Paired Cohort Research Articles

CCL2-driven inflammation increases mammary gland stromal density and cancer susceptibility in a transgenic mouse model

BackgroundMacrophages play diverse roles in mammary gland development and breast cancer. CC-chemokine ligand 2 (CCL2) is an inflammatory cytokine that recruits macrophages to sites of injury. Although CCL2 has been detected in human and mouse mammary epithelium, its role in regulating mammary gland development and cancer risk has not been explored.MethodsTransgenic mice were generated wherein CCL2 is driven by the mammary epithelial cell-specific mouse mammary tumour virus 206 (MMTV) promoter. Estrous cycles were tracked in adult transgenic and non-transgenic FVB mice, and mammary glands collected at the four different stages of the cycle. Dissected mammary glands were assessed for cyclical morphological changes, proliferation and apoptosis of epithelium, macrophage abundance and collagen deposition, and mRNA encoding matrix remodelling enzymes. Another cohort of control and transgenic mice received carcinogen 7,12-Dimethylbenz(a)anthracene (DMBA) and tumour development was monitored weekly. CCL2 protein was also quantified in paired samples of human breast tissue with high and low mammographic density.ResultsOverexpression of CCL2 in the mammary epithelium resulted in an increased number of macrophages, increased density of stroma and collagen and elevated mRNA encoding matrix remodelling enzymes lysyl oxidase (LOX) and tissue inhibitor of matrix metalloproteinases (TIMP)3 compared to non-transgenic controls. Transgenic mice also exhibited increased susceptibility to development of DMBA-induced mammary tumours. In a paired sample cohort of human breast tissue, abundance of epithelial-cell-associated CCL2 was higher in breast tissue of high mammographic density compared to tissue of low mammographic density.ConclusionsConstitutive expression of CCL2 by the mouse mammary epithelium induces a state of low level chronic inflammation that increases stromal density and elevates cancer risk. We propose that CCL2-driven inflammation contributes to the increased risk of breast cancer observed in women with high mammographic density.

BEAM or BeEAM High-Dose Chemotherapy Followed By ASCT: A Single Center Comparative Analysis of Toxicity

Introduction:The BEAM (carmustine (BCNU), etoposide, aracytin and melphalan) standard conditioning regimen in autologous stem-cell transplantation is widely used since 1990 in patients with relapsed/refractory non Hodgkin's lymphoma (NHL) and Hodgkin's lymphoma (HL) who remain sensitive to salvage therapy. Recently, a phase Ib-II feasibility study using bendamustine rather than BCNU in the same indication was reported, but was not really compared to BEAM concerning safety. We report herein a safety analysis of Bendamustine-EAM (BeEAM) with a control BEAM counterpart paired cohort (2/1).Methods:We performed a case control retrospective study of patients with NHL and HL who underwent high-dose chemotherapy (HDC) with BeEAM regimen between January 2015 and December 2015. We matched each BeEAM patient with two patients having the same age, sex and number of treatment lines who underwent BEAM and ASCT between January 2008 and December 2014. No patient presented significant comorbidity. The BEAM regimen consisted in BCNU on day -6 (300mg/m2) cytarabine daily from day -6 to day -3 (200mg/m2 every 12 hours), etoposide daily from day -6 to day -3 (100mg/m2 every 12 hours) and melphalan on day -2 (140mg/m2). A similar scheme was adopted in BeEAM arm with bendamustine on day -6 and -5 (100mg/m2/d) replacing BCNU. Autologous stem cells were reinfused on day 0. Unfractionated heparin was used with Bendamustine to prevent veno-occlusive disease. Pegfilgrastim 6mg was injected subcutaneously on day 4. Febrile neutropenia was treated according to ESMO guidelines.Results:One hundred and two patients (68 BEAM and 34 BeEAM) were analyzed. Median age was 48 years in both arms. 61.8% of patients were male and 38.2% female. A median number of 4.4 x106 CD34 were reinfused in the two groups. The median time to neutrophils recovery (> 0.5 x109) was similar between the two arms (9.06 vs 8.86 days, p=0.3). Grade 3 or greater diarrhea according to Commun Terminology Criteria for Adverse Events (CTCAE v4.03) classification was significantly more frequent in BeEAM patients (44 vs 13.2%, p=0.001). Median time to hospital discharge was significantly longer in BeEAM group (23 vs 20.8 days, p=0.0047). The median loss of weight during hospitalization was significantly greater in BeEAM patients (3.3 vs 1.9 kg, p=0.014). The median number of days with fever >38°C and with intravenous antibiotics was significantly higher in BeEAM group (6.06 vs 3.38, p<0.001; 10.76 vs 8.22, p=0.0012 respectively). Five patients in BeEAM arm (14.7%) and 3 patients in BEAM (4.4%) developed grade 2-3 liver cytolysis. Four patients in BeEAM arm developed grade 1 renal toxicity. Six patients presented bacteremia (17.6%) in BeEAM versus 8 patients (11.7%) in BEAM arm. Incidence of mucositis, nausea and vomiting was similar between the two groups. Four patients in the BeEAM arm were rehospitalized for recurrent fever three to five days after discharge. No death occurred in both groups during the ASCT procedure.Conclusion:Our results based on retrospective case-control study suggest that BeEAM conditioning regimen followed by ASCT seems to be more toxic than the standard BEAM regimen. DisclosuresRibrag:gilead: Membership on an entity's Board of Directors or advisory committees; infinity: Membership on an entity's Board of Directors or advisory committees; nanostring: Membership on an entity's Board of Directors or advisory committees; pharmamar: Membership on an entity's Board of Directors or advisory committees; nanostring: Membership on an entity's Board of Directors or advisory committees; ESAI: Membership on an entity's Board of Directors or advisory committees; incyte: Membership on an entity's Board of Directors or advisory committees; incyte: Membership on an entity's Board of Directors or advisory committees; BMS: Membership on an entity's Board of Directors or advisory committees; ESAI: Membership on an entity's Board of Directors or advisory committees; BMS: Membership on an entity's Board of Directors or advisory committees; servier: Consultancy; argenX: Research Funding. Michot:Bristol-Myers Squibb: Membership on an entity's Board of Directors or advisory committees.

