The Viewing Reaction Time as a Diagnostic Tool of Pedohebephilia in the Dunkelfeld.

Diagnosing pedohebephilia is fraught with obstacles given the tabooed nature of this sexual preference. The viewing reaction time effect (VRT) provides a non-intrusive indirect measure of sexual interest in minors. In forensic populations, the ability of the difference between the latencies while viewing child and adult sexual stimuli (VRT index) to discern child sexual offenders from a range of control groups has been ascertained meta-analytically. Given that the effect has been studied almost exclusively in forensic samples, its dependence or independence on prior overt (deviant) sexual behavior remains unclear. The present study sought to examine the relationship of prior sexual and non-sexual behaviors with the VRT in a sample of 282 self-referring, help-seeking men with and without pedohebephilia with and without a history of prior child sexual offenses (CSO) or a use of child sexual abuse materials (CSAM) recruited outside a forensic context. We found that (1) the clinical diagnosis of pedohebephilia but not prior CSO or CSAM showed a significant association with the VRT index; (2) the discriminatory ability of the VRT index did not differ significantly between samples with and without a history of prior overt sexual behavior with children; (3) the VRT index correlated positively with a behavioral marker of pedohebephilia in a subsample of individuals with prior judicially detected or undetected overt sexual behavior with children; and (4) in the same subsample, the VRT index correlated positively with markers of sexual interests in minors or hypersexuality but not of antisociality. Equivalence testing failed to refute a potential effect of prior sexual behavior on the VRT index. Our study showed that the VRT may provide an unintrusive diagnostic tool for pedohebephilia. The effect of prior overt sexual behavior with children needs further examination.

Balancing selection in Pattern Recognition Receptor signalling pathways is associated with gene function and pleiotropy in a wild rodent.

Pathogen-mediated balancing selection is commonly considered to play an important role in the maintenance of genetic diversity, in particular in immune genes. However, the factors that may influence which immune genes are the targets of such selection are largely unknown. To address this, here we focus on Pattern Recognition Receptor (PRR) signalling pathways, which play a key role in innate immunity. We used whole-genome resequencing data from a population of bank voles (Myodes glareolus) to test for associations between balancing selection, pleiotropy and gene function in a set of 123 PRR signalling pathway genes. To investigate the effect of gene function, we compared genes encoding (a) receptors for microbial ligands versus downstream signalling proteins, and (b) receptors recognizing components of microbial cell walls, flagella and capsids versus receptors recognizing features of microbial nucleic acids. Analyses based on the nucleotide diversity of full coding sequences showed that balancing selection primarily targeted receptor genes with a low degree of pleiotropy. Moreover, genes encoding receptors recognizing components of microbial cell walls etc. were more important targets of balancing selection than receptors recognizing nucleic acids. Tests for localized signatures of balancing selection in coding and noncoding sequences showed that such signatures were mostly located in introns, and more evenly distributed among different functional categories of PRR pathway genes. The finding that signatures of balancing selection in full coding sequences primarily occur in receptor genes, in particular those encoding receptors for components of microbial cell walls etc., is consistent with the idea that coevolution between hosts and pathogens is an important cause of balancing selection on immune genes.

Misuse of Child Sexual Abuse Images: Treatment Course of a Self-Identified Pedophilic Pastor.

The recurrent use of child sexual abuse images (CSAI) for sexual arousal is considered a valid indicator for the existence of a pedophilic preference, which in turn represents a significant risk factor for committing contact offenses against children. Treatment programs for (potentially) sexually delinquent men endeavor to reduce the risk to offend by addressing relevant risk factors and emphasizing functional and socially acceptable alternative behaviors. Users of CSAI are a particular target group for prevention services and it has been suggested that the primary treatment focus should be on sexual self-regulation deficits.

Is Emotional Congruence With Children Associated With Sexual Offending in Pedophiles and Hebephiles From the Community?

