Two-sided Wilcoxon Rank Sum Test Research Articles

Ultra-Sensitive Detection of Circulating Serum microRNAs (miRNAs) in Diffuse Large B-Cell Lymphoma (DLBCL) Patient-Derived Xenograft (PDX) Models and Correlation with Disease Status in DLBCL Patient

Background: MiRNAs are small non-coding RNAs that regulate post-transcriptional gene expression, contribute to various facets of cancer pathogenesis, and may serve as a sensitive diagnostic platform as well as potential novel therapeutic targets. We recently identified a 9 miRNA (miR-15a, let-7c, let-7b, miR-27a, miR10b, miR-18a, miR130a, miR24, and miR155) signature in serum of Smurf2T/T mice that was detectable many months prior to the formation of visible DLBCL (Beheshti A et al PLOS One, 2017). We hypothesized that this circulating miRNA signature would also correlate with DLBCL status in PDX models and ultimately patients (pts).Methods: We performed droplet digital PCR (ddPCR), which allows for rapid quantification of single miRNA molecules. First, we examined the 9 aforementioned miRNAs in serum collected from mice xenografted with 5 DLBCL PDXs (Townsend et al. Cancer Cell 2016) and age-matched NSG mice without xenografts as controls. We also quantified the amount of miRNAs directly from all PDX cell lines utilized and 2 commercially available DLBCL cell lines for comparison. Next, we collected and analyzed serum from 86 pts with DLBCL in the following conditions: pre-treatment with first-line therapy (n=11); progression after treatment (n=7); during treatment (n=16); and in complete remission (n=52). Seventeen healthy non-tumor bearing individuals were used as controls. Median age of healthy controls was 52 years (range, 29-70) vs 64 years (range, 21-87) for DLBCL pts (p<0.001; Wilcoxon rank-sum test). 69% of DLBCL pts were male vs 35% of healthy controls (p=0.012; Fisher exact test).Results: The miRNA signature was enriched in DLBCL PDX mice with high expression confirmed in the 9 miRNAs, including MYC single and double hit models. The overall mean amounts of the miRNAs present in the serum of PDX bearing mice were mostly equivalent to the amount present in the original PDX cell line (Fig 1), with exception of one PDX cell line (DFBL-75549) that consistently had 100x more miRNA present in the serum of the mice vs associated cell line. In addition, we identified significantly increased circulating levels of the same miRNA signature in the serum of DLBCL pts (Table 1) (two-sided Wilcoxon rank-sum test, p<0.05) with similar patterns of change as seen in the murine models. Interestingly, higher circulating levels of let-7b were associated with a higher stage at diagnosis (stage I-II vs III-IV, median 54.0 vs. 163.2; P=0.007) and higher levels of both miR-27a and miR-24 were associated with having a MYC rearrangement (median 20.0 vs. 4.8; P=0.003; median 58.4 vs. 24.4; P=0.046). Higher levels of miR-18a were associated with Myc positivity by immunohistochemistry (0-9% of cells vs. 10-59% vs. ≥60%, median 3.3 vs. 2.8 vs. 8.8; P=0.030) as well as having received a prior therapy for DLBCL (median 4.8 for previously treated vs. 1.1 for untreated; P=0.016). Using the 23 on-treatment and progression samples compared with healthy samples, we selected cut-points based on the Youden Index from receiver operating curve (ROC) analysis and were able to classify the remission samples with an accuracy of 46%-88%. miR-24 performed the best in classification with a sensitivity of 85% and a specificity of 100%, while 6 of the 9 miRs had a classification rate >80%. Using a 5 miR signature with cut-points selected from recursive partitioning, we were able to classify remission samples with an accuracy of 91% (sensitivity 90%, specificity 94%).Conclusions: Altogether, ultrasensitive detection of circulating miRNAs originally identified in a lymphoma xenograft knockout model was readily detectable and highly elevated in DLBCL PDX models. Additionally, there were significantly increased circulating levels of the miRNA signature from the serum of DLBCL pts. Particular miRs were associated with pt stage and the presence of MYC overexpression or rearrangement in pts with DLBCL. Furthermore, circulating miRNAs were able to reliably distinguish DLBCL pts in remission from healthy controls based on a novel 5 miR signature. Validation in additional cohorts is needed to confirm whether miRNA quantification from serum may be a broadly applicable strategy for diagnosis, classification and response assessment among pts with DLBCL. [Display omitted] DisclosuresWeinstock:Astra Zeneca, JAX, Samumed, Regeneron, Sun Pharma, Prescient: Patents & Royalties; Genentech/Roche, Monsanto: Consultancy; Novartis: Consultancy, Research Funding; Novartis, Astra Zeneca, Abbvie, Aileron, Surface Oncology, Daiichi Sankyo: Research Funding; Novartis, Dragonfly, Travera, DxTerity, Travera: Consultancy; Travera: Equity Ownership. Evens:Pharmacyclics International DMC: Membership on an entity's Board of Directors or advisory committees; Bayer: Consultancy; Seattle Genetics, Inc.: Membership on an entity's Board of Directors or advisory committees, Research Funding; Affimed: Consultancy; Abbvie: Consultancy; Tesaro: Research Funding; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees; Janssen: Consultancy; Novartis: Consultancy; Acerta: Consultancy.