Perioperative Factors Influencing Readmissions of Liver Transplant Recipients with Bipolar Disorder

Introduction: Previous studies have focused on outcome of psychiatric patients with pre-transplant diagnosis of depression in orthotopic liver transplantation (OLT), but there is a dearth of data on the outcomes of patients with bipolar disorder. Specifically, questions remain regarding factors that may influence the rate of readmission following OLT. Aim: To identify factors that may influence readmission within 30 days post OLT in patients with bipolar disorder. Methods: A retrospective study was performed to include patients who had a diagnosis of bipolar disorder and underwent OLT. Data from the time interval of 01/2000 and 12/2015 was abstracted by retrospective chart review, including the patients' and donors' demographics and perioperative variables. With those factors as statistical matching indexes, the data of a paired control group was collected. The major endpoint is to calculate the 30 days readmission rate. Results: A total of 2812 patients underwent OLT at our transplant center between 01/2000 and 12/2015. Of those, 19 patients (52.9±11.5 years, 63.2% male) with a previous diagnosis of bipolar disorder were identified, and 19 patients (53.6 ±11.4 years, 63.2% male) were selected in paired cohort. Etiology of liver diseases in case group includes chronic hepatitis C infection (7 patients), hepatitis B infection (3 patients), non-alcoholic steatohepatitis (2 patients), cryptogenic cirrhosis (2 patients), and other causes. Of these 19 patients, 7 patients (37%) had hepatocellular carcinoma. In case group, 7 patients (37%) were readmitted within 30 days post-OLT, with 1 patient due to a psychological reason. Eight patients were readmitted in control group. In both groups, those patients who were readmitted have relatively higher MELD score at OLT and longer post-op ICU stay, but only statistically significant in control group. Case group had longer hospital mainly due to non-psychological complications in one patient.Figure 1Conclusion: Readmission due to psychiatric issues within 30 days following OLT was uncommon in recipients with bipolar disorder. There is no difference of post-op readmission rate between patient with or without diagnosis of bipolar disorder prior to transplant. Perioperative factors did not influence readmission or outcomes following OLT, and bipolar disorder may not be a hurdle to be considered for liver transplant. Further study with larger number of patients warranted.

The new Asian modified CKD-EPI equation leads to more accurate GFR estimation in Chinese patients with CKD.

To verify whether the new Asian modified CKD-EPI equation improved the performance of original one in determining GFR in Chinese patients with CKD. A well-designed paired cohort was set up. Measured GFR (mGFR) wasthe result of 99mTc-diethylene triamine pentaacetic acid (99mTc-DTPA) dual plasma sample clearance method. The estimated GFR (eGFR) was the result of the CKD-EPI equation (eGFR1) and the new Asian modified CKD-EPI equation (eGFR2). The comparisons were performed to evaluate the superiority of the eGFR2 in bias, accuracy, precision, concordance correlation coefficient and the slope of regression equation and measure agreement. A total of 195 patients were enrolled and analyzed. The new Asian modified CKD-EPI equation improved the performance of the original one in bias and accuracy. However, nearly identical performance was observed in the respect of precision, concordance correlation coefficient, slope of eGFR against mGFR and 95% limit of agreement. In the subgroup of GFR<60mLmin-1/1.73m2, the bias of eGFR1 was less than eGFR2 but they have comparable precision and accuracy. In the subgroup of GFR>60mLmin-1/1.73m2, eGFR2 performed better than eGFR1 in terms of bias and accuracy. The new Asian modified CKD-EPI equation can lead to more accurate GFR estimation in Chinese patients with CKD in general practice, especially in the higher GFR group.

