Dunn's Multiple Comparison Test Research Articles

Plasma testosterone and arrhythmic events in male patients with arrhythmogenic right ventricular cardiomyopathy

AimsArrhythmogenic right ventricular cardiomyopathy (ARVC) is associated with life‐threatening ventricular arrhythmia and progressive ventricular dysfunction. Previous studies suggested that sex hormones play an important role in the onset and prognosis of ARVC. This study aimed to investigate the role of testosterone in predicting major adverse cardiac events in the Chinese ARVC cohort.Methods and resultsNinety‐nine ARVC patients (median age, 40 years; 70.7% male) and 96 healthy controls (median age, 41 years; 62.5% male) were enrolled. The circulating levels of testosterone were measured by enzyme‐linked immunosorbent assays (ELISA). The median follow‐up time of all ARVC male patients was 17 months (interquartile range/IQR 9–29). Cox proportional hazards regression was used to analyse the effect of plasma testosterone and other well‐described risk factors on malignant arrhythmic events in male ARVC patients. The male ARVC patients had significantly elevated levels of total testosterone [TT, 6.390 (4.438–8.768) ng/mL vs. 3.617 (2.073–4.479) ng/mL, P < 0.0001, data shown as the median with IQR], bioavailable testosterone [BT, 4.11 (1.990–6.545) ng/mL vs. 1.32 (0.7965–2.0350) ng/mL, P < 0.0001, median with IQR], and free testosterone [FT, 0.2055 (0.1000–0.4073) ng/mL vs. 0.0768 (0.0405–0.1105) ng/mL, P < 0.0001, median with IQR] than healthy male volunteer, whereas no differences were observed among female counterparts. There was no significant correlation between the baseline clinical characteristics and testosterone levels in male ARVC patients (Spearman's correlation test, P > 0.05). During the follow‐up, the levels of testosterone were higher in male patients who experienced malignant arrhythmic events (N = 22) than in those who did not (N = 25) [TT, 9.034 (7.222–15.370) ng/mL vs. 4.633 (3.363–6.375) ng/mL, P < 0.001; BT, 7.485 (2.070–9.163) ng/mL vs. 3.300 (1.685–4.690) ng/mL, P < 0.05; FT, 0.453 (0.221–0.758) ng/mL vs. 0.161 (0.075–0.337) ng/mL P < 0.05, data expressed as median (IQR) and adjusted by Dunn's multiple comparisons test], whereas such distinction was not observed among patients with significant structural progression events (N = 16). Through multivariable adjustments, the Cox regression analysis showed the level of plasma total testosterone (HR = 1.325, 95% confidence interval = 1.171–1.498, P < 0.001) was an independent predictor for malignant arrhythmic events.ConclusionsThe levels of plasma testosterone in ARVC male patients are higher than those in healthy males. Testosterone level, without relation to the baseline cardiac function and future significant structural progression events, is a strong predictor of future adverse arrhythmic events in male patients with ARVC. Therefore, our results suggest that testosterone may be a useful biomarker in arrhythmic risk prediction in the ARVC.

RAD54 promotes alternative lengthening of telomeres by mediating branch migration.

Cancer cells can activate the alternative lengthening of telomeres (ALT) pathway to promote replicative immortality. The ALT pathway promotes telomere elongation through a homologous recombination pathway known as break‐induced replication (BIR), which is often engaged to repair single‐ended double‐stranded breaks (DSBs). Single‐ended DSBs are resected to promote strand invasion and facilitate the formation of a local displacement loop (D‐loop), which can trigger DNA synthesis, and ultimately promote telomere elongation. However, the exact proteins involved in the maturation, migration, and resolution of D‐loops at ALT telomeres are unclear. In vitro, the DNA translocase RAD54 both binds D‐loops and promotes branch migration suggesting that RAD54 may function to promote ALT activity. Here, we demonstrate that RAD54 is enriched at ALT telomeres and promotes telomeric DNA synthesis through its ATPase‐dependent branch migration activity. Loss of RAD54 leads to the formation of unresolved recombination intermediates at telomeres that form ultra‐fine anaphase bridges in mitosis. These data demonstrate an important role for RAD54 in promoting ALT‐mediated telomere synthesis.