Optoacoustic Breast Imaging: Imaging-Pathology Correlation of Optoacoustic Features in Benign and Malignant Breast Masses

Optoacoustic ultrasound breast imaging is a fused anatomic and functional modality that shows morphologic features, as well as hemoglobin amount and relative oxygenation within and around breast masses. The purpose of this study is to investigate the positive predictive value (PPV) of optoacoustic ultrasound features in benign and malignant masses. In this study, 92 masses assessed as BI-RADS category 3, 4, or 5 in 94 subjects were imaged with optoacoustic ultrasound. Each mass was scored by seven blinded independent readers according to three internal features in the tumor interior and two external features in its boundary zone and periphery. Mean and median optoacoustic ultrasound scores were compared with histologic findings for biopsied masses and nonbiopsied BI-RADS category 3 masses, which were considered benign if they were stable at 12-month follow-up. Statistical significance was analyzed using a two-sided Wilcoxon rank sum test with a 0.05 significance level. Mean and median optoacoustic ultrasound scores for all individual internal and external features, as well as summed scores, were higher for malignant masses than for benign masses (p < 0.0001). High external scores, indicating increased hemoglobin and deoxygenation and abnormal vessel morphologic features in the tumor boundary zone and periphery, better distinguished benign from malignant masses than did high internal scores reflecting increased hemoglobin and deoxygenation within the tumor interior. High optoacoustic ultrasound scores, particularly those based on external features in the boundary zone and periphery of breast masses, have high PPVs for malignancy and, conversely, low optoacoustic ultrasound scores have low PPV for malignancy. The functional component of optoacoustic ultrasound may help to overcome some of the limitations of morphologic overlap in the distinction of benign and malignant masses.

Rehydration before wet fixation in conventional body fluid cytology - An 18-year experience.

In conventional cytology, preparation of a specimen by wet fixation for Papanicolaou stain is potentially subject to dry effect or cell loss which may make cytologic interpretation difficult or even impossible. We have been routinely making an additional smear for rehydration with normal saline (rehydration method) before wet fixation to overcome the above shortcomings. We reviewed malignant pleural effusion and ascites 15 cases each in our cytology laboratory over the past 1year. Four slides of each specimen were made. Two were air-dried for Liu's stain (a Romanowsky stain) and the other two were wet-fixed for Papanicolaou stain. The air-dried smears were also served as retained cellularity control. One of the two wet-fixed smears was processed as a control of preservation of nuclear detail whereas the other one stayed air-dried for 10minutes and then covered with normal saline (rehydration method) for 80seconds before wet fixation. There was minor cell loss (P=.032). The cells appeared larger with good preservation of nuclear detail (P<.0001 by two-sided Wilcoxon rank sum test) but no red blood cells retained on the slide after rehydration. The rehydration method can effectively minimise cell loss, enlarge and preserve the cytological features of malignant cells with haemolysis. This method is simple, practical and good for cytological screening for tumour cells and interpretation especially in a bloody smear. We recommend that the rehydration method be part of traditional cytopreparatory work of wet fixation for Papanicolaou stain in conventional body fluid cytology.

Effectiveness of Transmastoid Plugging for Semicircular Canal Dehiscence Syndrome.

Objectives (1) Evaluate changes in subjective symptoms in patients following transmastoid canal plugging for superior semicircular canal dehiscence (SSCD) syndrome. (2) Quantify changes in hearing in patients who have undergone transmastoid canal plugging for SSCD syndrome. Study Design Case series with chart review. Setting Single tertiary care institution. Subjects and Methods We retrospectively reviewed patients with SSCD who underwent repair with canal plugging via a transmastoid approach between January 2012 and January 2017. Symptom severity was assessed prospectively (autophony, sound/pressure-induced vertigo, disequilibrium, aural fullness, and pulsatile tinnitus) and after surgery. Pure-tone and speech audiometry were measured before and after surgery. Two-sided Wilcoxon rank-sum tests were used to evaluate changes in subjective symptoms and audiometric outcomes. Results Seventeen patients (19 ears) met inclusion criteria. The superior canal was successfully plugged via the transmastoid approach in all cases. Patients reported a statistically significant improvement in autophony, vertigo, aural fullness, and pulsatile tinnitus ( P < .01), without significant improvement in disequilibrium rating ( P = .06). There were no changes noted in pure-tone average or word recognition score; however, there was a statistically significant improvement in air-bone gap at 250 Hz of 10.9 dB ( P = .04) with 12.9-dB improvement in air conduction thresholds ( P = .02) and no difference (0.9 dB, P = .9) in bone conduction thresholds. Conclusion In our study, patients with SSCD demonstrated excellent hearing outcomes and resolution of most otologic symptoms after surgical repair. Transmastoid canal plugging, which has been described to date only in smaller case series, is a safe and effective alternative to the traditional middle cranial fossa approach.

Analyzing Volatile Anesthetic Consumption by Auditing Fresh Gas Flow: An Observational Study at an Academic Hospital.

BackgroundIn a climate of cost containment, it is critical to analyze and optimize all perioperative variable costs. Fresh gas flow is one important variable that determines utilization of inhalational agents and can be tightly controlled by the anesthesia provider. Manufacturers of inhalational agents have recommendations for minimum gas flow for their respective agents. Any gas flow above these recommendations is considered misuse and leads to unnecessary expense. The purpose of this study was to characterize and quantify the excess use of inhalational agents by analyzing fresh gas flow rates for long duration cases.MethodsOver a span of three months, operating room records were analyzed for all procedures lasting greater than 4 hours. End tidal inhalation agent percentage for Sevoflurane and Isoflurane and fresh gas flows were analyzed. 303 unique patients with at least 4 hours of anesthesia time were included. Analysis excluded the first and last 30 minutes of all anesthetics to account for need for higher gas flows during induction/emergence of anesthesia. 152 patients received sevoflurane alone. 33 patients received isoflurane alone. 107 patients received both isoflurane and sevoflurane and were included in sevoflurane group given the higher gas flow needs of sevoflurane. 11 patients received neither agent and were excluded from analysis. We proceed with n = 292 unique patients. (259 in Sevo, 33 in iso) We used the two-sided one sample t-test setting 2 ml/min as the null for sevo and 1 ml/min as the null for iso; we ran analysis using a nonparametric test that didn’t require the fresh gas flow to be normally distributed - the two-sided one-sample Wilcoxon rank-sum test: p value = < 0.0001.ResultsThe results of our study revealed a sevoflurane (n = 259) mean fresh gas flow (L/min) 2.55 (95% CI, 2.45-2.66) - significantly different from null of 2 ml/min (p < 0.0001). Isoflurane (n = 33) mean fresh gas flows (L/min) 2.33 (95% CI, 2.00-2.66) - significantly different from null of 1 l/min (p < 0.0001).ConclusionManufacturer recommendation for sevoflurane is to maintain gas flows 1-2 l/min and Isoflurane at above 1 l/min. Given these recommendations, the anesthesia providers delivered fresh gas flows at least 28% higher than necessary for sevoflurane and at least 130% greater than necessary for isoflurane anesthetics that lasted greater than 4 hours. This is an area where cost reduction can be readily achieved. Future plans to realize a reduction in inhalational agent utilization include education of the benefits of fresh gas flow and instituting a low fresh gas flow policy.

