Dunn's Multiple Comparison Test Research Articles

Analysis of hard tissue regeneration and Wnt signalling in dental pulp tissues after direct pulp capping with different materials.

To investigate the involvement of Wnt signalling and cell cycle regulation in hard tissue formation after pulp capping with several materials in a rat molar pinpoint exposure model. Thirty-two rat molar pulps were mechanically exposed and assigned to 4 groups according to the pulp capping materials used Ca(OH)2 , mineral trioxide aggregate (MTA), Biodentine™ and an untreated control group. After 4weeks, the teeth were collected for microcomputed tomography to quantify reparative dentine formation. Histological analysis was then performed to evaluate the quality of the reparative dentine and the dental pulp tissue inflammatory reaction. Cyclin D1 and β-catenin expression was examined using immunofluorescence staining. The Kruskal-Wallis followed by Dunn's multiple comparison test was performed to determine significant differences. The exposed dental pulps treated with Ca(OH)2 , MTA and Biodentine™ exhibited reparative dentine formation near the exposure site. Fibrous tissues adjacent to the exposure site were observed in the untreated group. The microcomputed tomography evaluation of MTA and Biodentine™ groups revealed significantly greater reparative dentine formation compared with the control group (P=0.0032 in the MTA group and P=0.05 in the BiodentineTM group). From histological evaluations, the BiodentineTM group exhibited significantly greater reparative dentine formation grading compared with the control group (P=0.0152). The pulp tissues treated with Ca(OH)2 and Biodentine™ exhibited a lower inflammatory score compared with those of the untreated control (P=0.0291 in the Ca(OH)2 and P=0.0110 in the BiodentineTM group). Ca(OH)2 , MTA and Biodentine™ induced cyclin D1 expression in the dental pulp tissues adjacent to the reparative dentine. Moreover, the Biodentine™-treated defects demonstrated β-catenin expression in the pulp tissue adjacent to the newly formed reparative dentine, which was not observed with the other materials. All test materials promoted dentine bridge formation and stimulated cyclin D1 expression. The favourable outcome after direct pulp capping with Biodentine™ involved Wnt/β-catenin signalling. However, Wnt/β-catenin signalling did not participate in the mechanism by which Ca(OH)2 and MTA promoted reparative dentine formation.

The effects of sodium hypochlorite and ethylenediaminetetraacetic acid on the microhardness of Mineral Trioxide Aggregate and TotalFill Bioceramic Putty.

This study aimed to evaluate the effects of 5% NaOCl and 17% EDTA on the Vickers microhardness of Mineral Trioxide Aggregate, MTA Angelus (MTAA), and TotalFill Bioceramic Putty (BCP) after 24h and 8days. Twenty-four samples of MTAA and BCP were tested for baseline microhardness at 24h. They were divided into four subgroups (5% NaOCl or 17% EDTA, 24h, 5% NaOCl or 17% EDTA at 8days) and microhardness was evaluated at different time-points. Results were recorded and analysed statistically via one-way anova Kruskal-Wallis test and post hoc uncorrected Dunn's multiple comparison test. MTAA had a higher baseline microhardness than BCP. NaOCl reduced the microhardness of MTAA but increased that of BCP. EDTA reduced the microhardness for both materials at 24h. At 8days, EDTA reduced the microhardness of MTAA, while BCP exhibited an increase in microhardness. Enabling these materials to set for a week prior to exposure to irrigants improves their microhardness.

Preliminary results on the association with feeding and recovery length in equine colic patients after laparotomy.

Colic is a serious disease for horses and the nutritional management of postoperative colic patients is an extremely important field. The aim of this retrospective study was to analyse the different factors, especially related to nutritional management, that may be associated with recovery length during hospitalization after a surgical intervention for colic, using a multivariate model. Data were collected from the records of horses presented to two hospitals and undergoing surgery for colic. The length (days) of recovery was the outcome of interest and was taken into account as a reference parameter (short, medium, long). The parameters collected (patient details, preoperative clinical and laboratory examinations, postoperative parameters and post-surgery nutritional parameters) were subjected to multivariate analysis (MCA and PCoA). A ranking class dataset was used to calculate Kendall's tau correlation of the length of recovery with respect to other parameters. Descriptive statistic to identify differences in the recovery length among groups (Kruskal-Wallis and Dunn's Multiple Comparison Test) was also performed. p value was set at<0.05. Groups were not different in preoperative clinical parameters (BCS, PCV, total protein), postoperative parameters (time to 1st defecation, time to the end of IV fluid therapy and time to first water drinking, anaesthesia) and patient details (age). The comparison among groups revealed differences on the post-surgical nutritional parameters. Horses with short recovery consumed higher % of DM as forages in the 24hr compared with the horses that have a long recovery and reach the minimum DM intake in a shorter period. Both Kendall and MCA analysis confirmed that the time to first feeding had a positive association with the length of recovery. Only 37 horses undergoing colic surgery were included in the study. From a clinical standpoint, this study has shown which nutritional parameters are associated with short recovery.

Kinematics of a novel reciprocating endodontic handpiece.