Although emotional congruence with children (ECWC) is a risk factor for sexual offending against children, its conceptual validity has hardly been researched. This study aims to explore the construct of ECWC by evaluating the factor structure of the Child Identification Scale (CIS-R) and its relation to facets of sexual preference and child sexual abuse behaviors. It was hypothesized that the measure comprises consistent subscales that are differently associated with aspects of sexual preference and sexual offending against children. CIS-R data of a sample of 217 adult male pedophiles from the community were used for an exploratory principal component analysis (PCA). Group comparisons and a multinomial logistic regression analysis were conducted after including a non-pedophilic control group of 22 adult men. PCA revealed a three-factor solution for the CIS-R accounting for 30% of variance. Group comparisons found differences in overall scores and the factor labeled "Attachment to Children" between subgroups of sexual age and gender preference, but not between contact, online, and non-offenders. The regression analysis showed a pedophile sexual preference and the interaction between a hebephile sexual age preference and the factor "Attachment to Children" being associated with past offending behavior. The results indicate a wish to attach to children as core feature of the CIS-R measure assessing ECWC. It is discussed whether this is an inherent feature of pedophilia or rather an independent aspect being differently distinct in pedophiles.

The German Dunkelfeld project: a pilot study to prevent child sexual abuse and the use of child abusive images.

INTRODUCTION: Sexual interest toward prepubescents and pubescents (pedophilia and hebephilia) constitutes a major risk factor for child sexual abuse (CSA) and viewing of child abusive images, i.e., child pornography offenses (CPO). Most child sexual exploitation involving CSA and CPO are undetected and unprosecuted in the "Dunkelfeld" (German: "dark field"). AIM: This study assesses a treatment program to enhance behavioral control and reduce associated dynamic risk factors (DRF) in self-motivated pedophiles/hebephiles in the Dunkelfeld. METHODS: Between 2005 and 2011, 319 undetected help-seeking pedophiles and hebephiles expressed interest in taking part in an anonymous and confidential 1-year-treatment program using broad cognitive behavioral methodology in the Prevention Project Dunkelfeld. Therapy was assessed using nonrandomized waiting list control design (n=53 treated group [TG]; n=22 untreated control group [CG]). MAIN OUTCOME MEASURES: Self-reported pre-/posttreatment DRF changes were assessed and compared with CG. Offending behavior characteristics were also assessed via self-reporting. RESULTS: No pre-/postassessment changes occurred in the control group. Emotional deficits and offense-supportive cognitions decreased in the TG; posttherapy sexual self-regulation increased. Treatment-related changes were distributed unequally across offender groups. None of the offending behavior reported for the TG was identified as such by the legal authorities. However, five of 25 CSA offenders and 29 of 32 CPO offenders reported ongoing behaviors under therapy. CONCLUSIONS: Therapy for pedophiles/hebephiles in the Dunkelfeld can alter child sexual offending DRF and reduce-related behaviors. Unidentified, unlawful child sexual exploitative behaviors are more prevalent in this population than in officially reported recidivism. Further research into factors predictive of problematic sexual behaviors in the Dunkelfeld is warranted.

Covalent immunoglobulin labeling through a photoactivable synthetic Z domain.

Traditionally, labeling of antibodies has been performed by covalent conjugation to amine or carboxyl groups. These methods are efficient but suffer from nonspecificity, since all free and available amine/carboxyl groups have the possibility to react. This drawback may lead to uncontrolled levels and locations of the labeling. Hence, the labeled molecules might behave differently and, possibly, the binding site of the antibody will also be affected. In this project, we have developed a highly stringent method for labeling of antibodies by utilizing an immunoglobulin-binding domain from protein A, the Z domain. Domain Z has been synthesized with an amino acid analogue, benzoylphenylalanine, capable of forming covalent attachment to other amino acids upon UV-exposure. This feature has been used for directed labeling of immunoglobulins and subsequent use of these in different assays.

Affinity-based entrapment of the HER2 receptor in the endoplasmic reticulum using an affibody molecule.