Abstract PR463

Background & Objectives: Background: In a climate of cost containment, it is critical to analyze and optimize all perioperative variable costs. Fresh gas flow is one important variable that determines utilization of inhalational agents and can be tightly controlled by the anesthesia provider. Manufacturers of inhalational agents have recommendations for minimum gas flow for their respective agents. Any gas flow above these recommendations is considered misuse and leads to unnecessary expense. The purpose of this study was to characterize and quantify the excess use of inhalational agents by analyzing fresh gas flow rates for long duration cases. Materials & Methods: Methods: Over a span of three months, operating room records were analyzed for all procedures lasting greater than 4 hours. End tidal inhalation agent percentage for Sevoflurane and Isoflurane and fresh gas flows were analyzed. 303 unique patients with at least 4 hours of anesthesia time were included. Analysis excluded the first and last 30 minutes of all anesthetics to account for need for higher gas flows during induction/emergence of anesthesia. 152 patients received sevoflurane alone. 33 patients received isoflurane alone. 107 patients received both isoflurane and sevoflurane and were included in sevoflurane group given the higher gas flow needs of sevoflurane. 11 patients received neither agent and were excluded from analysis. We proceed with n=292 unique patients. (259 in Sevo, 33 in iso) We used the two-sided one sample t-test setting 2ml/min as the null for sevo and 1ml/min as the null for iso; we ran analysis using a nonparametric test that didn’t require the fresh gas flow to be normally distributed-- the two-sided one-sample Wilcoxon rank-sum test: p=value<0.0001 Results: Results: The results of our study revealed a sevoflurane (n=259) mean fresh gas flow (l/min) 2.55 (95%CI, 2.45-2.66)--significantly different from null of 2ml/min:p<0.0001. Isoflurane (n=33) mean fresh gas flows (l/min) 2.33 (95%CI, 2.00-2.66)--significantly different from null of 1l/min:p<0.0001. Conclusion: Conclusion: Manufacturer recommendation for sevoflurane is to maintain gas flows 1-2 l/min and Isoflurane at above 1l/min. Given these recommendations, the anesthesia providers delivered fresh gas flows at least 28% higher than necessary for sevoflurane and at least 130% greater than necessary for isoflurane anesthetics that lasted greater than 4 hours. This is an area where cost reduction can be readily achieved. Future plans to realize a reduction in inhalational agent utilization include education of the benefits of fresh gas flow and instituting a low fresh gas flow policy. Disclosure of Interest: None declared

MR Imaging-derived Regional Pulmonary Parenchymal Perfusion and Cardiac Function for Monitoring Patients with Chronic Thromboembolic Pulmonary Hypertension before and after Pulmonary Endarterectomy.

Purpose To evaluate surgical success after pulmonary endarterectomy (PEA) by means of cardiopulmonary magnetic resonance (MR) imaging. Materials and Methods In this institutional review board-approved study, 20 patients with chronic thromboembolic pulmonary hypertension were examined at 1.5 T with a dynamic contrast material-enhanced three-dimensional fast low-angle shot sequence before and 12 days after PEA (25th-75th percentile range, 11-16 days). Lung segments were evaluated visually before PEA for parenchymal hypoperfused segments. Pulmonary blood flow (PBF), first-pass bolus kinetic parameters, and biventricular mass and function were determined. Mean pulmonary artery pressure (mPAP) and 6-minute walking distance were measured before and after PEA. The Shapiro-Wilk test, paired two-sided Wilcoxon rank sum test, Spearman ρ correlation, and multiple linear regression analysis were performed. Results Two weeks after PEA, regional PBF increased 66% in the total lung from 32.7 to 54.2 mL/min/100 mL (P = .0002). However, after adjustment for cardiac output, this change was not evident anymore (increase of 7% from 7.03 to 7.54 mL/min/100 mL/L/min, P = .1). Only in the lower lobes, a significant increase in PBF after cardiac output adjustment remained: a 16% increase in the right lower lobe from 7.53 to 8.71 mL/min/100 mL (P = .01) and a 14% increase in the left lower lobe from 7.42 to 8.47 mL/min/100 mL/L/min (P < .05). Right ventricular mass and function also improved. mPAP decreased from 46 to 24 mm Hg (P < .0001). Six-minute walking distance increased from 390 to 467 m (P = .02) 5 months after PEA. Percentage change of mPAP and PBF in the lower lobe tended to be significant predictors of percentage change in 6-minute walking distance (β = -1.79 [P = .054] and β = 0.45 [P = .076], respectively) in multiple linear regression analysis. Conclusion Improvement of PBF after PEA was observed predominantly in the lower lungs, and the magnitude of improvement of PBF in the lower lobes correlated with the improvement in exercise capacity, reflecting surgical success. (©) RSNA, 2016.

Status of the Parkinson's disease gene family expression in non-small-cell lung cancer.