To analyse the kinematics of the Reciproc Direct® contra-angle reciprocating device with different motor sources. Reciproc Direct contra-angle (VDW GmbH, Munich, Germany) was tested with new micro-motor sources. The micro-motor groups were as follows: a brushless electric micro-motor, a brushed electric micro-motor and an air-driven micro-motor. The electric micro-motor sources were also divided intofour subgroups as 10000, 15000, 20000 and 25000rpm. The maximum air pressure of the air-driven micro-motor was adjusted to 2 and 3kgfcm-2 . A custom target object was attached to the Reciproc Direct, and reciprocating motions were recorded with a high-speed camera at 1200frames per second. The following kinematic parameters were calculated: duration of each reciprocating motion, engaging and disengaging angles, cycle rotational speeds, engaging and disengaging rotational speeds, net cycle angle, total cycle angle and number of cycles to complete full rotation. One-way anova was used where applicable, followed by Tukey's multiple comparison tests, to compare the kinematic values of reciprocating motion for each micro-motor/Reciproc Direct combination. The Kruskal-Wallis test followed by Dunn's multiple comparison test was used for non-normally distributed data. Statistical analysis was performed (α=0.05). For the brushless micro-motor, median engaging angle was 186.5° at 10000rpm and 188.0° at 15000rpm which were significantly different than median engaging angles at 20000 (188.5°) and 25000 (189.3°) rpm (P<0.05). For the brushless micro-motor, median cycle rotational speed was 372.5rpm at 10000 and 459.8rpm at 15000rpm which were significantly different than median cycle rotational speed at 20000 (576.2rpm) and 25000 (677.8rpm) rpm (P<0.05). For the brushed micro-motor, median cycle rotational speed was 293.5rpm at 10000 and 386.3rpm at 15000rpm which were significantly different than median cycle rotational speed at 20000 (508.9rpm) and 25000 (597.6rpm) rpm (P<0.05). Rotational speeds were influenced significantly by motor sources even when the Reciproc Direct was used at speeds recommendedby the manufacturer. This could indicate that thekinematics of the Reciproc Direct are dependent on the power of the rotating motor.

Cerebrospinal fluid BDNF pro-peptide levels in major depressive disorder and schizophrenia

The role of brain-derived neurotrophic factor (BDNF) and its related molecules has been extensively studied in the context of psychiatric disorders. In the present study, we focused on the newly identified BDNF pro-peptide, which is generated together with mature BDNF by proteolytic processing of their precursor, proBDNF. Here, we report, for the first time, that BDNF pro-peptide is present in human cerebrospinal fluid (CSF) and quantifiable by western blotting. We measured CSF BDNF pro-peptide levels in 27 patients with schizophrenia, 18 patients with major depressive disorder (MDD), and 27 healthy controls matched for age, sex, and ethnicity (Japanese). The ratio of the BDNF pro-peptide level to the total protein level in MDD patients was significantly lower than that in controls (Kruskal-Wallis with Dunn's multiple comparisons test; p = 0.046). When men and women were examined separately, males with MDD had a significantly lower BDNF pro-peptide/protein ratio than male controls (p = 0.047); this difference was not found in female subjects. The ratio tended to be lower in male schizophrenia patients (p = 0.10). Although we tried to measure the levels of mature BDNF in CSF, they were below the limit of detection of the ELISA and multiple analyte profiling technology. Taken together, the results suggest that reduced CSF BDNF pro-peptide levels are associated with MDD, particularly in males. Further studies involving a larger sample size are warranted.

Efficacy of reciprocating systems for removing root filling material plus complementary cleaning methods in flattened canals: Microtomography and scanning electron microscopy study.

The aim of this study was to evaluate three reciprocating systems and complementary cleaning methods on filling material removal during retreatment of flattened canals. Thirty-six mandibular incisors were prepared using rotary instruments up to size 25.08 and filled using the single-cone technique. Subsequently, the teeth were divided into three groups (n = 12) according to retreatment procedures: Reciproc Blue (RB): 25/0.08 and 40/0.06; ProDesign R (PDR): 25/0.06 and 35/0.05; and WaveOne Gold (WOG): 25/0.07 and 35/0.06. The remaining filling volume materials were assessed by means of micro-CT imaging before and after retreatment. After this, the specimens were subdivided into three groups according to complementary cleaning methods: XP-Endo Shaper (30/0.01); passive ultrasonic irrigation (PUI); 60° oscillatory instrumentation with #30 H-file, and micro-CT scan was taken. Then, the roots were cut in half and the samples were analyzed by scanning electron microscope (SEM). The data were analyzed statistically using Kruskal-Wallis and Dunn's Multiple Comparison test with significance level of 5%. None of the reciprocating files promoted complete removal of filling material and there was no statistical difference between the groups, regardless instrument size (p > 0.05). Complementary cleaning methods increased remnant filling removal (p < 0.05). XP-endo Shaper significantly reduced the amount of filling material in the apical and middle thirds, compared with H-files (p < 0.05), with no difference with PUI. In the SEM, there was no statistical difference among the instruments (p > 0.05). The reciprocating systems showed similar effectiveness in removing root filling material. Complementary cleaning method with the XP-Shaper enhanced filling material removal.

