Background: Patients with a rectal foreign body (RFB) are still a rare entity in general surgery departments but with an increasing incidence over the last years. This case is sometimes difficult to treat, and due to a lack of standardized treatment options, the aim of the study was to present our clinical experiences with the diagnostic and therapeutic approach to RFBs and a review of the currently available literature. Materials and methods: Data were collected retrospectively from the patient's records of 20 patients who were treated due to an RFB between 2006 and 2016. Patient's demographics, circumstances of insertion, inserted objects, clinical presentation, laboratory and imaging results, as well as surgical treatment and duration of hospital stay were analyzed. Additionally, a review of the literature was performed with the search items "rectal foreign body" and "surgical therapy". Because many publications were just case reports, we did not perform a meta-analysis or a systematic review. Results: Twenty-two cases in 20 patients (80% male) presented to the emergency room. The mean age was 38.5 ± 13.7 years. In 68.2% of the cases, the cause of RFB was due to sexual preferences. The following objects were inserted: six dildos, three vibrators, two bottles, one glass, one deodorant, one apple, one fever thermometer,

Background: Congenital neutropenia (CN) is a group of rare inborn genetic disorders of hematopoiesis including a variety of different gene mutations and different patterns of inheritance. Independent of the underlying genetic subtypes, CN patients benefit from G-CSF maintenance treatment, which improved the life expectancy and quality of life significantly. The majority of CN patients documented by the SCNIR Europe respond well to G-CSF treatment and achieve and maintain neutrophil response with an anticipated ANC of greater 1000/µL with a median G-CSF dose of 4.77 µg/kg/day. However, the G-CSF dose used is very heterogeneous and dose increase is dependent on the decision of each treating physician. Here we report on the incidence, clinical characteristics and outcome of patients receiving G-CSF doses of 20µg/kg/d or higher. Methods: In the European database of the SCNIR we have identified 420 CN patients with different genetic subtypes (including SDS and GSD1b) who are treated with G-CSF. Non-response is defined as median neutrophil counts below 500/µL upon G-CSF doses of 20µg/kg/day or higher. Partial responders have median ANCs of 500 to 999/µL with these G-CSF doses or severe infections despite ANCs of greater 1000/µL. Eighty of the 420 pts received G-CSF doses of 20µg/kg/d or higher. Results: According to the above described response criteria we identified 26 non- responders (6.2%), 18 partial responders (4.3%) and 36 patients reaching ANCs above 1000 with G-CSF doses of 20µg/kg/d or higher (32 patients receiving 20 to 39µg/kg/d; 4 patients receiving more than 50µg/kg/d) out of the 420 CN patients treated with G-CSF. Although patients with different genotypes receive long-term G-CSF treatment, only ELANE or JAGN1 mutations were identified in both non- and partial responders. Analysing ELANE patients separately, 9.3% were non-responders and 5.3 % partial responders to G-CSF. While gender ratio male/female is almost even in the CN cohort (1.08 including 6 males with x-linked TAZ mutations) as well as in the ELANE subgroup (0.87), the gender ratio for ELANE non-responders is 0.2 and 0.4 in ELANE partial responders. In 50% of the identified non- and partial responders the genotype is still unclassified. In 3 patients who were not included in the analysis G-CSF non-response was due to a homozygous CSF3R mutation. These patients respond well to GM-CSF but not to G-CSF. Conclusions: Non-response or partial response to high doses of G-CSF treatment is correlated with distinct genotypes (ELANE, JAGN1, SRP54 and so far unclassified). However, requirement of high G-CSF doses (20µg/kg/d or higher) does not per se indicate non-response, since a substantial number of patients reach sufficient ANCs with these G-CSF doses, especially in patients with ELANE and HAX1 genotype. Treating physicians often do not increase G-CSF doses to >20µg/kg/d to rule out response to high doses. Patients with CSF3R homozygous mutations cannot respond to G-CSF, but respond well to GM-CSF, while ELANE or JAGN1 patients do not respond to GM-CSF. While patients with HAX1 mutations may require high G-CSF doses, but then respond well, patients with SBDS or other mutations (e.g. G6PC3, CXCR4, GFI-1, TAZ etc.) did not require high G-CSF doses for a longer period or were non-responsive to G-CSF treatment. Our data support the dosing recommendation of the SCNIR not to stop G-CSF at 20µg/kg/day, but to increase further, since a substantial number of patients are still responsive. Disclosures No relevant conflicts of interest to declare.