Comparación de complicaciones asociadas con el uso de placas en 8 y tornillos canulados para la corrección de deformidades angulares

BackgroundThe epiphysiodesis and hemiepiphysiodesis for correction of angular deformities in the lower limbs has evolved over time and today the use of transient techniques is preferred like are cannulated screws and eight plates. The aim of the study was to compare the outcome of patients with angular deformities treated with cannulated screws and eight shaped plates techniques and determine if there are differences in the outcome of patients. MethodsA prospective cohort study was performed including pediatric patients treated at the Instituto de Ortopedia Infantil Roosevelt (IOIR) for angular deformities in their lower limbs from 2010 to 2012. A paired comparable cohort of patients in age, sex and diagnosis were assessed in their outcomes. For group 1 (eight shaped plate patient cohort) and group 2 (cannulated screws patient cohort) we evaluated the presence of pain, antalgic limp, edema, infection and inability to fully support lower limbs and crutches requirements. Results6-month postoperative follow-up was performed. It was found in the assessments made in the first month of monitoring the presence of pain in 10 patients for group 1 and in 2 patients for group 2 (p<0.05). We found that the antalgic limp, swelling, infection and inability to support have no statistically significant differences in the study group. DiscussionThe use of eight shaped plates is associated with the presence of increased pain in the immediate postoperative period in some patients, but this review suggests that a medium-term pain improvement is seen in this group of patients, therefor not affecting the final clinical outcome. Evidence levelIII.

Abstract 4136: Mutational analysis of paired primary and relapse diffuse large B-cell lymphoma

Abstract Recurrences of diffuse large b-cell lymphoma (DLBCL) represent a high risk of worse outcome, however knowledge on the genetic mechanism of such relapses is still lacking. In general, recurrences are considered to be outgrowths of the original neoplasm, but this concept has been challenged by rare demonstration of clonally unrelated. In our previous study we analyzed chromosomal copy number aberrations in paired primary-relapse DLBCL samples and demonstrated the existence of three different modes of tumor evolution: (i) clonally unrelated relapses, (ii) clonally related relapses via a putative common progenitor (branching evolution) and (iii) clonally related relapses with linear progression. In the current continuation study we aimed to validate our previous findings and to expand the dataset by identifying mutations in genes reported to be frequently mutated in lymphomas at primary and relapse stages of paired DLBCL samples. In addition, we compare the obtained results to the mutational profile of DLBCL that never relapsed. We designed and validated a multiplex PCR-based target-enrichment panel for the 30 most frequently affected genes in lymphomas. Target-enriched libraries were constructed using the genomic DNA of 25 matched primary-relapse pairs as well as 20 non-relapsing DLBCL samples. Sequencing was performed with the IonTorrent PGM sequencer. The data collection is currently in progress. Mutational data of the paired primary and relapse samples of DLBCL will allow us to identify DNA mutations which occur early at the primary stage and are most probably associated with disease onset. Mutations that occur at relapse will also be detected, allowing insight into genetic mechanisms that lead to DLBCL recurrence. Chromosomal aberration data available for the majority of those samples will assist the more precise interpretation of mutated allelic fractions and a holistic approach to the analysis of the affected regulatory pathways. Lack of knowledge on the genetic mechanisms of DLBCL relapse exists despite the fact that these recurrences are the main cause of morbidity and mortality in such patients. Paired sample analysis gives a unique opportunity for an insight in this field. Earlier, analyzing paired primary-relapse samples, we provided clear evidence for the existence of clonally unrelated relapses as well as different genetic patterns of DLBCL recurrence. The current study will complete the characterization of the paired DLBCL cohort at the base-pair resolution allowing identification of mutations occurring early at the onset and relapse stages of the disease. Citation Format: Darius Juskevicius, Valeria Perrina, Luca Quagliatta, Christian Ruiz, Stephan Dirnhofer, Alexandar Tzankov. Mutational analysis of paired primary and relapse diffuse large B-cell lymphoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4136. doi:10.1158/1538-7445.AM2015-4136

Visually directed vs. software-based targeted biopsy compared to transperineal template mapping biopsy in the detection of clinically significant prostate cancer