CLAD Phenotypes Determined by Change in Spirometric Index Associate with Quantitative Image Analyses

Purpose Accurate phenotypes of chronic lung allograft dysfunction (CLAD) are critical to study disease mechanisms and ensure homogenous populations for clinical trials, but the optimal classification method is unknown. A restrictive CLAD phenotype involves parenchymal disease similar to pulmonary fibrosis. The Center for Computer Vision and Imaging Biomarkers at UCLA has established CT based computer aided diagnosis (CAD) scores for pulmonary fibrosis. We applied CT CAD scores to CLAD and compared 2 previously described methods for CLAD classification. Methods The cohort includes 73 lung recipients; 44 no CLAD and 29 with a CT done within 90 days of CLAD onset. The FVC loss method defines restrictive-CLAD (RCLAD) as a ≥20% decline in FVC from baseline at CLAD onset. The spirometric change method uses relative change in FVC divided by relative change in FEV1 from baseline to CLAD onset, and a value >0.5 is classified as restrictive allograft syndrome (RAS). For both methods, when RCLAD or RAS is not established, the default is bronchiolitis obliterans syndrome (BOS). We compared the quantitative CT CAD scores for QLF (fibrosis), QGG (ground glass), and QILD (QLF + QGG), expressed as a percentage of total lung involvement at TLC, between CLAD phenotypes using Dunn's multiple comparisons tests. Results Of the 29 CLAD cases, 15 were classified as RCLAD and 19 as RAS. The classification methods agreed in 21 cases: 13 RCLAD/RAS and 8 BOS/BOS. The methods disagreed in 8 cases: 2 RCLAD/BOS and 6 BOS/RAS. RCLAD was not associated with differences in QLF, QGG, or QILD. However, RAS was associated with significantly increased QLF, QGG, and QILD scores as compared to BOS and to no CLAD (Figure). Conclusion The spirometric change method for CLAD phenotype classification is associated with radiographic parenchymal lung disease, suggesting it is a better method to distinguish CLAD phenotypes. Our study is limited by small sample size and we have not yet incorporated features of air trapping from RV series.

An investigation of the effect of fat suppression and dimensionality on the accuracy of breast MRI segmentation using U-nets.

Accurate segmentation of the breast is required for breast density estimation and the assessment of background parenchymal enhancement, both of which have been shown to be related to breast cancer risk. The MRI breast segmentation task is challenging, and recent work has demonstrated that convolutional neural networks perform well for this task. In this study, we have investigated the performance of several two-dimensional (2D) U-Net and three-dimensional (3D) U-Net configurations using both fat-suppressed and nonfat-suppressed images. We have also assessed the effect of changing the number and quality of the ground truth segmentations. We designed eight studies to investigate the effect of input types and the dimensionality of the U-Net operations for the breast MRI segmentation. Our training data contained 70 whole breast volumes of T1-weighted sequences without fat suppression (WOFS) and with fat suppression (FS). For each subject, we registered the WOFS and FS volumes together before manually segmenting the breast to generate ground truth. We compared four different input types to the U-nets: WOFS, FS, MIXED (WOFS and FS images treated as separate samples), and MULTI (WOFS and FS images combined into a single multichannel image). We trained 2D U-Nets and 3D U-Nets with these data, which resulted in our eight studies (2D-WOFS, 3D-WOFS, 2D-FS, 3D-FS, 2D-MIXED, 3D-MIXED, 2D-MULTI, and 3D-MULT). For each of these studies, we performed a systematic grid search to tune the hyperparameters of the U-Nets. A separate validation set with 15 whole breast volumes was used for hyperparameter tuning. We performed Kruskal-Walis test on the results of our hyperparameter tuning and did not find a statistically significant difference in the ten top models of each study. For this reason, we chose the best model as the model with the highest mean dice similarity coefficient (DSC) value on the validation set. The reported test results are the results of the top model of each study on our test set which contained 19 whole breast volumes annotated by three readers fused with the STAPLE algorithm. We also investigated the effect of the quality of the training annotations and the number of training samples for this task. The study with the highest average DSC result was 3D-MULTI with 0.96±0.02. The second highest average is 2D WOFS (0.96±0.03), and the third is 2D MULTI (0.96±0.03). We performed the Kruskal-Wallis one-way ANOVA test with Dunn's multiple comparison tests using Bonferroni P-value correction on the results of the selected model of each study and found that 3D-MULTI, 2D-MULTI, 3D-WOFS, 2D-WOFS, 2D-FS, and 3D-FS were not statistically different in their distributions, which indicates that comparable results could be obtained in fat-suppressed and nonfat-suppressed volumes and that there is no significant difference between the 3D and 2D approach. Our results also suggested that the networks trained on single sequence images or multiple sequence images organized in multichannel images perform better than the models trained on a mixture of volumes from different sequences. Our investigation of the size of the training set revealed that training a U-Net in this domain only requires a modest amount of training data and results obtained with 49 and 70 training datasets were not significantly different. To summarize, we investigated the use of 2D U-Nets and 3D U-Nets for breast volume segmentation in T1 fat-suppressed and without fat-suppressed volumes. Although our highest score was obtained in the 3D MULTI study, when we took advantage of information in both fat-suppressed and nonfat-suppressed volumes and their 3D structure, all of the methods we explored gave accurate segmentations with an average DSC on >94% demonstrating that the U-Net is a robust segmentation method for breast MRI volumes.

