparent Mr 60,000. The antigens are destroyed by pronase, and require intact disulphide bonds for expression. A proportion of the molecules carrying the Sc1 and Sc2 antigens are associated with red cell cytoskeleton preparations. Treatment of intact cells with an Endo F preparation resulted in the loss of the Sc2 antigen but not the Sc1 antigen, suggesting that the Sc2 antigen is dependent on the presence of one or more complex N-glycans for its expression.