ObjectivesTargeted biopsy based on cognitive or software magnetic resonance imaging (MRI) to transrectal ultrasound registration seems to increase the detection rate of clinically significant prostate cancer as compared with standard biopsy. However, these strategies have not been directly compared against an accurate test yet. The aim of this study was to obtain pilot data on the diagnostic ability of visually directed targeted biopsy vs. software-based targeted biopsy, considering transperineal template mapping (TPM) biopsy as the reference test. Methods and materialsProspective paired cohort study included 50 consecutive men undergoing TPM with one or more visible targets detected on preoperative multiparametric MRI. Targets were contoured on the Biojet software. Patients initially underwent software-based targeted biopsies, then visually directed targeted biopsies, and finally systematic TPM. The detection rate of clinically significant disease (Gleason score ≥3+4 and/or maximum cancer core length ≥4mm) of one strategy against another was compared by 3×3 contingency tables. Secondary analyses were performed using a less stringent threshold of significance (Gleason score ≥4+3 and/or maximum cancer core length ≥6mm). ResultsMedian age was 68 (interquartile range: 63–73); median prostate-specific antigen level was 7.9ng/mL (6.4–10.2). A total of 79 targets were detected with a mean of 1.6 targets per patient. Of these, 27 (34%), 28 (35%), and 24 (31%) were scored 3, 4, and 5, respectively. At a patient level, the detection rate was 32 (64%), 34 (68%), and 38 (76%) for visually directed targeted, software-based biopsy, and TPM, respectively. Combining the 2 targeted strategies would have led to detection rate of 39 (78%). At a patient level and at a target level, software-based targeted biopsy found more clinically significant diseases than did visually directed targeted biopsy, although this was not statistically significant (22% vs. 14%, P = 0.48; 51.9% vs. 44.3%, P = 0.24). Secondary analysis showed similar results. Based on these findings, a paired cohort study enrolling at least 257 men would verify whether this difference is statistically significant. ConclusionThe diagnostic ability of software-based targeted biopsy and visually directed targeted biopsy seems almost comparable, although utility and efficiency both seem to be slightly in favor of the software-based strategy. Ongoing trials are sufficiently powered to prove or disprove these findings.

Mass gathering and globalization of respiratory pathogens during the 2013 Hajj

Every year, more than 10 million pilgrims arrive in the Kingdom of Saudi Arabia for the Hajj or Umrah. Crowding conditions lead to high rates of respiratory infections among the pilgrims, representing a significant cause of morbidity and a major cause of hospitalization. Pre- and post-Hajj nasal specimens were prospectively obtained from a paired cohort (692 pilgrims) and from nonpaired cohorts (514 arriving and 470 departing pilgrims) from 13 countries. The countries of residence included Africa (44.2%), Asia (40.2%), the United States (8.4%) and Europe (7.2%). Nasal specimens were tested for 34 respiratory pathogens using RT-PCR. A total of 80 512 PCRs were performed. The prevalence of viruses and bacteria increased, from 7.4% and 15.4% before the Hajj to 45.4% and 31.0% after the Hajj, respectively, due to the acquisition of rhinovirus, coronaviruses (229E, HKU1, OC43), influenza A H1N1, Streptococcus pneumoniae, Haemophilus influenzae and Staphylococcus aureus. We did not identify Middle East respiratory coronavirus carriage. At arrival, the prevalence of several viruses was clearly dependent on the pilgrim's country of origin. After Hajj participation, these viruses were isolated among pilgrims from all countries, with few exceptions. No significant differences were observed between paired and nonpaired cohort results. Our results strongly suggest that, given the particularly crowded conditions during the rituals, an international mass gathering such as the Hajj may contribute to the globalization of respiratory pathogens after the cross-contamination of pilgrims harbouring pathogens that easily spread among participants. Influenza and pneumococcal vaccination, face mask use and hand hygiene should be considered in the context of the Hajj.

Medulloblastoma subgroups remain stable across primary and metastatic compartments.

Medulloblastoma comprises four distinct molecular variants with distinct genetics, transcriptomes, and outcomes. Subgroup affiliation has been previously shown to remain stable at the time of recurrence, which likely reflects their distinct cells of origin. However, a therapeutically relevant question that remains unanswered is subgroup stability in the metastatic compartment. We assembled a cohort of 12-paired primary-metastatic tumors collected in the MAGIC consortium, and established their molecular subgroup affiliation by performing integrative gene expression and DNA methylation analysis. Frozen tissues were collected and profiled using Affymetrix gene expression arrays and Illumina methylation arrays. Class prediction and hierarchical clustering were performed using existing published datasets. Our molecular analysis, using consensus integrative genomic data, establishes the unequivocal maintenance of molecular subgroup affiliation in metastatic medulloblastoma. We further validated these findings by interrogating a non-overlapping cohort of 19 pairs of primary-metastatic tumors from the Burdenko Neurosurgical Institute using an orthogonal technique of immunohistochemical staining. This investigation represents the largest reported primary-metastatic paired cohort profiled to date and provides a unique opportunity to evaluate subgroup-specific molecular aberrations within the metastatic compartment. Our findings further support the hypothesis that medulloblastoma subgroups arise from distinct cells of origin, which are carried forward from ontogeny to oncology.