The Use of Two-Dimensional Strain Measured by Speckle Tracking in the Identification of Incipient Ventricular Dysfunction in HIV-Infected Patients on Antiretroviral Therapy, Untreated HIV Patients and Healthy Controls.

BackgroundMost cardiovascular abnormalities in patients infected with the human immunodeficiency virus (HIV) have been associated with myocardial damage directly caused by the virus. Some cases, however, may be associated with adverse effects from antiretroviral therapy (ART). New ventricular function assessment techniques are capable of detecting early changes in the cardiac function of HIV-infected patients using or not using ART. The usefulness of these techniques has been little employed in these patients.ObjectivesTo investigate the potential influence of antiretroviral therapy (ART) on the occurrence of subclinical left ventricular systolic dysfunction evaluated by myocardial strain rate analysis using two-dimensional speckle tracking echocardiography (2-D Echo) in treated HIV patients compared to untreated patients and healthy individuals.MethodsSixty-eight HIV-infected patients with no cardiovascular symptoms, normal left ventricular (LV) ejection fraction (> 0.55 on 2-D Echo) were divided into three groups: 11 patients not using antiretroviral therapy (NT), 24 using protease inhibitor (PI) and 33 using non-nucleoside reverse transcriptase inhibitor (NNRTI). We also studied 30 normal non-HIV infected individuals (Ctrl). Demographic, clinical, biochemical and anthropometric data were collected. Preliminary transthoracic echocardiography included study of myocardial strain using two-dimensional speckle tracking. We studied strain and strain rate in the seventeen left ventricular (LV) myocardial segments in the longitudinal, circumferential and radial axes. Statistical analysis of the data was done with IBM SPSS - version 20 for Windows. Upon analysis of the data, namely the normality of independent variables in the different groups and the homogeneity of the variances between the groups, Kruskal-Wallis’ non-parametric test was done, followed by Dunn’s multiple comparison tests to test the significance of the differences between the values measured in the study groups. A significance level of 5% was adopted for decision-making on statistical tests.ResultsThe mean age of HIV patients was 40 ± 8.65 years and the mean age of controls was 50 ± 11.6 years (p < 0.001). Median LV global longitudinal strain (GLS) of NT patients (-17.70%), PI patients (-18.27%) and NNRTIs (-18.47%) were significantly lower than that of the Ctrl group (-20.77%; p = 0.001). There was no significant difference in mean SLG between treated patients (PI, NNRTI) and untreated (NT) patients. No significant differences were observed in mean circumferential and radial strain, nor on circumferential and radial strain rates between the NT, PI, NNRTI and Ctrl groups.ConclusionThe data suggest that HIV patients present, on myocardial strain measured by speckle tracking, signs of early LV systolic dysfunction that seem to be unrelated to the presence of ART. The prognostic significance of this condition in these patients deserves further studies.

Comparative evaluation of microleakage around Class V cavities restored with alkasite restorative material with and without bonding agent and flowable composite resin: An in vitro study

Marginal adaptability of restorative material is one of the prime factors for success of a restoration. To evaluate microleakage at enamel restoration and dentin restoration interface of Class V cavities restored with new alkasite restorative material Cention-N, with and without using bonding agent and flowable composite resin. Thirty Class V tooth preparations were divided into three groups (n = 10): Group-I restored with Cention-N (Ivoclar Vivadent) without adhesive, Group-II was restored with Cention-N after application of eighth-generation bonding agent (3M ESPE, Single Bond Universal Adhesive), and Group-III was restored with flowable composite resin (Tetric-N-Flow, Ivoclar Vivadent). All samples were subjected to 200 thermocycles between temperature baths at 5°C and 55°C. All samples were cut longitudinally through the center of the restorations with the help of isomet diamond saw. The sections were then observed under binocular stereomicroscope at 20×. Two evaluators scored the depth of dye penetration independently at enamel and dentin margins. Kruskal-Wallis nonparametric analysis followed by Dunn's multiple comparison tests were done to evaluate differences among the experimental groups. Mann-Whitney test was used to compare the difference between occlusal and gingival scores within each restoration. Microleakage seen in decreasing order: Cention-N without adhesive >Flowable composite >Cention-N with adhesive. Microleakage at enamel restoration interface was less than microleakage at dentin restoration interface of each group, but the difference was not statistically significant. Least microleakage was seen with Cention-N with adhesive followed by flowable composite. More microleakage was seen with Cention-N without adhesive.

The ability of three nickel-titanium mechanized systems to negotiate and shape MB2 canals in extracted maxillary first molars: a micro-computed tomographic study.