BackgroundThe purpose of this study is to detect the Parkinson’s disease gene family mRNA relative expression in the non-small-cell lung cancer (NSCLC) tumor tissue and analyze the association between tumor characteristics and the Parkinson’s disease gene family.MethodsTumor tissue and tumor-adjacent tissue of 114 NSCLC patients were collected and SYBR quantitative analysis was used to detect the relative expression level of nine Parkinson’s disease gene mRNAs. Then, paired sample test, two-sided Student’s t-test, or two-sided Wilcoxon rank sum test was performed to analyze the mRNA relative expression level of nine Parkinson’s disease gene mRNAs in different gender, tumor histology, and tumor stage.ResultsOverexpression in the tumors was detected in 46/114 (40.35 %) PARK1/4, 74/114 (64.91 %) PARK2, 104/114 (91.23 %) PARK5, 95/114 (83.33 %) PARK6, 80/114 (70.18 %) PARK7, 55/114 (48.25 %) PARK8, 100/114 (87.72 %) PARK9, 55/114 (48.25 %) PARK15, and 99/114 (86.84 %) glucocerebrosidase (GBA). Five genes PARK5 (91.23 %), PARK6 (83.33 %), PARK7 (70.18 %), PARK9 (87.72 %), and GBA (86.84 %) were supposed to be overexpressed in the lung tumor tissues compared with tumor-adjacent tissues. There was no significant difference in PARK1/4, PARK2, PARK5, PARK9, and GBA mRNA expression by different tumor stage, whereas, PARK6, PARK7, PARK8, and PARK15 mRNA expression were found to have significant difference in the comparison of different tumor stages. The expression of PARK6 (P = 0.01, P = 0.03) and PARK15 (P < 0.001, P < 0.001) were significantly higher in stages I and II when compared with stage III, respectively. NSCLC patients in stage I showed the higher expression PARK7 compared to the patients in stage II (P = 0.003).ConclusionsThe high expression of PARK6, PARK7, and PARK15 might lead to the occurrence of a primary NSCLC tumor, and the tumor with a decreasing expression of these three genes tends to be stages II and III. The results of our study indicate that the Parkinson’s disease gene family may be a potential marker for the prediction of NSCLC.Electronic supplementary materialThe online version of this article (doi:10.1186/s12957-015-0646-y) contains supplementary material, which is available to authorized users.

The effects of soy supplementation on gene expression in breast cancer: a randomized placebo-controlled study.

There are conflicting reports on the impact of soy on breast carcinogenesis. This study examines the effects of soy supplementation on breast cancer-related genes and pathways. Women (n = 140) with early-stage breast cancer were randomly assigned to soy protein supplementation (n = 70) or placebo (n = 70) for 7 to 30 days, from diagnosis until surgery. Adherence was determined by plasma isoflavones: genistein and daidzein. Gene expression changes were evaluated by NanoString in pre- and posttreatment tumor tissue. Genome-wide expression analysis was performed on posttreatment tissue. Proliferation (Ki67) and apoptosis (Cas3) were assessed by immunohistochemistry. Plasma isoflavones rose in the soy group (two-sided Wilcoxon rank-sum test, P < .001) and did not change in the placebo group. In paired analysis of pre- and posttreatment samples, 21 genes (out of 202) showed altered expression (two-sided Student's t-test, P < .05). Several genes including FANCC and UGT2A1 revealed different magnitude and direction of expression changes between the two groups (two-sided Student's t-test, P < .05). A high-genistein signature consisting of 126 differentially expressed genes was identified from microarray analysis of tumors. This signature was characterized by overexpression (>2-fold) of cell cycle transcripts, including those that promote cell proliferation, such as FGFR2, E2F5, BUB1, CCNB2, MYBL2, CDK1, and CDC20 (P < .01). Soy intake did not result in statistically significant changes in Ki67 or Cas3. Gene expression associated with soy intake and high plasma genistein defines a signature characterized by overexpression of FGFR2 and genes that drive cell cycle and proliferation pathways. These findings raise the concerns that in a subset of women soy could adversely affect gene expression in breast cancer.

Global field synchrony during general anaesthesia

BackgroundAnaesthetic agents may disrupt consciousness by inhibiting long-range synchronization of brain activity. In the current study, the patterns of widespread and spatially localized synchrony during anaesthesia are investigated using a measure called global field synchrony (GFS). MethodsThe EEG obtained during routine surgery in 29 patients was analysed with GFS over the following frequency bands: δ (1.5–3.5 Hz), θ (3.5–7.5 Hz), α1 (8–10 Hz), α2 (10.5–12 Hz), β1 (12.5–18 Hz), β2 (18.5–21 Hz), β3 (21.5–30 Hz), γ1 (30.5–40 Hz), and γ2 (60–80 Hz). In addition, localized GFS estimations over aggregate brain areas were performed. GFS was estimated over 2 s non-overlapping windows. The differences in GFS values between ‘wakefulness’ and ‘anaesthesia’ were assessed with the two-sided Wilcoxon rank-sum tests (α=0.05). ResultsAnaesthetic administration caused significant GFS changes in all frequency ranges and electrode combinations studied: (i) widespread synchrony increased in the α2 and β1 ranges and decreased in all other ranges, with the exception of α1 and β2, where no specific pattern was identified; and (ii) localized synchrony decreased in all areas in the δ and γ2 ranges, while location-specific changes were observed in the remaining frequency ranges. The most consistent findings were statistically significant decreases over all areas in the γ2 range, with GFS decrease over the central–right temporal being the most consistent change. ConclusionsSignificant frequency- and location-dependent changes in GFS were induced by anaesthetic administration, with more robust changes identified in the γ range. GFS can act as an aid for further and more detailed analysis regarding the particular combinations of frequency ranges and spatial locations that are most informative for the study of anaesthetic-induced unconsciousness.