Interference with the export of cell surface receptors can be performed through co-expression of specific affinity molecules designed for entrapment in the endoplasmic reticulum during the export process. We describe the investigation of a small (6 kDa) non-immunoglobulin-based HER2 receptor binding affibody molecule (Z(HER2:00477)), for use in affinity mediated entrapment of the HER2 receptor in the ER. Constructs encoding Z(HER2:00477) or a control affibody protein, with or without ER-retention peptide extensions (KDEL), were expressed in the HER2 over-expressing cell line SKOV-3. Intracellular expression of the full-length affibody constructs could be confirmed by probing cell extracts by Western blotting. Confocal immunofluorescence microscopy experiments showed extensive co-localization of the HER2 receptor and Z(HER2:00477)-KDEL in the ER, whereas the use of a KDEL-extended control affibody molecule resulted in distinct and separate signals from cell surface-localized HER2 receptor and ER-localized affibody protein. This indicated a capability of the Z(HER2:00477)-KDEL fusion protein to functionally interfere with the export process of HER2 receptor in a specific manner. Using flow cytometry and cell proliferation analyses, it could be shown that expression of the Z(HER2:00477)-KDEL fusion construct in the SKOV-3 cell line resulted both in a marked reduction in cell surface level of HER2 receptors and that the cell population doubling time was significantly increased. Expression of the Z(HER2:00477)-KDEL fusion protein in additional cell lines of different origin and with different expression levels of endogenous HER2 receptor compared to SKOV-3, also resulted in depletion of the cell surface levels of HER2 receptor. This indicated upon a general ability of the Z(HER2:00477)-KDEL fusion protein to functionally interfere with the export process of HER2.

Antibodies biotinylated using a synthetic Z-domain from protein A provide stringent in situ protein detection.

Antibody-based protein profiling on a global scale using immunohistochemistry constitutes an emerging strategy for mapping of the human proteome, which is crucial for an increased understanding of biological processes in the cell. Immunohistochemistry is often performed indirectly using secondary antibodies for detection, with the benefit of signal amplification. Direct immunohistochemistry instead brings the advantage of multiplexing; however, it requires labeling of the primary antibody. Many antibody-labeling kits do not specifically target IgG and may therefore cause labeling of stabilizing proteins present in the antibody solution. A new conjugation method has been developed that utilizes a modified Z-domain of protein A (ZBPA) to specifically target the Fc part of antibodies. The aim of the present study was to compare the ZBPA conjugation method and a commercially available labeling kit, Lightning-Link, for in situ protein detection. Fourteen antibodies were biotinylated with each method and stained using immunohistochemistry. For all antibodies tested, ZBPA biotinylation resulted in distinct immunoreactivity without off-target staining, regardless of the presence of stabilizing proteins in the buffer, whereas the majority of the Lightning-Link biotinylated antibodies displayed a characteristic pattern of nonspecific staining. We conclude that biotinylated ZBPA domain provides a stringent method for antibody biotinylation, advantageous for in situ protein detection in tissues.

Androgen deprivation therapy of self-identifying, help-seeking pedophiles in the Dunkelfeld.

Androgen deprivation therapy (ADT) is considered an effective strategy in sexual offender treatment. However, the evidence base concerning its effects on sexual arousal control is limited. Past research has focused almost exclusively on men in forensic contexts. The present retrospective observational study provided data on ADT in a sample of self-identifying, help-seeking pedohebephilic men applying for a one-year group therapy program. Factors possibly influencing the readiness to take up or discontinue ADT were presented. Effects of a combination of ADT and group psychotherapy program on changes in paraphilic sexual behavior and associated psychological factors were examined. The proportion of men having taken up ADT was rather small (n=15). Greater awareness of potentially risky situations to commit child sexual offenses and self-rated uncontrollability of sexual urges were identified as characterizing men resorting to ADT. Additionally, these men were initially more open to include medical treatment. Examination of the effects of ADT and psychotherapy was limited to a sample of six men providing complete data sets. Descriptive data demonstrated a reduction of paraphilic sexual behaviors, an increase of risk-awareness and self-efficacy, and a decrease of offense-supportive cognitions and self-esteem. The present study underlined the importance of careful education and monitoring of self-identifying, help-seeking pedohebephilic patients interested in ADT concerning the effects and side effects of the treatment in a clinical context.

High-throughput protein production--lessons from scaling up from 10 to 288 recombinant proteins per week.

The demand for high-throughput recombinant protein production has markedly increased with the increased activity in the field of proteomics. Within the Human Protein Atlas project recombinantly produced human protein fragments are used for antibody production. Here we describe how the protein expression and purification protocol has been optimized in the project to allow for handling of nearly 300 different proteins per week. The number of manual handling steps has been significantly reduced (from 18 to 9) and the protein purification has been completely automated.