To evaluate the influence of three engine-driven NiTi file systems manufactured from different NiTi alloys for the preparation of second mesiobuccal (MB2) canals in extracted maxillary first molars using a micro-computed tomographic (micro-CT) scanner. Thirty maxillary molars with two canals in the mesiobuccal root were selected and randomly divided into three groups (n=10): Reciproc [REC; size 25, .08 taper; VDW, Munich, Germany], ProDesign R [PDR; size 25, .06 taper; Easy, Belo Horizonte, Brazil] and Mtwo [MO; size 25, .06 taper; VDW, Munich, Germany]. Before and after root canal preparation of the MB2 canal, the teeth were scanned using a micro-CT to evaluate canal transportation, centring ability, dentine thickness and volume change. The working time to achieve working length was also evaluated. All parameters were compared statistically using the Kruskal-Wallis and Dunn test for multiple comparisons, with a significance level of 5%. There was no significant difference amongst the groups regarding canal transportation and centring ability (P>0.05). However, the PDR size 25, .06 taper group had significantly lower canal volume and volume of dentine removal compared with a MO size 25, .06 taper and REC size 25, .08 taper (P<0.05). A root perforation was detected in MO size 25, .06 taper and REC size 25, .08 taper groups, respectively. Regarding the working time, the PDR size 25, .06 taper required a significantly longer time to achieve working length than MO size 25, .06 taper and REC size 25, .08 taper (P<0.05). All NiTi systems had similar canal transportation, centring ability and increase in apical volume after preparation of MB2 canals. However, the PDR size 25, .06 taper had less volume of dentine removal, absence of root canal perforation and required a longer time to accomplish the root canal preparation.

Longitudinal airway remodeling in active and past smokers in a lung cancer screening population.

To longitudinally investigate smoking cessation-related changes of quantitative computed tomography (QCT)-based airway metrics in a group of heavy smokers. CT scans were acquired in a lung cancer screening population over 4years at 12-month intervals in 284 long-term ex-smokers (ES), 405 continuously active smokers (CS), and 31 subjects who quitted smoking within 2years after baseline CT (recent quitters, RQ). Total diameter (TD), lumen area (LA), and wall percentage (WP) of 1st-8th generation airways were computed using airway analysis software. Inter-group comparison was performed using Mann-Whitney U test or Student's t test (two groups), and ANOVA or ANOVA on ranks with Dunn's multiple comparison test (more than two groups), while Fisher's exact test or chi-squared test was used for categorical data. Multiple linear regression was used for multivariable analysis. At any time, TD and LA were significantly higher in ES than CS, for example, in 5th-8th generation airways at baseline with 6.24mm vs. 5.93mm (p < 0.001) and 15.23mm2 vs. 13.51mm2 (p < 0.001), respectively. RQ showed higher TD (6.15mm vs. 5.93mm, n.s.) and significantly higher LA (14.77mm2 vs. 13.51mm2, p < 0.001) than CS after 3years, and after 4years. In multivariate analyses, smoking status independently predicted TD, LA, and WP at baseline, at 3years and 4years (p < 0.01-0.001), with stronger impact than pack years. Bronchial dimensions depend on the smoking status. Smoking-induced airway remodeling can be partially reversible after smoking cessation even in long-term heavy smokers. Therefore, QCT-based airway metrics in clinical trials should consider the current smoking status besides pack years. • Airway lumen and diameter are decreased in active smokers compared to ex-smokers, and there is a trend towards increased airway wall thickness in active smokers. • Smoking-related airway changes improve within 2years after smoking cessation. • Smoking status is an independent predictor of airway dimensions.

Prospects for early childhood feeding interventions: An exploration of parent's concerns and acceptability towards social media intervention opportunities.

The present study aims to (i) consult with parents regarding child feeding concerns, as relevant to child feeding interventions, (ii) determine parents' willingness to participate in online and social media-based interventions and (iii) determine differences in intervention acceptability based on geographic diversity and demographic characteristics. Three hundred thirty Australian parents of children (2.0-5.0 years) with child feeding concerns participated in an online, cross-sectional survey. Kruskal-Wallis with Dunn's multiple-comparison test, using Bonferroni adjustment, was used to compare differences between variables and participants characteristics (region of residence, parent age, parent education and income). About 53.9% of respondents were concerned about child fussy eating, with lack of time and child tantrums common barriers to addressing concerns. Respondents indicated that a combination of online platforms (websites, email, Facebook) was their preferred method of intervention participation, although, a combination of online and face-to-face methods also had modest preference, particularly among lower educated parents. Participants indicated that they would participate in an online intervention (participating once a week) for more than 12 weeks (42.7%), compared with only 4 weeks (participating once a week) for traditionally delivered interventions. About 32.4% of respondents said they would be concerned about their privacy in an online intervention. Fussy eating is likely to be an appealing focus for future child feeding interventions, with strategies to address common barriers an important curriculum area. Online interventions are likely to offer an acceptable and plausible alternative to traditional interventions that are able to reach geographically diverse populations, although face-to-face intervention components still hold appeal.