System-wide Analysis Reveals Intrinsically Disordered Proteins Are Prone to Ubiquitylation after Misfolding Stress

Damaged and misfolded proteins that are no longer functional in the cell need to be eliminated. Failure to do so might lead to their accumulation and aggregation, a hallmark of many neurodegenerative diseases. Protein quality control pathways play a major role in the degradation of these proteins, which is mediated mainly by the ubiquitin proteasome system. Despite significant focus on identifying ubiquitin ligases involved in these pathways, along with their substrates, a systems-level understanding of these pathways has been lacking. For instance, as misfolded proteins are rapidly ubiquitylated, unconjugated ubiquitin is rapidly depleted from the cell upon misfolding stress; yet it is unknown whether certain targets compete more efficiently to be ubiquitylated. Using a system-wide approach, we applied statistical and computational methods to identify characteristics enriched among proteins that are further ubiquitylated after heat shock. We discovered that distinct populations of structured and, surprisingly, intrinsically disordered proteins are prone to ubiquitylation. Proteomic analysis revealed that abundant and highly structured proteins constitute the bulk of proteins in the low-solubility fraction after heat shock, but only a portion is ubiquitylated. In contrast, ubiquitylated, intrinsically disordered proteins are enriched in the low-solubility fraction after heat shock. These proteins have a very low abundance in the cell, are rarely encoded by essential genes, and are enriched in binding motifs. In additional experiments, we confirmed that several of the identified intrinsically disordered proteins were ubiquitylated after heat shock and demonstrated for two of them that their disordered regions are important for ubiquitylation after heat shock. We propose that intrinsically disordered regions may be recognized by the protein quality control machinery and thereby facilitate the ubiquitylation of proteins after heat shock.

Swallowing Outcomes for T1‐T4 Oropharyngeal Cancers Treated with TORS: Preliminary Experience

Objectives: 1) Assess swallowing function longitudinally using standardized questionnaires in a cohort of oropharyngeal cancer patients treated with transoral robotic surgery (TORS). 2) Compare the effect of tumor T-stage on swallowing outcomes. Methods: A prospective, institutional review board (IRB)-approved database of all patients undergoing TORS for oropharyngeal cancer at a tertiary-care academic institution was queried from November 2010-October 2012. Demographic data as well as longitudinal assessment of swallowing outcomes were collected. Standardized questionnaires included: University of Washington Quality of Life question 5 (UW-QOLq5), MD Anderson Dysphagia Inventory (MDADI), and Functional Oral Intake Scale (FOIS). Two-sided Wilcoxon signed-rank tests were used to compare baseline values with corresponding 1, 3, 6, and 12 months post-surgery. Two-sided Wilcoxon rank-sum tests were used to test for differences between T-stages. Results: Thirty-nine patients were studied with a mean age of 60.7 (42-92). T-stages included: T1 n = 14, T2 n = 15, T3 n = 6, T4 n = 4. No significant differences were detected at any time between patients with T1-T2 and T3-T4 stage disease on UW-QOLq5, MDADI, or FOIS. Compared to baseline, UW-QOLq5 measures were significantly lower at 1 month ( P = 0.0156), but not at 3 or 6 months. MDADI overall scores were significantly lower than baseline at 1 month ( P = 0.0134), but not at 3, 6, or 12 months. Compared to baseline, FOIS measures were significantly lower at 1 month ( P &lt;.0001), and remained significantly lower at 3 ( P &lt;.0001), 6 ( P &lt;.0001), and 12 months ( P = 0.0039). Conclusions: Results suggests that while swallowing status declines immediately after TORS, it can be expected to approach baseline within 3 months post-surgery.

Absence of anti‐hepatitis B virus (HBV) core in HIV/HBV coinfection with advanced immunosuppression

Hepatitis B virus (HBV) coinfection is frequent among HIV-infected patients and is associated with poorer outcomes related to monoinfection with either virus [1]. These patients may present atypical HBV serological patterns, such as nonreactive anti-HBV core (HBc) with reactive HBV surface antigen (HBsAg) [2]. We investigated the prevalence of HBsAg with nonreactive anti-HBc in a cohort of HBV/HIV-coinfected patients from an AIDS Clinic in Sao Paulo, Brazil. The study population was selected from a cohort of 2412 HIV-positive patients, of whom 120 were HBsAg reactive. Cases were chronically HBV-infected patients with nonreactive anti-HBc, either transiently or persistently. Controls were obtained from the same population, but had persistently reactive anti-HBc. Cases comprised 11 men and one woman (39 - 61 years old), while controls comprised 24 anti-HBcand HBsAg-reactive patients matched by age and gender in a control:case ratio of 2:1. HIV results were obtained using two commercial enzyme immunosorbent assays (Organon Technika, Tournault, Belgium and Embrabio, Sao Paulo, Brazil) and confirmed using GS-HIV-1 Western Blot (Bio-Rad, Hercules, CA). HIV RNA was quantified using the Versant HIV-1 RNA 3.0 bDNA assay (Siemens Healthcare, Erlangen, Germany). CD4 T-lymphocyte counts were determined using the BD Multitest/Trucount (BD Biosciences, San Jose, CA). A microparticle enzyme immunoassay (AxSYM; Abbott Laboratories, North Chicago, IL) was used to detect HBV serological markers. HBV DNA was quantified using real-time polymerase chain reaction [3] and pre-core/core and surface gene sequences were obtained for mutation detection and subgenotyping in HBV DNA-positive cases [4]. For each patient, the mean CD4 count and viral load measurement for all measurements performed 12 months prior to each HBV serological test were calculated. Opportunistic diseases and complications of chronic HBV infection were also reported. The use of HBV-active drugs (lamivudine, tenofovir, entecavir or interferon) prior to the first serological test included in the study for cases and at any time for controls was evaluated. Variables were compared using the two-sided Wilcoxon rank-sum test (for continuous variables) or the two-sided Fisher’s exact test (for categorical variables) with STATA 10.1 (Stata Corporation, College Station, TX), and a significance level of 0.05. We identified 120 (5.0%) chronically HBV-infected patients, 12 (10.0%) of whom presented with nonreactive anti-HBc. A significantly higher proportion of cases were persistently positive for HBeAg, and cases had a lower mean CD4 count compared with controls (Table 1). Amplification of HBV DNA was achieved in three out of seven cases who were available for collection of a new sample, with a mean viral load of 5.67 log HBV DNA copies/mL. HBV subgenotype A1 was detected in all three cases. No mutations in pre-core/core and S gene sequences were identified.