Outcome of fetoscopic laser ablation for twin‐to‐twin transfusion syndrome in dichorionic–triamniotic triplets compared with monochorionic–diamniotic twins

To compare outcome of dichorionic-triamniotic (DCTA) triplet gestations with that of a paired cohort of monochorionic-diamniotic (MCDA) twin gestations undergoing laser therapy for treatment of twin-to-twin transfusion syndrome (TTTS). All DCTA triplets treated at four referral centers between specified dates were included. A matched cohort group of MCDA twin gestations affected by TTTS was used for comparison. Sixteen sets of DCTA triplets treated with laser photocoagulation of placental anastomoses were compared with 32 matched sets of MCDA twins. All of the 'singleton' fetuses in the triplet gestations survived to birth and to the end of the neonatal period. There were no differences in single or double survival or the rate of non-survivors to birth and at age 28 days. A mean 3-week difference was found in the procedure-to-delivery interval in DCTA triplets vs MCDA twins (60 ± 35.8 days vs 83.6 ± 33.2 days; P = 0.029). A mean difference of 3 weeks was also found in the gestational age at delivery in DCTA triplets vs MCDA twins (28.5 ± 3.5 weeks vs 31.9 ± 5 weeks; P = 0.024). A similar post-laser fetal growth pattern in donors and recipients of both groups was noted. DCTA triplet gestations with TTTS have a similar rate of post-laser survival but deliver earlier than can be expected for twins treated with laser. These findings have potential implications for the risk of prematurity and long-term outcomes.

MP67-17 MRI–ULTRASOUND FUSION TARGETED PROSTATE BIOPSY: THE IMPORTANCE OF BIOPSYING OUTSIDE THE MRI LESION

INTRODUCTION AND OBJECTIVES: Suspicious lesions identified on multi-parametric MRI (mpMRI) may be targeted with a growing range of MRI-guided prostate biopsy techniques. It has been suggested that mpMRI could reduce the burden of prostate biopsy by limiting biopsy samples to only the areas suspicious on MRI. This study assessed the performance of mpMRI at excluding clinically significant prostate cancer outside the suspicious MRI lesion. METHODS: A paired cohort study of 104 suspicious lesions identified onmultiparametric MRI. Lesions weremarked with an arrow by an expert uro-radiologist and scored from 1 to 5 according to the European Consensus guidelines (PI-RADS). MRI-US fusion targeted biopsy of the lesion was performed using Varian brachytherapy software (Variseed 8.0.2, Varian Medical Systems). After targeting the lesion, the remaining prostate was systematically sampled with transperineal sector biopsy (min 24 max 38 cores). This technique preferentially targets the peripheral zone by sectors and any prostate cancer identified outside the lesion was located into a prostate quadrant. RESULTS: Outside the MRI lesion, clinically significant prostate cancer was identified in 39% (41/104) and there was upgrading of the Gleason score in 16% (17/104). This MRI invisible cancer was located in the same quadrant of the lesion in 9%, in an adjoining quadrant in 22% and in non-adjoining quadrant in 9%. The cancer detection rate within theMRI lesionwas 57% (60/104). As the PI-RADS score increased, the detection rate of clinically significant cancer increased (p<0.001, chi squared test). The cancer detection rate for PI-RADS 3 was 31%, PI-RADS 4 was 64% and PI-RADS 5 was 82%. Overall, when MRI-guided biopsy was combined with transperineal sector biopsy the cancer detection rate was 68% (70/104). CONCLUSIONS: MRI-guided prostate biopsies provide good detection rates particularly in PI-RADS 4 and 5 lesions. However, there is ‘MRI Invisible’ cancer which will be missed if biopsies are limited to the lesion only. The highest cancer detection rate was achieved by combining MRI-guided biopsy with transperineal biopsy to assess the rest of the prostate. MRI-targeted biopsy techniques should incorporate a systematic biopsy protocol to identify all clinical significant disease. The presence of ‘MRI Invisible’ cancer also has implications for the planning of focal or targeted therapies.

