Gammaretroviral pol sequences act in cis to direct polysome loading and NXF1/NXT-dependent protein production by gag-encoded RNA
BACKGROUND: All retroviruses synthesize essential proteins via alternatively spliced mRNAs. Retrovirus genera, though, exploit different mechanisms to coordinate the synthesis of proteins from alternatively spliced mRNAs. The best studied of these retroviral, post-transcriptional effectors are the trans-acting Rev protein of lentiviruses and the cis-acting constitutive transport element (CTE) of the betaretrovirus Mason-Pfizer monkey virus (MPMV). How members of the gammaretrovirus genus translate protein from unspliced RNA has not been elucidated.
RESULTS: The mechanism by which two gammaretroviruses, XMRV and MLV, synthesize the Gag polyprotein (Pr65Gag) from full-length, unspliced mRNA was investigated here. The yield of Pr65Gag from a gag-only expression plasmid was found to be at least 30-fold less than that from an otherwise isogenic gag-pol expression plasmid. A frameshift mutation disrupting the pol open reading frame within the gag-pol expression plasmid did not decrease Pr65Gag production and 398 silent nucleotide changes engineered into gag rendered Pr65Gag synthesis pol-independent. These results are consistent with pol-encoded RNA acting in cis to promote Pr65Gag translation. Two independently-acting pol fragments were identified by screening 17 pol deletion mutations. To determine the mechanism by which pol promoted Pr65Gag synthesis, gag RNA in total and cytoplasmic fractions was quantitated by northern blot and by RT-PCR. The pol sequences caused, maximally, three-fold increase in total or cytoplasmic gag mRNA. Instead, pol sequences increased gag mRNA association with polyribosomes ~100-fold, a magnitude sufficient to explain the increase in Pr65Gag translation efficiency. The MPMV CTE, an NXF1-binding element, substituted for pol in promoting Pr65Gag synthesis. A pol RNA stem-loop resembling the CTE promoted Pr65Gag synthesis. Over-expression of NXF1 and NXT, host factors that bind to the MPMV CTE, synergized with pol to promote gammaretroviral gag RNA loading onto polysomes and to increase Pr65Gag synthesis. Conversely, Gag polyprotein synthesis was decreased by NXF1 knockdown. Finally, overexpression of SRp20, a shuttling protein that binds to NXF1 and promotes NXF1 binding to RNA, also increased gag RNA loading onto polysomes and increased Pr65Gag synthesis.
CONCLUSION: These experiments demonstrate that gammaretroviral pol sequences act in cis to recruit NXF1 and SRp20 to promote polysome loading of gag RNA and, thereby license the synthesis of Pr65Gag from unspliced mRNA.
BACKGROUND: In addition to the core catalytic machinery, bacterial replicative DNA polymerases contain a Polymerase and Histidinol Phosphatase (PHP) domain whose function is not entirely understood. The PHP domains of some bacterial replicases are active metal-dependent nucleases that may play a role in proofreading. In E. coli DNA polymerase III, however, the PHP domain has lost several metal-coordinating residues and is likely to be catalytically inactive.
RESULTS: Genomic searches show that the loss of metal-coordinating residues in polymerase PHP domains is likely to have coevolved with the presence of a separate proofreading exonuclease that works with the polymerase. Although the E. coli Pol III PHP domain has lost metal-coordinating residues, the structure of the domain has been conserved to a remarkable degree when compared to that of metal-binding PHP domains. This is demonstrated by our ability to restore metal binding with only three point mutations, as confirmed by the metal-bound crystal structure of this mutant determined at 2.9 A resolution. We also show that Pol III, a large multi-domain protein, unfolds cooperatively and that mutations in the degenerate metal-binding site of the PHP domain decrease the overall stability of Pol III and reduce its activity.
CONCLUSIONS: While the presence of a PHP domain in replicative bacterial polymerases is strictly conserved, its ability to coordinate metals and to perform proofreading exonuclease activity is not, suggesting additional non-enzymatic roles for the domain. Our results show that the PHP domain is a major structural element in Pol III and its integrity modulates both the stability and activity of the polymerase.
Background and Purpose: Hodgkin lymphoma (HL) treatment has evolved to reduce or avoid radiotherapy (RT) dose and volume and minimize the potential for late effects. Some older adolescents are treated on adult protocols. The purpose of this study is to examine the protocol assignment of older adolescents and its impact on radiation dose to relevant thoracic structures.
Materials and Methods: Cooperative group data were reviewed and 12 adolescents were randomly selected from a pediatric HL protocol. Treatment plans were generated per one pediatric and two adult protocols. Dose volume histograms for heart, lung, and breast allowed comparison of radiation dose to these sites across these three protocols.