Sex Difference in Normal Thymic Appearance in Adults 20–30 Years of Age

To determine whether there is a sex difference in the appearance of the normal thymus in 20-30-year-old men and women. This retrospective study was approved by the institutional review board and was compliant with HIPAA. The requirement for informed consent was waived. Images and medical records of 238 consecutive subjects without known thymic disease (175 men, 63 women) who underwent chest computed tomography with intravenous contrast material in 2008 were reviewed. Average thymic region of interest (ROI), subjective assessment of thymic attenuation by using a scale of grades 0-3, thymic anteroposterior measurement, and mean maximal thymic lobe thickness were recorded by two independent thoracic radiologists, blinded to subject age and sex. Thymic morphologic characteristics were assessed in consensus. The two-sided Wilcoxon rank-sum test, Student t test, test for linear regression, analysis of covariance, two-way factorial analysis of variance, and continuity-adjusted χ(2) test were used for statistical analysis. There was a significant sex difference in thymic attenuation as measured objectively by using mean thymic ROI measurement (P < .0001) and subjectively by using a scale of grades 0-3 (P < .0001), which held true when corrected for age (P < .0001). A sex difference was also found in regard to the rate of decrease in mean thymic attenuation with age, with men's thymuses exhibiting a significant decrease in attenuation during the decade, unlike women (P = .0479). There was no significant sex difference in mean maximal thymic lobe thickness (P = .8697). A quadrilateral, as opposed to triangular, configuration of the thymus was more common in women than men (P = .0034). There is a significant sex difference in normal thymic appearance in 20-30-year-old men and women. The thymus of 20-30-year-old women typically exhibits higher attenuation and more commonly exhibits a quadrilateral configuration.

Impact of periictal interventions on respiratory dysfunction, postictal EEG suppression, and postictal immobility

Sudden unexpected death in epilepsy (SUDEP) is the leading cause of epilepsy-related mortality. Seizure-related respiratory dysfunction (RD), the duration of postictal generalized electroencephalography (EEG) suppression (PGES), and duration of postictal immobility (PI) may be important in the pathophysiology of SUDEP. Periictal interventions may reduce the risk of SUDEP. We assessed the impact of periictal nursing interventions on RD, PGES, and PI duration in patients with localization-related epilepsy and secondarily generalized convulsions (GCs) recorded during video-EEG telemetry in the epilepsy monitoring unit. Video-EEG data were retrospectively reviewed. Interventions including administration of supplemental oxygen, oropharyngeal suction, and patient repositioning were evaluated. Interventions were performed based on nursing clinical judgment at the bedside and were not randomized. The two-sided Wilcoxon rank-sum test was used to compare GCs with and those without intervention. Robust simple linear regression was used to assess the association between timing of intervention and duration of hypoxemia (SaO(2) < 90%), PGES, and PI using data from only the first GC for each patient. Data from 39 patients with 105 GCs were analyzed. PGES >2 s occurred following 31 GCs in 16 patients. There were 21 GCs with no intervention (NOINT) and 84 GC with interventions (INT). In the INT group, the duration of hypoxemia was shorter (p = 0.0014) when intervention occurred before hypoxemia onset (mean duration 53.1 s) than when intervention was delayed (mean duration 132.42 s). Linear regression indicated that in GCs with nursing interventions, earlier intervention was associated with shorter duration of hypoxemia (p < 0.0001) and shorter duration of PGES (p = 0.0012). Seizure duration (p < 0.0001) and convulsion duration (p = 0.0457) were shorter with earlier intervention. PI duration was longer for GCs with PGES than GCs without PGES (p < 0.0001). The mean delay to first active nonrespiratory movement following GCs with PGES was 251.96 s and for GC without PGES was 66.06 s. The duration of PI was positively associated with lower SaO(2) nadir (p = 0.003) and longer duration of oxygen desaturation (p = 0.0026). There was no association between PI duration and seizure duration (p = 0.773), between PI duration and PGES duration (p = 0.758), or between PI duration and the timing of first intervention relative to seizure onset (p = 0.823). PGES did not occur in the NOINT group. The mean duration of desaturation was longer (110.9 vs. 49.9 s) (p < 0.0001), mean SaO(2) nadir was lower (72.8% vs. 79.7%) (p = 0.0086), and mean end-tidal CO(2) was higher (58.6 vs. 50.3 mmHg) (p = 0.0359) in the INT group compared with the NOINT group. The duration of the seizure or of the convulsive component was not significantly different between the INT and NOINT groups. Early periictal nursing intervention was associated with reduced duration of RD and reduced duration of PGES. These findings suggest the possibility that such interventions may be effective in reducing the risk of SUDEP in the outpatient setting. Validation of these preliminary data with a prospective study is needed before definitive conclusions can be reached regarding the efficacy of periictal interventions in reducing the risk of SUDEP.