(329) - The Prevalence of the Frailty Syndrome in Advanced Heart and Lung Failure

s S125 in the increased use of ECMO as a bridge to transplant to minimise waitlist mortality. There is limited data on the use of ECMO prior to LTx in paediatrics and physical outcomes are not well documented. This study aims to evaluate the short-term physical outcomes of pts bridged to LTx via ECMO. Methods: A retrospective analysis was performed of all paediatric pts at a tertiary hospital (n= 4, median age= 13) who underwent bilateral sequential lung transplant (BSLTx) following ECMO support. Indications for transplant included cystic fibrosis (n= 1), acute lung injury (1) and pulmonary vascular disease (2). All pts participated in a supervised, intensive outpatient (OP) exercise rehabilitation program for 1 hour, 3 times a week for 1-3 months. 6 minute walk test (6MWT) results were compared post rehabilitation to a paired cohort of 6 paediatric pts who underwent BSLTx without the need for ECMO pre LTx. Results: Survival at 3 months was 100% for both groups. All pts were female. ECMO support (VA n= 2, VV n= 2) was for a median of 18 days (range 2-35). Median time spent in OP rehabilitation for both groups was 72.5 days (ECMO range 37-80, control 42-77), p= 0.67. 6MWT results on discharge from OP rehabilitation tended to be higher in the ECMO group: median 520m (range 351-557) vs non-ECMO: median 476m (339-503), p= 0.13. All pts progressed to high level physical ability, denoted by jogging or running independently post rehabilitation. Conclusion: The use of ECMO as a bridge to LTx was not shown to be detrimental to short-term physical outcomes based on 6MWT distance. According to available literature, 5/10 pts (3 ECMO, 2 control) achieved a 6MWT distance within the range for healthy paediatrics. Future challenges will include the increasing use of ECMO, in particular, ambulatory circuitry, and the effect of this on short and long-term physical outcomes post paediatric LTx.

(326) - Is MFI Value a Solid Ground to Listing Unacceptable HLA Antigens

s S125 in the increased use of ECMO as a bridge to transplant to minimise waitlist mortality. There is limited data on the use of ECMO prior to LTx in paediatrics and physical outcomes are not well documented. This study aims to evaluate the short-term physical outcomes of pts bridged to LTx via ECMO. Methods: A retrospective analysis was performed of all paediatric pts at a tertiary hospital (n= 4, median age= 13) who underwent bilateral sequential lung transplant (BSLTx) following ECMO support. Indications for transplant included cystic fibrosis (n= 1), acute lung injury (1) and pulmonary vascular disease (2). All pts participated in a supervised, intensive outpatient (OP) exercise rehabilitation program for 1 hour, 3 times a week for 1-3 months. 6 minute walk test (6MWT) results were compared post rehabilitation to a paired cohort of 6 paediatric pts who underwent BSLTx without the need for ECMO pre LTx. Results: Survival at 3 months was 100% for both groups. All pts were female. ECMO support (VA n= 2, VV n= 2) was for a median of 18 days (range 2-35). Median time spent in OP rehabilitation for both groups was 72.5 days (ECMO range 37-80, control 42-77), p= 0.67. 6MWT results on discharge from OP rehabilitation tended to be higher in the ECMO group: median 520m (range 351-557) vs non-ECMO: median 476m (339-503), p= 0.13. All pts progressed to high level physical ability, denoted by jogging or running independently post rehabilitation. Conclusion: The use of ECMO as a bridge to LTx was not shown to be detrimental to short-term physical outcomes based on 6MWT distance. According to available literature, 5/10 pts (3 ECMO, 2 control) achieved a 6MWT distance within the range for healthy paediatrics. Future challenges will include the increasing use of ECMO, in particular, ambulatory circuitry, and the effect of this on short and long-term physical outcomes post paediatric LTx.

EUS‐guided 22‐gauge fine‐needle aspiration versus core biopsy needle in the evaluation of solid pancreatic neoplasms

Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) is widely used for diagnosis of pancreatic lesions. The Echotip Procore Needle (Wilson-Cook Medical) is a new 22G fine biopsy needle (FNB) for obtaining core biopsy material at time of EUS. This study aimed to compare the technical and diagnostic performance of conventional FNA and FNB. Thirty-two patients met the design criteria for this prospective paired cohort study. All lesions sampled were solid (non-cystic) pancreatic masses by EUS appearance. Patients were randomized to receive FNA or FNB by first attempt. A cytopathologist performed on-site evaluations. Samples were assessed for accuracy of diagnosis, cellularity, contamination, and sufficiency for ancillary studies. Technical and diagnostic performances were compared. Compared to FNA, there was a statistically significant decreased ability of FNB to achieve a diagnosis (FNA 93.8%, FNB 28.1%, P < 0.001). FNB was diagnostically superior to FNA in 1 of 32 cases. Technical failures were observed in five cases due to resistance to advancement of the FNB needle. Regarding operator perceived ease-of-use, FNA outperformed FNB (P < 0.001). Eight cases had insufficient FNB material to survive tissue processing. There was no significant difference in mean specimen cellularity between devices. FNA samples showed an increased amount of contaminant (P = 0.036) but were more sufficient for ancillary studies (P = 0.502). Although deemed comparable to FNA when providing material for cytology, the pledged advantage of FNB acting like a core biopsy needle was not apparent in our series. Additional studies are needed before routine adoption of 22G FNB can be recommended.

