Proteasome activities decrease during dexamethasone-induced apoptosis of thymocytes

The induction of apoptosis in thymocytes by the glucocorticoid dexamethasone was used as a model system to investigate whether there are changes in 20 S and 26 S proteasome activities during apoptosis. We observed that thymocytes contain high concentrations of proteasomes and that following treatment with dexamethasone, cell extracts showed a decrease in proteasome chymotrypsin-like activity which correlated with the degree of apoptosis observed. The decrease in chymotrypsin-like activity of 20 S and 26S proteasomes was still apparent after these complexes had been partially puri®ed from apoptotic thymocyte extracts and was therefore not due to competition resulting from a general increase in protein turnover. The trypsin-like and peptidylglutamylpeptide hydrolase activities of proteasome complexes were also observed to decrease during apoptosis, but these decreases were reversed by the inhibition of apoptosis by the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp(OMe)-¯uoromethylketone. However, the chymotrypsin-like activity of proteasomes decreased further in the presence of the apoptosis inhibitor. Val-Ala-Asp-¯uoromethylketone was found to inhibit the chymotrypsin- and trypsin-like activity of 26 S proteasomes in .itro. The decrease in proteasome activities in apoptosis did not appear to be due to a decrease in the concentration of total cellular proteasomes. Thus, the early decreases in 20 S and 26 S proteasome activities during apoptosis appear to be due to a down-regulation of their proteolytic activities and not to a decrease in their protein concentration. These data suggest that proteasomes may be responsible, in thymocytes, for the turnover of a protein that functions as a positive regulator of apoptosis.

Catalytic properties of 26 S and 20 S proteasomes and radiolabeling of MB1, LMP7, and C7 subunits associated with trypsin-like and chymotrypsin-like activities*

20 and 26 S proteasomes were isolated from rat liver. The procedure developed for the 26 S proteasome resulted in greatly improved yields compared with previously published methods. A comparison of the kinetic properties of 20 and 26 S proteasomes showed significant differences in the kinetic characteristics with certain substrates and differences in the effects of a protein substrate on peptidase activity. Observed differences in the kinetics of peptidylglutamyl peptide hydrolase activity suggest that the 26 S complex cannot undergo the conformational changes of 20 S proteasomes at high concentrations of the substrate benzyloxycarbonyl (Z) -Leu-Leu-Glu-b-naphthylamide. Various inhibitors that differentially affect the trypsin-like and chymotrypsin-like activities have been identified. Ala-Ala-Phe-chloromethyl (CH2Cl) inhibits chymotrypsin-like activity assayed with succinyl (Suc) -Leu-Leu-Val-Tyr-AMC, but surprisingly not hydrolysis of Ala-Ala-Phe-7-amido-4-methylcoumarin (AMC). Tyr-Gly-Arg-CH2Cl inhibits Suc-Leu-Leu-Val-Tyr-AMC hydrolysis as well as trypsinlike activity measured with t-butoxycarbonyl (Boc) -Leu-Ser-Thr-Arg-AMC, while Z-Phe-Gly-Tyr-diazomethyl (CHN2) was found to inhibit only the two chymotrypsin- like activities. Radiolabeled forms of peptidyl chloromethane and peptidyl diazomethane inhibitors, [3H]acetyl-Ala-Ala-Phe-CH2Cl, [3H]acetyland radioiodinated Tyr-Gly-Arg-CH2Cl, and Z-Phe-Gly- Tyr-(125I-CHN2), have been used to identify catalytic components associated with each of the three peptidase activities. In each case, incorporation of the label could be blocked by prior treatment of the proteasomes with known active site-directed inhibitors, calpain inhibitor 1 or 3,4-dichloroisocoumarin. Subunits of labeled proteasomes were separated either by reverse phase-HPLC and SDS-polyacrylamide gel electrophoresis or by twodimensional polyacrylamide gel electrophoresis followed by autoradiography/fluorography and immunoblotting with subunit-specific antibodies. In each case, label was found to be incorporated into subunits C7, MB1, and LMP7 but in different relative amounts depending on the inhibitor used, consistent with the observed effects on the different peptidase activities. The results strongly suggest a relationship between trypsin-like activity and chymotrypsin-like activity. They also help to relate the different subunits of the complex to the assayed multicatalytic endopeptidase activities

Good secrets, bad secrets : disclosure in children’s picture books about sexual child abuse

Well over 50 picture books have been published for children on the topic of sexual child abuse (Lampert & Walsh, 2010) many with the aim of teaching their very young readers how to tell the difference between good and bad secrets. This paper looks at three recent picture books for how they focus on disclosure as an end point.

Mother-child communication about sexual abuse prevention

Two hundred and twelve Australian mothers completed an online survey examining features of mother-child communication about child sexual abuse prevention. Two-thirds (67.5%) of respondents had discussed child sexual abuse prevention with their children. Proportions of mothers talking with their children about child sexual abuse prevention varied according to age range (highest for mothers with children aged 5-12 years) and only child status (lowest for mothers of only children). The number of topics discussed with their children differed according to child gender (greater number of topics discussed by mothers with both girls and boys) and age range (greater number of topics discussed by mothers with children aged 5-12 years). These findings provide new insights into mother-child communication about child sexual abuse prevention.