Excessive Weight Loss after Sleeve Gastrectomy: A Systematic Review

The clinical significance of sleeve gastrectomy (SG) as a primary bariatric intervention is still under debate. This article aims to systematically analyze excessive weight loss (EWL) in patients after SG. A systematic literature search on SG from the period January 2003 to December 2010 was performed. Data described from systematic reviews dealing with gastric bypass procedures was used as comparator. The final study included 123 papers describing 12,129 patients. Most of the papers describe EWL at 12 months (43.9% of all papers). For SG, the maximum EWL occurred 24 and 36 months postoperatively with a mean EWL of 64.3% (minimum 46.1%, maximum 75.0%) and 66.0% (minimum 60.0%, maximum 77.5%), respectively. At 12 months, the mean EWL in patients receiving SG was significantly lower when compared to patients who underwent gastric bypass (SG 56.1%, gastric bypass 68.3%; p < 0.01, two-sided Wilcoxon test). Although patients with gastric bypass still had higher EWL rates at 24 months compared to patients after SG, these differences were not significant (SG 61.3%, gastric bypass 69.6%; p = 0.09, two-sided Wilcoxon rank-sum test). Reoperations after SG are necessary in 6.8% (range 0.7-25%) of cases with patients receiving SG as a stand alone procedure and in 9.6-28.5% of cases with patients undergoing SG as a planned first stage procedure. SG is an effective bariatric procedure with a lasting effect on EWL. Compared with gastric bypasses, there is no difference in EWL at the time point of 24 months.

PD10-07: microRNA Profiles of Breast Tumors Identifies the miR 17–92 Cluster as a Group of Potentially Essential Oncomirs in Triple-Negative Breast Cancer.

Abstract Introduction: MicroRNAs (miRNAs) are a class of short non-coding RNAs of about 20–24 nucleotides that act post-transcriptionally to modulate gene expression by sequence-specific base-pairing with target mRNAs. While the function of human miRNA remains largely unknown, certain miRNAs have previously been implicated in cancer development and progression. In this study, we investigate the expression of 734 miRNAs across a cohort of 7 normal breast samples and 104 breast cancer tumors including 70 ER−/PR−/HER2−(triple negative, TN), 15 ER+/PR+, and 19 HER2+. Methods: We used the novel Nanostring technology to quantify the expression of 734 miRNAs. Unsupervised hierarchical clustering was performed using miRNAs that were detected as expressed in at least one sample. A two-sided Wilcoxon rank sum test was used to identify miRNAs with a significantly differential expression between two sets of samples. Results: In agreement with previously published results, we found that tumor samples clustered into the luminal and TN subtypes using unsupervised hierarchical clustering on the expressed miRNAs. Interestingly, microRNA expression was similar between HER2 amplified ER+ and HER2 amplified ER−, suggesting a common luminal origin. When we specifically compared luminal (ER+ or HER2+) to TNBC, we found that 50 miRNAs showed significant differential expression. In particular, the miR-17-92 cluster(1) was expressed at significantly higher levels in TN tumors. Within the luminal tumors the miR-17-92 cluster was strongly correlated with proliferation and grade; although miR17-92 was expressed at high level, there was no correlation with proliferation in the TN tumors. miR17-92, which inhibits translation of E2F1(1), is in a regulatory network with myc and E2F to control the balance between proliferation and apoptosis(2). The absence of correlation between miR17-92 and proliferation in the TN tumors suggests disruption of this regulatory network. Indeed, we found a positive correlation between expression of miR17-92 and p16 and inverse correlation with RB across the tumor set. The high p16/low RB expression in TN tumors is consistent with disruption of the RB pathway (3) and suggest these TN tumors may depend on miR17-92 to down-modulate expression of E2F1 which would otherwise induce apoptosis. Conclusions: Our results show that miRNA expression in breast cancer tumors differs depending on subtype. We also find evidence to suggest subtype-specific miRNA regulation of proliferation and a particular dependency of TN tumors on the miR17-92 cluster for survival.

Retrospective Analysis of Thrombocytopenia in Relapsed Multiple Myeloma Patients

Abstract 5073 IntroductionDespite improvement in patient (pt) outcomes in recent years, multiple myeloma (MM) remains incurable and nearly all pts relapse after initial response to therapy. With disease progression and cumulative exposure to chemotherapeutic agents, pts with relapsed MM frequently develop cytopenias that represent a significant clinical challenge. At present, management of high-grade thrombocytopenia at our institution consists of supportive platelet transfusions. We hypothesize that thrombocytopenia is a highly relevant variable in the overall management of relapsed MM. As there are currently no studies in the myeloma literature that assess the impact of thrombocytopenia, we conducted a retrospective, chart-review study to evaluate the impact of thrombocytopenia in relapsed MM. MethodsParticipants included all pts with relapsed and/or relapsed-refractory MM who participated in clinical trials at Dana Farber Cancer Institute from January 1, 2007 – December 31, 2009. Many pts participated in more than one clinical trial during this time period. In this analysis, therapy administered to a particular patient in the context of one clinical trial is referred to as a “treatment regimen.” The overall incidence of thrombocytopenia (platelet count < 150,000/mcl) at initiation of a treatment regimen was recorded. The incidence of severe thrombocytopenia (platelet count < 100,000/mcl and < 75,000/mcl) was also determined. Medical record data collected to assess the clinical impact of thrombocytopenia included the following variables: episodes of ≥ grade 3 thrombocytopenia, number of platelet transfusions per pt per month on clinical trial, treatment delays and treatment discontinuations due to thrombocytopenia, the occurrence of bleeding events, and overall survival on clinical trial therapy. Point estimates with exact binomial confidence intervals and median and range are used to describe the data. The differences in platelet transfusion requirements were assessed by the two-sided Wilcoxon rank sum test. ResultsTo date, data has been abstracted on 43 treatment regimens in 23 pts. Fifteen pts had previously undergone autologous stem cell transplantation (ASCT), including two who had undergone tandem auto-transplant. The overall incidence of thrombocytopenia (platelet count < 150,000/mcl) was 40% at time of treatment initiation. In 13 of the 43 (30%, 90% Confidence Interval (CI)) treatment regimens, the platelet count at time of treatment initiation was less than 100,000/mcl. The platelet count at time of treatment initiation was less than 75,000/mcl in 11 of 43 (25%, 90%CI) treatment regimens reviewed. Cases where the platelet count was ≤ 100,000/mcl at time of study entry, mean platelet transfusion requirement per 4-week time interval was 2.35 units, while in cases where the platelet count was ≥ 100,000/mcl, the mean platelet transfusion requirement was 0.20 units transfusions per 4 week period was needed (p <.0001). In 20 (47%, 90% CI) of the treatment regimens reviewed, bortezomib was part of the therapeutic regimen. Data revealed no significant difference in requirement for platelet transfusion between regimens that included bortezomib versus those that did not. Two pts experienced a bleeding event during their treatment regimens. The platelet count at the time of the bleeding events was 57,000/mcl and 154,000/mcl. Treatment was delayed for one pt due to thrombocytopenia, and two pts were removed from study due to thrombocytopenia. Overall rate of survival at 1-year among 23 pts assessed to date was 91%. ConclusionThrombocytopenia is a common problem in this pt population, particularly in pts that begin treatment with a platelet count < 100,000/mcl. This results in increased platelet transfusions, which are associated with heightened cost of health care and risk of transfusion related toxicities (ex. platelet alloimmunization). Complications related to thrombocytopenia such as bleeding events and treatment discontinuation are relatively rare; this is likely due to routine use of platelet transfusion. Of note, the incidence of transfusion is likely underrepresented by this retrospective study because intact bone marrow function is an eligibility criterion for most clinical trials involving pts with relapsed MM. This data supports the need for improved therapies that minimize transfusion support in this population. Disclosures:Richardson:Millennium:; Celgene:; Johnson & Johnson:; Novartis:; Bristol Myers Squibb:. Anderson:Celgene: Consultancy, Honoraria; Millennium Pharmaceuticals, Inc.: Consultancy, Honoraria; Novartis: Consultancy, Honoraria.