Assessment of Emicizumab-Driven Clot Stability in Hemophilia a Model

Clot stability is an important hemostatic aspect in treating patients with hemophilia A (HA). Effects of factor VIII (FVIII), recombinant activated FVII (rFVIIa), FXIII and tranexamic acid on clot stability in hemophilia had been reported1). Emicizumab (Emi), FVIIIa-mimicking bispecific antibody, is a recently approved novel therapeutics for HA with inhibitor. Although hemostatic activity of Emi was confirmed by preclinical and clinical studies, whether Emi-driven clot stability is similar to that of FVIII or not remains unknown. Furthermore, in clinical trial of Emi, thrombotic events were reported when high doses of activated prothrombin complex concentrates (aPCC) were used concurrently with Emi. Effect of aPCC on clot stability and its possible association with the thrombotic events should be also clarified. In this study, we investigated the effect of Emi on clot stability with a thromboelastometry (ROTEM) -based assay using tissue plasminogen activator (tPA) as a fibrinolysis trigger, and compared it with that of FVIII or bypassing agents (BPAs). Furthermore, we assessed the effect of BPAs on Emi-driven clot stability.Normal blood from healthy volunteer (n=22) was incubated with anti-FVIII antibody (FVIII:C <1 IU/dL), which was regarded as HA model blood. HA model blood was spiked with Emi (50 μg/ml), rFVIIa (22 nM), and aPCC (1 U/ml). ROTEM was performed with/without tPA (2 nM). We used ellagic acid (Elg, x300 INTEM®) as a trigger in addition to tissue factor (TF, 0.5 pM) since TF-trigger was not sensitive to Emi. Residual area under the curve in 30 min (%AR30) was used as a clot stability parameter. Median values (25-75%tile) of %AR30 were compared among the groups with Dunn's multiple comparison test [*p<0.05, **p<0.01, ***p<0.001].In Elg-trigger ROTEM, %AR30 of HA model blood had no values since coagulation did not occur within test time. However, coagulation was improved in HA model spiked with Emi and its clot stability showed slightly weak value with no statistical significance [50% (30-76), p>0.05] compared to normal blood [75% (56-90)]. Whilst, HA model + rFVIIa [13%*** (7-22)] and HA model + aPCC [41%*** (20-71)] showed significantly weak values, indicating that effect of Emi on clot stability was comparable to normal blood and better than BPAs in Elg trigger. Concomitant effect of Emi and BPAs were evaluated. Compared to HA model + Emi [50% (30-76)], clot stability increased in the co-presence of Emi and aPCC [99%*** (96-101)] but not in that of Emi and rFVIIa [53% (29-78)].TF trigger, which was sensitive to BPAs but not to Emi, was also used in order to evaluate concomitant effect of Emi and BPAs. In TF trigger, clot stability of HA model blood + Emi [31% (19-47)] was similar to that of HA mode blood [27% (20-44)] or normal blood [32% (22-55)], but increased in HA model + rFVIIa [43%** (30-62)] and HA model + aPCC [41%** (29-68)]. rFVIIa increased clot stability of HA model blood in the presence of Emi [56%** (28-65)] more than its absence. Effect of the aPCC in the presence of Emi [87%*** (78-91)] was much more remarkable than that of rFVIIa.In conclusion, effect of Emi on clot stability in HA model blood was comparable to that of normal blood, indicating that clot formation under Emi had no qualitative difference with FVIII. Clot stability remarkably increased in Emi in the presence of aPCC, suggesting that hyper clot stability might be one of the reasons of thrombotic events in the concomitant use of Emi and aPCC.1) Tran HT, et al. Tranexamic acid as adjunct therapy to bypassing agents in haemophilia A patients with inhibitors. Haemophilia. 2014 May;20(3):369-75. DisclosuresOgiwara:CSL Behring: Research Funding. Nogami:Chugai Pharmaceutical Co., Ltd: Consultancy, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties: Anti-FIXa/X bispecific antibodies , Research Funding, Speakers Bureau. Kitazawa:Chugai Pharmaceutical Co., Ltd: Employment, Equity Ownership, Patents & Royalties: Patents related to emicizumab. Shima:F. Hoffmann-La Roche Ltd: Membership on an entity's Board of Directors or advisory committees; Chugai Pharmaceutical Co., Ltd: Consultancy, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties: Anti-FIXa/X bispecific antibodies , Research Funding, Speakers Bureau.

Identification of Levels of Serum Amyloid A and Apolipoprotein A1 in Serum Proteomic Analysis of Neuropsychiatric Systemic Lupus Erythematosus Patients.

Neuropsychiatric Systemic Lupus Erythematosus (NPSLE) has multiple pathogenic mechanisms that cause diverse manifestations and whose diagnosis is challenging because of the absence of appropriate diagnostic tests. In the present study the application of proteomics using two-dimensional electrophoresis (2D) and mass spectrometry (MS) allowed the comparison of the protein profile of the serum low and high abundance protein fractions of NPSLE patients (NPSLE group) and SLE without neuropsychiatric syndromes (SLE group), Neuropsychiatric syndromes not associated with SLE (NPnoSLE groups), and healthy controls (CTRL group). The gels obtained were digitalized and analyzed with the PDQuest software. The statistical analysis of the spots was performed using the nonparametric Kruskal Wallis and Dunn's multiple comparison tests. Two spots showed significant differences and were identified by MS. Spot 4009 was significantly lower in NPSLE with regard to NPnoSLE (p= 0,004) and was identified as apolipoprotein A1 (APOA1) (score 809-1132). Spot 8001 was significantly higher in NPSLE regarding CTRL and NPnoSLE (p= 0,01 y 0,03, respectively) and was identified as serum amyloid A (SAA) (score 725-2488). The proinflammatory high density lipoproteins (HDL) have been described in SLE. In this HDL the decrease of APOA1 is followed by an increase in SAA. This altered level of both proteins may be related to the inflammatory state that is characteristic of an autoimmune disease like SLE, but this is not specific for NPSLE.