Baseline Levels and Temporal Stability of 27 Multiplexed Serum Cytokine Concentrations in Healthy Subjects

BackgroundCytokines are humoral molecules that elicit regulatory function in immunologic pathways. The level and type of cytokine production has become critical in distinguishing physiologic from pathologic immune conditions. Cytokine profiling has become an important biomarker discovery tool in monitoring of the immune system. However, the variations in cytokine levels in individual subjects over time in healthy individuals have not been extensively studied. In this study, we use multiplex bead arrays to evaluate 27 analytes in paired serum samples taken seven days apart from 144 healthy individuals in order to assess variations over a short time period.MethodsFluorescent bead-based immunoassay (Luminex) was used to measure 27 analytes in serum samples. Measurements were performed on matched samples from 144 healthy donors. To assess inter-plate variability, one arbitrarily selected serum sample was analyzed on each of the first ten plates as bridge sample. ResultsUsing the bridge sample, we showed minimal inter-plate variations in the measurement of most analytes. In measurement of cytokines from the 144 patients at two time points, we found that three cytokines (IL-2, IL-15 and GM-CSF) were undetectable and five analytes (RANTES, MCP-1, VEGF, MIP-1β and PDGF-BB) showed significant difference in concentrations at Day 0 compared to Day 7. ConclusionsThe current study demonstrated higher variations in cytokine levels among individuals than were observed for samples obtained one week apart from identical donors. These data suggest that a serum sample from each subject for use as a baseline measurement is a better control for clinical trials rather than sera from a paired cohort.

Abnormal vaginal bleeding after epidural steroid injection: a paired observation cohort study

The use of epidural steroid injections has increased dramatically, but knowledge of potential adverse effects is lacking. An association between steroid injection and subsequent abnormal vaginal bleeding has been suspected clinically, but evidence has been limited to anecdotal reports. Paired observational retrospective cohort study using electronic medical records from a large integrated health care system. Participants were all nonhysterectomized women who underwent epidural steroid injections in 2011. For each steroid injection, encounters for abnormal vaginal bleeding during the 60 days preceding and 60 days after the injection were compared as paired observations. For women found to have bleeding, medical records review was performed to examine menopausal status and bleeding evaluation outcomes. Among 8166 epidural steroid injection procedures performed on 6926 nonhysterectomized women, 201 (2.5%) procedures were followed by at least 1 outpatient visit for abnormal vaginal bleeding. Women were 2.8 times more likely to present with abnormal vaginal bleeding during the postinjection period compared with the preinjection period (P < .0001). Of the 197 women with postinjection bleeding, 137 (70%) were premenopausal and 60 (30%) were postmenopausal. Postinjection bleeding prompted endometrial biopsy evaluation in 103 (52%) cases, with benign findings for 100% of premenopausal women (59/59) and 95% of postmenopausal women (42/44). Epidural steroid injections are associated with subsequent abnormal vaginal bleeding for both premenopausal and postmenopausal women. Women undergoing epidural steroid injection should be advised of abnormal bleeding as a potential adverse effect and providers should be aware of this association when evaluating abnormal bleeding.

GATA3 expression in breast carcinoma: utility in triple-negative, sarcomatoid, and metastatic carcinomas

GATA3 plays an integral role in breast luminal cell differentiation and is implicated in breast cancer progression. GATA3 immunohistochemistry is a useful marker of breast cancer; however, its use in specific subtypes is unclear. Here, we evaluate GATA3 expression in 86 invasive ductal carcinomas including triple-negative, Her-2, and luminal subtypes, in addition to 13 metaplastic carcinomas and in 34 fibroepithelial neoplasms. In addition, we report GATA3 expression in matched primary and metastatic breast carcinomas in 30 patients with known estrogen receptor (ER), progesterone receptor (PR), and Her-2 status, including 5 with ER and/or PR loss from primary to metastasis. Tissue microarrays containing 5 to 10 cores per tumor were stained for GATA3, scored as follows: 0 (0-5%), 1+ (6%-25%), 2+ (26%-50%), 3+ (51%-75%), and 4+ (>75%). GATA3 labeling was seen in 67% (66/99) of primary ductal carcinomas including 43% of triple-negative and 54% of metaplastic carcinomas. In contrast, stromal GATA3 labeling was seen in only 1 fibroepithelial neoplasm. GATA3 labeling was seen in 90% (27/30) of primary breast carcinomas in the paired cohort, including 67% of triple-negative carcinomas. GATA3 labeling was overwhelmingly maintained in paired metastases. Notably, GATA3 was maintained in all "luminal loss" metastases, which showed ER and/or PR loss. In conclusion, GATA3 expression is maintained between matched primary and metastatic carcinomas including ER-negative cases. GATA3 can be particularly useful as a marker for metastatic breast carcinoma, especially triple-negative and metaplastic carcinomas, which lack specific markers of mammary origin. Finally, GATA3 labeling may help distinguish metaplastic carcinoma from malignant phyllodes tumors.