Background: Primary Autoimmune Neutropenia (AIN) is the most frequent type of neutropenia in children with a prevalence of 1/100,000 between infancy and 10 years of age. Primary AIN is caused by anti-neutrophil antibodies binding to neutrophil-specific antigens, resulting in a decrease of circulating neutrophils in the blood, but normal numbers of mature neutrophils in the bone marrow. Typically, primary AIN is present from infancy on until spontaneous remission in early childhood, when anti-neutrophil antibodies disappear. Patients with primary AIN show severe to moderate neutropenia but only rarely suffer from serious infections. Patients remain in the registry for follow up after normalization of blood counts to evaluate late secondary events. Aims: Here we describe the cohort of AIN pts with positive anti-neutrophil antibody testing documented by the European Branch of the SCNIR. We analyzed the course of neutropenia, the frequency of G-CSF treatment for AIN, the incidence of severe bacterial infections and administration of AB prophylaxis. Methods: We identified 102 primary AIN patients within the neutropenia cohort documented by the European Branch of the SCNIR since 1994. We classified primary AIN by positive anti-neutrophil antibody testing (95 pts) or severe neutropenia in peripheral blood with normal bone marrow morphology in patients with age under 5 years (7 pts). Results: Primary AIN has been identified in 102 (61 female; 41 male) pts. The median age of the cohort is 5.18 years (range 1.37-22.71 years), with 630.28 pt years under observation. Median age at diagnosis was 12.07 months (range 0.9-70 months). All pts are currently alive, 40 patients already resolved from primary AIN at a median age of 3.02 years (range 0.83-9.08 years). Median follow-up time after neutropenia had resolved was 2.25 years (range 0-9.27 years). Sixteen of 102 pts (15.7%) received intermittent G-CSF treatment with a median dose of 4.5 µg/kg/day compared to 4.77 µg/kg/day for the congenital neutropenia cohort of the SCNIR Europe. Analysis of infections (tab.1) showed less minor and severe infections comparing to congenital neutropenia (CN) pts. Life-threatening infections like liver abscesses were not seen in primary AIN patients but in 1.8% of CN pts. Twelve AIN pts (11.7%) have received antibiotic prophylaxis for prevention of infection, 6 pts intermittent and 6 pts continuously. However, antibiotic prophylaxis was usually stopped before termination of AIN. Due to the milder course of infections most AIN pts were able to go to Kindergarten and to live a normal life. In 3 pts additional auto-immune related diseases were identified (autoimmune thrombocytopenia, allergic colitis and Kawasaki syndrome) during AIN. Sixteen of 102 AIN patients received genetic analysis, with no mutation being detected. In addition to the 102 AIN pts we identified another 31 CN pts who have initially been classified as AIN due to positive anti-neutrophil antibodies, but who were later genetically confirmed as CN (15 ELANE+, 8 HAX1+, 2 SBDS+, 2 CXCR4+, 2 CSF3R+, 1 G6PC3+). This proportion of pts showed more and more severe infections compared to primary AIN. Genetic testing has been performed in these pts due to ongoing infections and prolonged neutropenia until school age. Conclusions: Pts suffering from primary AIN present with severe to moderate neutropenia. A minority of pts might require G-CSF treatment on demand/interventionally due to antibiotic resistant infections, but long-term G-CSF treatment is regularly not indicated. Primary AIN is a self-limiting condition and in most pts neutropenia resolves until early childhood. Accumulation of secondary diagnoses like autoimmune related diseases, though postulated, has not been confirmed for AIN pts by our data. In AIN pts with severe infections, or prolonged neutropenia CN should be ruled out by genetic analysis and/or bone marrow morphology. Registries are needed to document long-term data on primary AIN pts to analyse potential additional features of primary AIN, possibly other accompanying autoimmune diseases. Disclosures No relevant conflicts of interest to declare.

The t(8;21)(q22;q22) found in about 8% of de novo AML fuses the Runx1 (or AML-1) gene to the ETO (or MTG8) gene. The leukemia-initiating chimeric RUNX1/ETO fusion protein is believed to repress transcription of RUNX1 target genes by ETO-mediated recruitment of transcriptional repressors. Although several studies analyzing short-term depletion of RUNX1/ETO reveal both widespread epigenetic reprogramming and altered protein expression of several signaling molecules depending on RUNX1/ETO (e.g. Ptasinska et al. 2012, Spirin et al. 2014) the molecular mechanisms involved are still not completely understood. We analyzed RUNX1/ETO dependent expression of the myeloid transcription factor PU.1 in t(8;21) Kasumi cells after lentiviral transduction with control or RUNX1/ETO specific shRNAs (shCtrl and shRE). PU.1 is a well known target of RUNX1/ETO being inactivated by the chimeric protein. PU.1 expression was analyzed by immunoblotting of cellular lysates from cells transduced to > 99%. RUNX1/ETO is depleted upon shRE expression, and shRE cells exhibit reduced proliferation, less apoptotic cell death and reduced clonal growth in limiting dilution assays as compared to controls. Around day 7 after transduction expression of the full length ~37 kD PU.1 protein is specifically reduced in shRE cells, and a shorter variant of about 22 kD is increasingly expressed starting at day 4. A 20.7 kD PU.1 variant can be predicted from an alternative transcript missing exon 1 encoding the start ATG for full length PU.1. The alternative transcript encodes an ATG in frame in exon 3 with identical 3´-terminal PU.1 sequences. We cloned several PU.1 cDNAs from Kasumi cells and identified and confirmed the alternative transcript missing exon 1. In vitro transcription/translation of cloned PU.1 transcripts generated a protein with C-terminal PU.1 epitopes of the predicted size. However, using PU.1 isoform-specific quantitative real-time RT-PCR we did not find differential changes in isoform-specific mRNA expression in shRE as compared to shCtrl cells with the full length variant always being most abundant. PU.1 expression can be modulated by an antisense RNA regulated by a shared cis-regulatory element (Ebralize et al. 2008) but its expression was similar in shRE and shCtrl cells as determined by quantitative RT-PCR. Since these data suggest that differential protein expression is not due to differential abundance of specific mRNAs we analyzed expression and phosphorylation status of eIF2-α. eIF2-α is involved in translation regulation during cellular stress response by redirecting ribosomes to alternative open reading frames (ORFs) in a phosphorylation dependent manner (for review Young and Wek 2016). eIF2-α phosphorylation increases at ~ 4 days after transduction in shRE but not in shCtrl cells. Interestingly, protein expression of several proteins such as FOXO3, STAT3, STAT5 and C/EBPα is reduced in shRE as compared to shCtrl cells without significant changes in the respective mRNA levels. Furthermore, expression of the RUNX1/ETO targets PU.1, C/EBPα and BCL2 initially increases before final reduction. These data suggest that RUNX1/ETO modulates mRNA translation in addition to its well-known transcriptional and epigenetic effects. Furthermore, RUNX1/ETO targets identified by loss-of function studies may characterize individual candidates as part of a more general response to translational regulation. Disclosures No relevant conflicts of interest to declare.