Natural regeneration following wind disturbance increases the diversity of managed lowland forests in NE Poland

AbstractQuestionsAre there significant differences in the density and composition of natural regeneration among habitat types? Is the abundance of regeneration higher in patches more seriously damaged by a windstorm than in patches not affected by the wind? Is the species diversity of regeneration greater than the diversity of mature trees prior to disturbance?LocationSzast Protected Forest, NE Poland.MethodsThroughout the Szast P.F., 111 sample plots were distributed in a regular grid 13 years after a windstorm. In plots located in disturbed forests, we measured all the canopy trees and the 30 young individuals of each size class (seedlings, short saplings and tall saplings) closest to the plot centre. In non‐disturbed patches, we measured all trees within a plot of predefined radius. For statistical analyses of differences in the diversity between mature stands and young generation, we used the Kruskal–Wallis test followed by Dunn's multiple comparison test. The relationship between the canopy layer and the young generation of trees was analysed using Spearman's rank correlation and a classic negative binomial regression.ResultsNatural regeneration was more abundant in the coniferous and mixed coniferous habitat types than in the mixed deciduous type. The density of young trees was negatively correlated with the basal area of the trees that survived the windstorm and was positively correlated with canopy tree mortality. After the windstorm, Pinus sylvestris lost more trees than the other species; Picea abies and Betula pendula slightly increased their share; and the species that benefited most from the disturbance was Quercus robur. In the coniferous habitat type, the species diversity of the young generation of trees was higher than the diversity of the canopy trees prior to the windstorm.ConclusionsLeaving wind‐disturbed areas to natural regeneration could be a viable option for converting coniferous plantations into more diverse and species‐rich stands.

Identification of an oncogenic network with prognostic and therapeutic value in prostate cancer.

Identifying critical pathways governing disease progression is essential for accurate prognosis and effective therapy. We developed a broadly applicable and novel systems‐level gene discovery strategy. This approach focused on constitutively active androgen receptor (AR) splice variant‐driven pathways as representative of an intractable mechanism of prostate cancer (PC) therapeutic resistance. We performed a meta‐analysis of human prostate samples using weighted gene co‐expression network analysis combined with experimental AR variant transcriptome analyses. An AR variant‐driven gene module that is upregulated during human PC progression was identified. We filtered this module by identifying genes that functionally interacted with AR variants using a high‐throughput synthetic genetic array screen in Schizosaccharomyces pombe. This strategy identified seven AR variant‐regulated genes that also enhance AR activity and drive cancer progression. Expression of the seven genes predicted poor disease‐free survival in large independent PC patient cohorts. Pharmacologic inhibition of interacting members of the gene set potently and synergistically decreased PC cell proliferation. This unbiased and novel gene discovery strategy identified a clinically relevant, oncogenic, interacting gene hub with strong prognostic and therapeutic potential in PC.

Simultaneous Bilateral Endoscopic Surgery (SBES) for Patients with Bilateral Upper Tract Urolithiasis: Technique and Outcomes

BackgroundThe incidence of bilateral and multiple renal stones is not negligible. To date, some sparse data on simultaneous bilateral stone surgery are available in literature showing good outcomes in terms of both effectiveness and safety. ObjectiveTo describe our series of patients with bilateral renal stones who underwent simultaneous bilateral endoscopic surgery (SBES), reporting its effectiveness and safety. Design, setting, and participantsA prospective analysis of 27 consecutive patients who underwent simultaneous flexible ureteroscopy (fURS) in one side and percutaneous nephrolithotomy (PCNL) in the other side for bilateral renal stones was performed. Surgical procedureSBES, performing fURS in one side and PCNL in the other side contemporaneously. MeasurementsClinical data were collected in a dedicated database. Intra- and postoperative outcomes were assessed. Comparisons among pre- and postoperative serum creatinine levels and estimated glomerular filtration rate values during the study period were performed using the Kruskal-Wallis test with the Dunn multiple comparison test. Results and limitationsAll the procedures were carried out until the end in both sides without encountering any complications intraoperatively. The mean stone size was 27.1±8.1 and 11.1±3.6mm for the PCNL and fURS side respectively. The mean operative time was 79.4±25.2min. There were no differences in patients' creatinine and eGFR when comparing at baseline with 1-mo after SBES. No postoperative major complications were experienced (Clavien-Dindo grade I 3.7%; II 11.1%). Stone-free rate was 74% at 1-mo follow-up. The main limitation of the study is the small size of the group analyzed. ConclusionsSBES is safe and effective, with minimal morbidity. SBES has the potential advantages of shorter operative time, reduced anesthesia, and reduced hospital time, which can benefit patients, surgeons, and health care systems. Patient summarySimultaneous bilateral endoscopic surgery is an effective treatment with low complication rates for bilateral urolithiasis. This innovative and complicated procedure should be performed in high-volume centers by experienced surgeons.