444: Outcomes of laser photocoagulation for feto-fetal transfusion syndrome in dichorionic-triamniotic triplet gestations as compared to monochorionic-diamniotic twins

To compare the outcomes of dichorionic-triamniotic (DC,TA) triplet gestations with the outcomes of a paired cohort of monochorionic-diamniotic twin (MC,DA) gestations undergoing laser photocoagulation of placental anastomosis for treatment of feto-fetal transfusion syndrome.

Multiparametric magnetic resonance imaging in the diagnosis and characterisation of prostate cancer: The PROMIS study.

62 Background: Over 1 million US men undergo prostate biopsy each year. About two thirds will test negative. This negative status is uncertain as up to a quarter will have prostate cancer on repeat biopsy. Further, a positive result often fails to discriminate clinically significant from insignificant disease. PROMIS will investigate whether multi-parametric MRI (mpMRI) can discriminate between men with no cancer or clinically insignificant cancer versus those with clinically significant cancer, using template prostate mapping (TPM) biopsies as the most accurate and appropriate reference test in men referred for biopsy. Methods: PROMIS is a multicenter, validating paired cohort study designed to include up to 714 men who will have an mpMRI (index test) and, under general anaesthetic, undergo 10-12 core TRUS biopsy (standard test) and TPM with sampling every 5mm throughout the prostate (reference test). All tests will be assessed independently. Clinically significant cancer on TPM is defined as dominant Gleason pattern ≥4 and/or cancer core length of ≥6mm. PROMIS is powered to measure an absolute increase in sensitivity from 48%, on TRUS biopsy, to at least 70%, by mpMRI. Results: The study will report the proportion of men (i) who could safely avoid biopsy and (ii) correctly identified by mpMRI to have clinically significant prostate cancer. mpMRI will be assessed using an ordinal score of 1 to 5 (highly likely benign - highly likely malignant), with a score ≥3 indicating a positive mpMRI for presence of clinically significant cancer. Secondary objectives will include outcomes based on another definition of clinically significant cancer (non-dominant Gleason pattern ≥4 and/or cancer core length ≥4mm), and cost-effectiveness of using mpMRI prior to biopsy. Conclusions: PROMIS will provide level 1 evidence by comparing mpMRI with the current standard, TRUS biopsy, using an accurate reference test that can be applied to all men advised to have a biopsy. If successful, mpMRI could in the future allow between a quarter and a half of US men referred to safely avoid a biopsy and/or improve the detection of clinically significant cancer by 20%.

Diagnostic accuracy of conventional and cholangioscopy-guided sampling of indeterminate biliary lesions at the time of ERCP: a prospective, long-term follow-up study

The diagnostic accuracy of cholangioscopy-guided sampling has not been rigorously evaluated. To prospectively evaluate the accuracy of cholangioscopy-guided mini-forceps sampling and compare it with standard cytology brushings and forceps biopsies for the tissue diagnosis of indeterminate biliary lesions. Prospective, long-term follow-up, paired design cohort study. Tertiary center. Patients undergoing cholangioscopy for the evaluation of indeterminate biliary lesions. Each patient underwent triple sampling with cholangioscopy-guided mini-forceps, cytology brushing, and standard forceps. Diagnostic accuracy of each sampling method compared with the patient final status (cancer vs no cancer). A total of 26 patients (17 cancer positive/9 cancer negative) were enrolled. The mean follow-up in the patients with no cancer was 21.78 (SD ±6.78) months. The procedure was technically successful in all cases (100%). Sample quality was adequate in 25 of 26 (96.2%) of the cytology brushings, in 26 of 26 (100%) of the standard forceps biopsies, and in 25 of 26 (96.2%) of the mini-forceps biopsies. The sensitivity, accuracy, and negative predictive values were 5.9%, 38.5%, and 36% for standard cytology brushings; 29.4%, 53.8%, and 42.8% for standard forceps biopsies; and 76.5%, 84.6%, and 69.2% for mini-forceps biopsies, respectively. When comparing the 3 methods of sampling, mini-forceps biopsy provided significantly better sensitivity and overall accuracy compared with standard cytology brushing (P < .0001) and standard forceps biopsy (P = .0215). Potential for selection bias. Cholangioscopy-guided biopsies of indeterminate biliary lesions have significantly higher accuracy compared with ERCP-guided cytology brushings and standard forceps biopsies, but negative findings on mini-forceps biopsy cannot rule out malignancy with a high degree of certainty. ( NCT01227382.).