Design process excludes users : the co-creation activities between user and designer

User needs are a fundamental element of design. If the design process does not properly reflect user needs, the design will be severely compromised. Therefore, it is worthwhile to investigate how the user is, and user needs are, understood in the design process. In this article, three accepted linear process models for web site and interactive media design are reviewed in terms of the designer and user participation. The article then proposes a user-evolving collaborative design process which is built on co-creation activities between designer and user. Co-creation activities across the entire design process structurally and ontologically reposition the users, and user needs, centrally, which allows the designers to holistically approach to the user needs through building a partnership with the users. Co-creation creates an equal evolving participatory process between user and designer towards sharing values and knowledge and creating new domains of collective creativity.

Victims Speak: Comparing Child Sexual Abusers and Their Victims Accounts of Offence Circumstances

This article presents findings from a longitudinal study. The research aimed to explore the effectiveness of a treatment program for offenders which lasted for three years. The research design was structured around the program with interviews and psychometric testing undertaken at key points in time with the same group of respondents. View all notes that sought to evaluate a treatment program for child sexual abusers. A triangulated methodological approach was adopted drawing upon quantitative and qualitative methodological techniques. The focus here is upon one element of this research. 2 2The quantitative element of this research will be published shortly but is referred to in the following reports Davidson 2000, 2003 [research funded by the National Probation Service]. Psychometric testing was undertaken over a four-year period with the men attending the treatment program to explore shifts in the extent of denial, blame attribution, and victim empathy over time. Offender cognitive distortions, general health, and self-esteem were also explored via psychometric testing. An interview-administered survey was undertaken with all sex offenders registered with the Probation Service (those on probation and in custody) in order to gather demographic data, and 117 of 150 offenders responded. View all notes Ninety-one in-depth interviews were conducted over a four-year period with a small, nonrandom sample of twenty-one male offenders who had been convicted of sexual offenses against children. All of the men were subject to probation orders with a psychiatric condition (Criminal Justice Act, 1991). One of the aims of this element of the research was to explore the extent to which evidence of denial could be found in offenders’ accounts of offense circumstance and also to explore the extent to which offenders minimized the nature and extent of abuse perpetrated. Offenders’ accounts of offense circumstances were compared to victim statements, and stark differences emerge. These findings have considerable implications for treatment practice with sex offenders, where victims’ perceptions could be used to directly confront offender denial and minimization.

Child Victims of Sexual Abuse: Children's Experience of the Investigative Process in the Criminal Justice System

This paper is a selected review of research on issues surrounding the investigation of intra-familial child sexual abuse for children aged eight and above, in the criminal justice system. Particular attention is paid to features of the investigative interview in relation to the child's level of understanding, ability to report and likely emotional response when the proceedings take place. Best practice by police and social care agencies involves establishing valid and reliable information from children while attending to their developmental level and emotional state. The review aims to distil principles optimising this process from both the investigative judicial perspective and the child's focus, as well as from the inter-agency perspective and information sharing. Recommendations are made for improving the interview process based on research and methods from a range of disciplines and to optimise information recording in a format easily shared between agencies. Updated and ongoing training procedures are key to successful practice with training shared across police and social work agencies. The focus of this review is informed by preliminary findings from pilot research in progress on behalf of the Metropolitan Police Child Abuse Investigation Command.

Protecting Vulnerable Young People in Cyberspace from Sexual Abuse: Raising Awareness and Responding Globally

This paper explores the danger that young people may be exposed to when using the Internet. The first part sets out the context by considering new developments in UK and international legislation and then explores educational moves to protect children. Focus is upon findings from research undertaken on behalf of the London Metropolitan Police Service in evaluating the Safer Surfing programme designed to enable young people’s safe Internet use. In the final part of this paper it is argued that more must however be done internationally both to protect children online and to curb the growing trade in indecent child images.

Characteristics of the Internet for Criminal Child Sexual Abuse by Online Groomers

This paper presents a number of characteristics of the Internet that makes it attractive to online groomers. Relevant Internet characteristics include disconnected personal communication, mediating technology, universality, network externalities, distribution channel, time moderator, low‐cost standard, electronic double, electronic double manipulation, information asymmetry, infinite virtual capacity, independence in time and space, cyberspace, and dynamic social network. Potential sex offenders join virtual communities, where they meet other persons who have the same interest. A virtual community provides an online meeting place where people with similar interests can communicate and find useful information. Communication between members may be via email, bulletin boards, online chat, web‐based conferencing or other computer‐based media.

Separation/Divorce Sexual Assault: The Current State of Social Scientific Knowledge

Many contend that the logical solution to woman abuse in marriage/cohabitation is for women to exit through legal separation, divorce, or other means. However, a growing body of empirical work shows that separation or divorce does not necessarily solve the problem of woman abuse. For example, in addition to experiencing lethal or nonlethal forms of physical violence and psychological abuse, many women who try to leave, or who have left their male partners, are sexually assaulted. The main objective of this paper is to critically review the extant empirical and theoretical work on separation/divorcesexual assault. Suggestions for future research and theorizing are also provided.