Abstract LB-224: Circulating tumor DNA for early detection and intervention in breast cancer: ctDNA profiles discriminate between healthy women in a true cancer screening setting and disease-free women on follow up

Abstract Successful early detection and intervention in cancer is key if we are to drastically improve cancer survivorship. Previous research by us and others has shown that copy number and mutational profiling of circulating cell-free DNA (cfDNA) can detect minimal residual disease and predict who will relapse after early stage breast cancer (Shaw et al. Genome Res 2012, doi: 10.1101, Garcia-Murillas et al. Sci Transl Med. 2015 doi: 10.1126). Here we used massively parallel sequencing with the Ion CHEF -S5 and Oncomine™ Breast cfDNA Assay (Thermofisher) for ultrasensitive detection of circulating tumour derived DNA (ctDNA) in 26 asymptomatic early stage breast cancer patients with no scan evidence of distant metastases on follow up after surgery, (all of whom were receiving adjuvant AI (PAI) at the time of blood sampling). We compared these early stage breast cancers with cfDNA isolated from 40 patients with metastatic breast cancer (MBC) and 92 healthy age matched female controls (HC) attending for a routine breast screening mammogram. The Oncomine™ Breast cfDNA Assay targets &amp;gt;150 hotspot mutations in 10 breast cancer genes and uses tag sequencing technology to achieve a limit of detection (LOD) down to 0.1%. One or more driver mutations were detected in cfDNA of 19 (73%) PAIs and 29 (72.5 %) MBCs, whereas just 15 HCs (16%) were positive. Applying this threshold of 1 or more driver mutations for detection of ctDNA (Cohen et al. Science. 2018 doi: 10.1126) there was a significant difference between ctDNA positive cancers and healthy controls, which also remained significant for 2 or more driver mutations (P &amp;lt; 0.0001 and P &amp;lt; 0.001 respectively, Fisher's exact test) (Abbosh et al. Nature 2017, doi: 10.1038). It is notable that &amp;gt;72% of breast cancers were positive with this test, both disease-free women on follow up and patients with MBC, suggesting a role for the test in routine monitoring. A second finding of clinical significance was that variant allele fraction of ESR1 mutant ctDNA was significantly higher in cancers than controls (PAIs than HCs (P = 0.034) and MBCs than HCs (P = 0.002); adjusted P value, Dunn's multiple comparison test) suggesting that ESR1 mutations distinguish disease-free breast cancer survivors receiving adjuvant aromatase inhibitors from normal women. Importantly, we are the first to report ultrasensitive evaluation of ctDNA alterations in a large number of healthy women in a true cancer screening setting. The 15 healthy women that were “positive” for ctDNA may be harbouring an early stage cancer not detectable by imaging. Research is ongoing to validate these results in a larger study, but our results suggest that the approach may have utility for early disease detection and monitoring to enable appropriate intervention at earlier stages. Citation Format: Jacqui A. Shaw, Karen Page, Daniel Fernandez-Garcia, Allison Hills, Anna R. Boydell, Lindsay Primrose, Bradley Toghill, Robert K. Hastings, Kelly Gleeson, Brenda M. Rosales, Kate Goddard, David S. Guttery, Simak Ali, Raoul C. Coombes. Circulating tumor DNA for early detection and intervention in breast cancer: ctDNA profiles discriminate between healthy women in a true cancer screening setting and disease-free women on follow up [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-224.

The Effect of Contact Lens Wear on the Cellular Morphology of the Lid Wiper Area.

This cross-sectional study presented a link between contact lens wear and changes on the cellular morphology characteristics of the lid wiper (LW) epithelium, which was not visible by LW staining. The aim of this study was to establish if the duration of contact lens (CL) wear affects the cellular morphology of the LW epithelium. This was a cross-sectional study of 100 individuals with different exposures to CL wear: short, moderate, and long experience of CL wear; previous CL wearers; and nonwearers (NWs) as controls. Impression cytology samples were collected from the central upper lid margin (LW area). After fixing, samples were stained with periodic acid-Schiff and haematoxylin for cell morphology analysis and subsequently graded according to the Nelson 0- to 3-point scale. Lid wiper staining was assessed with the aid of lissamine green and graded using the Korb (0- to 3-point) scale. One-way Kruskal-Wallis analysis followed by the Dunn multiple-comparisons test was used for statistical comparison. The Nelson grade for LW epithelium morphology was significantly different between groups (P = .003). Abnormal epithelial morphology as defined by grade 2 or 3 was evident in 66.7% of CL wearers with short experience and 76.5% of CL wearers with moderate experience. This was significantly higher than NWs of whom only 21.5% showed greater than grade 1 (P = .02 and .005, respectively). There was no significant difference between NWs and other groups. Lid wiper staining did not significantly differ between groups (P = .50) or correlate with the Nelson grade (Spearman r = 0.02, P = .08). Metaplasia of the LW epithelium was significantly greater in the early to moderate stages of CL. This supports the view that mechanical irritation is responsible for LW changes in CL wear. Ceasing CL wear seems to lead to recovery. Lid wiper staining did not reflect the underlying morphological changes.