Results: A total of 15.2% of adolescents were treated on adult HL protocols and received significantly higher radiation dosage to heart and lung compared to pediatric HL protocols. Adolescents treated on either pediatric or adult protocols received similar RT dose to breast.
Conclusion: Older adolescents treated on adult HL protocols received higher RT dose to thoracic structures except breast. Level of nodal involvement may impact overall RT dose to breast. The impact of varying field design and RT dose on survival, local, and late effects needs further study for this vulnerable age group.
For centuries the criminal justice system has struggled to define the methodology of and the justifications for social control of sexual behavior that does not conform to community mores. This poster compares and contrasts the historical and contemporary attempts in the United States, Canada, Belgium, the United Kingdom, and Germany to address the risk created by individuals who engage in behaviors broadly characterized as sexually deviant. Where available, we consider the rationale for sentencing, and the earliest attempts to bring “treatment” into the criminal dispositional formula for sexual based prosecution. We also consider the impact that the choice of societal response has on risk assessment and evaluation in the various systems, including where available, the assessment and commitment of juvenile offenders. The current practice of civil commitment for a person deemed to be a sexually violent predator (SVP) is discussed highlighting the U.S. Supreme Court decision in Kansas v. Hendricks. This practice will then be compared and contrasted with the approach of designating an offender as a Dangerous Offender (DO) or a Long-Term Offender (LTO) under the criminal law. We also highlight sex offender registries where applicable. This poster is intended as an overview of the law as it exists, and not as a defense or a critique of any specific model.
Association of Demographic Factors and Comorbid Diagnoses with Crime Typein an Arrest Cohort with Schizophrenia and/or Related Psychosis
The implications of the interface between the criminal justice system and individuals with schizophrenia persist despite decades of research on criminalization and risk of arrest. Research exploring the broader construct of criminality has predominantly focused on individuals with severe mental illness as a collective. This study diverges from others by examining diagnoses comorbid with schizophrenia and related psychoses and their relationships with risk of arrest across a spectrum of criminal categories ranging in severity.
Helping law enforcement, health care, and social service communities work together to develop a seamless continuum of care for persons with serious mental illness who are charged with non-serious crimes.
A recent survey of state statutes for outpatient commitment (Torrey and Kaplan, 1995) indicates that while thirty-five states and the District of Columbia have laws permitting outpatient commitment, Massachusetts is not one of them. Rather, Massachusetts uses a competency-based, substituted-decision-making model for the involuntary administration of medication in the community. To appreciate the Massachusetts model, it is important to understand how this court-ordered involuntary outpatient treatment fits into the overall scheme of outpatient commitment and how it is structured.
A review of involuntary outpatient treatment (IOT) literature indicates that it is prudent to distinguish between outpatient commitment, conditional release, and conservatorship-guardianship (Torrey and Kaplan, 1995). Two states whose IOT is based on the guardianship process and is described in the literature are California and New Mexico. Lamb and Weinberger (1992, 1993) have discussed California’s use of guardians for the gravely disabled psychiatric outpatient, and Schneider-Braus (1986) has presented a single case report from New Mexico.
Temporal Patterns of Arrest in a Cohort of Adults Receiving Mental Health Services: The Massachusetts Mental Health / Criminal Justice Cohort Study
Criminal Justice Involvement among Clientele is a Major Concern for State Mental Health Agencies. Mental health and criminal justice systems provide services at various points along the interface of these systems to reduce offending and re-offending, including:
- Diversion programs
- Mental Health Courts
Little information about scope of offending to guide service development. This study provides data on the prevalence, type and temporal patterns of arrest for a large sample of adults followed for roughly 9.5 years.
Serious Mental Illness and Chronic Crimial Justice Involvement: Findings from The Massachusetts Mental Healthy / Criminal Justice Cohort Study
Presents findings from 1990 through 2000 from The Massachusetts Mental Health/Criminal Justice Cohort Study.
Institutional Review Boards (IRBs) are the primary organizations designed to protect research subjects from harm and assure that they participate voluntarily. At the same time, many researchers feel that they intrude into the research process without making research safer.
• Identify which issues about applications are the focus of IRB attention; e.g., the scientific validity of a protocol, issues of risk, informed consent
• Clarify how, if at all, the occupants of different roles (chair, community member, attorney, scientific expert, etc.) differ in their discussion of applications
• Describe how IRB members identify problems in applications; what information resources they use and how they use them
• Identify how IRBs organize the work of application review through the use of staff, pre-meeting review, and formatl meetings
Presentation on how to receive a high survey response rate using the Dillon Method, Survey Monkey and various contact steps.
