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Transcription factors (TFs) regulate target genes by specific interactions with DNA sequences. Detecting and understanding these interactions at the molecular level is of fundamental importance in biological and clinical contexts. Crosslinking mass spectrometry is a powerful tool to assist the structure prediction of protein complexes but has been limited to the study of protein-protein and protein-RNA interactions. Here, we present a femtosecond laser-induced crosslinking mass spectrometry (fliX-MS) workflow, which allows the mapping of protein-DNA contacts at single nucleotide and up to single amino acid resolution. Applied to recombinant histone octamers, NF1, and TBP in complex with DNA, our method is highly specific for the mapping of DNA binding domains. Identified crosslinks are in close agreement with previous biochemical data on DNA binding and mostly fit known complex structures. Applying fliX-MS to cells identifies several bona fide crosslinks on DNA binding domains, paving the way for future large scale ex vivo experiments.
Subject terms: DNA-binding proteins, Proteomics, Mass spectrometry, Transcription, Structural biology
Atomic-resolution mapping of transcription factor-DNA interactions by femtosecond laser crosslinking and mass spectrometry
Alexander Reim,1 Roland Ackermann,2 Jofre Font-Mateu,3 Robert Kammel,2 Miguel Beato,3,4 Stefan Nolte,2,5 Matthias Mann,1 Christoph Russmann,corresponding author6,7 and Michael Wierercorresponding author1
Standardized methods for assessing attachment disorders are scarce but needed for research and practice.
In the current study, several assessments for attachment disorder symptoms are used within a German sample of foster children after being exposed to neglect and maltreatment in their biological families. The symptoms were assessed with four established assessment methods based on both parents’ report and behavioral observation: The Rating for Infant Stranger Engagement, the Stranger at the Door, the Disturbances of Attachment Interview and the Reactive Attachment Disorder Questionnaire.
The foster care sample showed symptoms of both the inhibited and the disinhibited attachment disorder. The degree of symptoms is comparable to previous findings. The results of the different tools investigating the disinhibited type of attachment disorder are correlated to each other, but do not overlap.
Although all approaches are based on the clinical criteria of the DSM-IV, the assessments do not coincide. Each tool provides a different point of view on the symptoms, so a multi methodical approach for assessing attachment disorder symptoms should be implemented. Furthermore, the inhibited and the disinhibited symptoms represent separate categories, as reflected in the DSM-5, requiring separate assessment.
Reactive attachment disorder (RAD), Disinhibited social engagement disorder (DSED), Diagnosis, Foster care, Assessment
Assessment of attachment disorder symptoms in foster children: comparing diagnostic assessment tools
Josephine D. Kliewer-Neumann,corresponding author1 Janin Zimmermann,2 Ina Bovenschen,2 Sandra Gabler,2 Katrin Lang,2 Gottfried Spangler,2 and Katja Nowacki1
Diagnosis and treatment of breast cancer have changed profoundly over the past 25 years. The outcome improved dramatically and was well quantified for early stage breast cancer (EBC). However, progress in the treatment of metastatic disease has been less convincingly demonstrated. We have studied survival data of patients with metastatic breast cancer (MBC) from a large academic cancer center over a period of 20 years.
Data from 1033 consecutive MBC patients who were treated at the Department of Medical Oncology of the West German Cancer Center from January 1990 to December 2009 were retrospectively analyzed for overall survival (OS) and risk factors. Patients were grouped in 5-year cohorts, and survival parameters of each cohort were compared before and after adjustment for risk factors.
Overall survival of patients with MBC treated at specialized center has significantly improved from 1990 to 2010 (hazard ratio 0.7, 95%CI 0.58-0.84). The increments in OS have become less profound over time (median OS 1990-1994: 24.2 months, 1995-1999: 29.6 months, 2000-2004: 36.5 months, 2005-2009: 37.8 months).
Survival of patients with MBC has improved between 1990 and 2004, but less from 2005 to 2009. Either this suggests an unnoticed shift in the patient population, or a lesser impact of therapeutic innovations introduced in the most recent period.
Long-term survival, Metastatic breast cancer, New drugs
Improved survival in metastatic breast cancer: results from a 20-year study involving 1033 women treated at a single comprehensive cancer center
Anja Welt,corresponding author1 Simon Bogner,1 Marina Arendt,2,4 Josef Kossow,1,5 Antonia Huffziger,1,6 Christian Pohlkamp,1,7 Heike Steiniger,1,8 Ute Becker,1,9 Ferras Alashkar,1,10 Marzena Kohl,1,11 Marcel Wiesweg,1 Heike Richly,1 Jorg Hense,1 Max E. Scheulen,1,11 Martin Schuler,1,3 Siegfried Seeber,1,12 and Mitra Tewes1