Magnetic resonance (MR) perfusion imaging to differentiate early progression from pseudoprogression following chemoradiotherapy for glioblastoma (GBM).

2009 Background: Patients with glioblastoma (GBM) treated with temozolomide and radiotherapy (RT) may experience early progressive disease (PD) or pseudoprogression (PsP) due to radiation effects, both of which appear as contrast-enhancing lesions on MRI. Currently, there is no reliable way to differentiate between PD and PsP, posing a common diagnostic and treatment dilemma since patients with PD have a poor prognosis, while PsP usually resolves without changes in therapy. Methods: We retrospectively identified 71 GBM patients with new or enlarging enhancing lesions on initial MRI following temozolomide and RT. In twenty patients, dynamic susceptibility (DSC) MR perfusion had been performed, including relative cerebral blood volume (rCBV), relative peak height (rPH) and peak signal recovery (PSR) of the enhancing lesions. Two independent reviewers unaware of results of DSC MR perfusion imaging assigned patients to either PD or PsP based on review of clinical data and follow up or results of biopsy of the enhancing lesions (if available). Two-sided Wilcoxon rank-sum tests were used to evaluate associations between DSC parameters and PD or PsP status. Results: The PsP group had significantly lower median rCBV compared to PD (1.5 vs 2.8, p=0.009) and rPH (1.3 vs 3.0, p<0.001), with higher PSR (1.0 vs 0.8, p=0.039). Receiver operator characteristic analysis revealed an optimal rCBV threshold of ≤1.80 to detect PsP with 100% sensitivity and 75% specificity (AUC=0.938). To maximize specificity, an rCBV threshold of ≤2.40 achieved 69% sensitivity and 100% specificity to detect PsP. Conclusions: DSC MR perfusion imaging of new or enlarging enhancing lesions immediately after completion of RT and temozolomide successfully identified patients with PsP versus those with PD, which has important implications for clinical treatment.

Safety of anti‐IgE treatment with omalizumab in children with seasonal allergic rhinitis undergoing specific immunotherapy simultaneously

Seasonal allergic rhinitis (SAR) affects at least 10-25% of the Caucasian race and about 40% of patients are children. Standard treatment of SAR is specific immunotherapy (SIT), but anti-allergic drugs can significantly enhance efficacy of SIT. One candidate is the humanized monoclonal anti-IgE antibody omalizumab. Randomized, double-blind, placebo-controlled, multi-centre trial in Germany. A total of 221 children were randomly assigned to four different groups and treated with SIT (either grass or birch pollen), starting at least 14 wk before the local birch pollen season. After the 12-wk SIT titration phase, either anti-IgE (omalizumab) or placebo (NaCl 0.9%) therapy was added. This combination therapy with SIT and anti-IgE or placebo lasted 24 wk. To record local reactions and adverse events (AE), the injection site was examined by a clinician 20 min after application of study medication. Further, patients stated any AE and the use of rescue medication by means of a diary 3 days after every injection. Finally, any AE or serious adverse event (SAE) reported by the patients was specified on a standard form by clinicians. Overall tolerance was judged by the doctors according to the patient's diaries. To test differences between the groups, we used either the two-sided Wilcoxon rank-sum test or the two-sided chi-square test. Tolerability of SIT and omalizumab treatment was good (82% of patients). Only some AE with possible causal relationship to treatment occurred slightly more often in the verum groups, i.e. local reactions (16.8 vs. 12.3%) and gastrointestinal (2.7 vs. 0.9%) and ear symptoms (1.8 vs. 0%). Most AE (93.4% in omalizumab and 87.2% in placebo group) were judged by the patients as mild to moderate. SAE were restricted to four patients with asthma in the placebo group, two subjects with headache in the verum group and three patients with infections (two in verum and one in placebo group). Only the cases of asthma were judged to be possibly related to study medication. Further, redness and swelling at the SIT injection site appeared significantly more often in the placebo group which suggests a positive effect of omalizumab on local reaction induced by SIT. Omalizumab represents an important clinical advance in the management of allergic diseases and can be considered to be safe in children.