Regional differences in the reporting of mental health symptoms among juvenile justice (JJ) involved youth were examined using data from the 70,423 youths in the MAYSI-2 national norm study (Vincent et al., 2008). The percentage of youth scoring above Caution on MAYSI-2 scales was examined by race/ethnicity (white vs. minorities) and sex. Regional differences were assessed using Cochran’s Mantel-Haenszel (CMH) analyses. White youth were more likely to score above caution on all clinical scales except Depressed-Anxious. An interesting gender and race/ethnicity effect emerged such that White male youth in the Northeast and Midwest were more likely than Minority youth to score above caution; whereas White female youth in the West were more likely than Minority youth to score above caution.
This poster reports on factors that influenced the rapid adoption and implementation of the Massachusetts Youth Screening Instrument-version 2 (MAYSI-2) and the perceived consequences of routine MAYSI-2 mental health screening. Semi-structured interviews and focus groups were conducted with administrators, managers, and front-line staff in juvenile detention centers in three states (n=19). Results will allow us to better inform juvenile justice facilities regarding the conditions under which screening can more often result in increases in mental health services to youth entering the system and help guide future efforts to provide technology to juvenile justice programs in the interest of youths.
Presents a study regarding gender and race differences in juvenile mental health symptoms, using the MAYSI-2 National Norm Study.
Trajectories of Offending from Childhood to Early Adulthood in Girls With and Without Mental Health System Involvement
Criminology literature is overwhelmingly based in studies of males, though studies of gender differences or of females are rapidly accumulating. Rates of psychiatric disorder are typically higher in females involved with justice systems compared to males. However, the juvenile or criminal justice involvement of girls in mental health systems, or with serious mental health conditions is greatly understudied. Identifying their arrest risk onset, peak, and offset provides practitioners information about when to intervene and with whom. The goal of the present study is to describe within-individual longitudinal arrest patterns from ages 8-24 in this population, and determine whether their arrest patterns differ from general offender females in ways that have practice implications.
Methods: Using statewide administrative data from the Massachusetts Department of Mental Health (DMH) and Massachusetts’ juvenile and criminal courts, a database was constructed that contained juvenile and criminal arrest histories to age 25 for females born 1976-79. DMH females were adolescent service users (n=738), Non-DMH females had no DMH database records (n=34,436). Massachusetts Census 2000 provided the size of the general female population. Developmental trajectory modeling was used to group individuals’ patterns of offending over time (trajectories) into “clusters” of those whose trajectories are similar, and describe trajectories. Trajectory comparison methods minimized the greater Non-DMH cohort size.
Results: DMH females were far more likely to be arrested by age 25 than Non-DMH females (46% vs. 22%) and to be arrested at multiple ages (28% vs. 7%). Analyses revealed eight justice system trajectories among those with multiple ages of arrest. Trajectories varied on level of involvement and timing of onset/offset/peaks. Non-DMH females comprised at least 93% of each trajectory cluster, though several clusters showed significant over- or under-representation of DMH females.
Conclusions: Concern about justice system involvement of female youths in intensive MH services is justified. Among girls with multiple ages with arrest, differences in criminal careers between the mental health and non mental health system users was minimal. Implications of trajectory findings for timing and type of intervention will be presented.
Knowledge Exchange as Program Evaluation: The Family Networks Implementation Study as a Case Example
The Family Networks is a comprehensive system transformation initiative to redesign and integrate traditional categorical services across the Commonwealth into local service systems for children, youth, and families served by the child welfare system. The Family Networks Implementation Study (FNIS), a partnership between MA/DCF and UMMS, is a study of the process of implementing systems change, and provides a case example of knowledge exchange
Clubhouses, built on the principles of recovery and psychiatric rehabilitation, have nottraditionally identified and responded to the parenting related goals of club members.This poster presents findings for the conditions necessary to support Clubhouse members in their roles as parents.
The Family Options Project: Implementing an Innovative Intervention for Parents with Mental Illnesses and Their Families
The Family Options Project reflects a productive partnership between researchers and providers at the UMMS Center for Mental Health Services Research and Employment Options, Inc., Marlborough, MA The Family Options Intervention is an evidence-informed psychiatric rehabilitation intervention, developed and tested within the context of a community-based agency setting.
Approximately 760,000 emerging adults use outpatient psychotherapy in the U.S. each year (Olfson et al.2002). Emerging adults are 1.6-7.9 times more likely to drop out of mental health treatment than Fully mature adults (Edlund et.al., 2002; Olfson Et al., 2002).This Study compared temporal patterns of attendance and non attendance between emerging and mature adults.
We developed Motivational Enhancement Therapy (MET) treatment for emerging adults.