Comparison of cell salvage with one and two suction devices during cesarean section in patients with placenta previa and/or accrete: a randomized controlled trial.

BackgroundCell salvage has recently been recommended for obstetric use in cases with a high risk of massive hemorrhage during cesarean section (CS). However, limited data are available to support the use of one suction device to collect lost blood. This study aimed to investigate the volume of red blood cells (RBCs) salvaged and the components of amniotic fluid (AF) in blood salvaged by one suction device or two devices during CS in patients with placenta previa and/or accrete.MethodsThirty patients with placenta previa and/or accrete undergoing elective CS in the Women's Hospital of Zhejiang University School of Medicine were recruited for the present study from November 1, 2017 to December 1, 2018. The patients were randomly assigned to one of the two groups according to an Excel-generated random number sheet: Group 1 (n = 15), in which only one suction device was used to aspirate all blood and AF, and Group 2 (n = 15), in which a second suction device was mainly used to aspirate AF before the delivery of the placenta. Three samples of blood per patient (pre-wash, post-wash, and post-filtration) were collected to measure AF components. The salvaged RBC volumes were recorded. Continuous data of pre-wash, post-wash, and post-filtration samples were analyzed by using one-way analysis of variance with Tukey's test for multiple comparisons, or Kruskal-Wallis test with Dunn test for multiple comparisons. Comparisons of continuous data between Group 1 and Group 2 were conducted using Student's t test or Mann-Whitney U test.ResultsThe salvaged RBC volume was significantly higher in Group 1 than that in Group 2 (401.6 ± 77.2 mL vs. 330.1 ± 53.3 mL, t = 4.175, P < 0.001). In both groups, squamous cells, lamellar bodies, and fat were significantly reduced by washing (all P<0.001) and squamous cells were further reduced by filtering (P < 0.001). Squamous cells were found in six post-filtration samples (three from each group). Lamellar bodies and fat were completely removed by filtering. Insulin-like growth factor binding protein 1, alpha-fetoprotein, albumin, lactate dehydrogenase, and potassium were significantly reduced post-wash (all P < 0.05), with no further significant reduction after filtration in either group (all P > 0.05). The mean percentage of fetal RBCs post-filtration was (1.8 ± 0.8)% with a range of 1.0% to 3.5% and (1.9 ± 0.9)% with a range of 0.7% to 4.0% in Groups 1 and 2, respectively, showing no significant difference between the two groups (U = 188.5, P = 0.651).ConclusionCell salvage performed by one suction device could result in higher volume of salvaged RBCs and can be used safely for CS in patients with placenta previa and/or accrete when massive hemorrhage occurs.Trial registration numberChiCTR-INR-17012926, http://www.chictr.org.cn/ Chinese Clinical Trial Registry.

Influence of orthodontic loading on biomarkers levels around miniscrews

ObjectiveThe aim of this study was to evaluate the levels of Interleukin-1α (IL-1α), Interleukin-1β (IL-1β), Interleukin-1 receptor antagonist (IL-1Ra), Interleukin-10 (IL-10), Interleukin-13 (IL-13), Vascular endothelial growth factor (VEGF), Granulocyte-colony stimulating factor (G-CSF), and Growth related oncogene (GRO) in the peri-miniscrew implant crevicular fluid (MICF) under orthodontic loading. DesignThe study sample comprised 14 miniscrews immediately loaded and 17 unloaded ones. A load of 200gF was immediately applied to the miniscrews in the loaded group after the placement surgery. Peri-miniscrew implant crevicular fluid was collected at baseline, at day 7, and at day 21. The levels of the biomarkers were measured using a multiplexed bead immunoassay. Intergroup comparisons were made using Mann-Whitney test. Friedman and Dunn's multiple comparison tests were used to evaluate intragroup differences over time. ResultsAlthough no statistical differences were observed between the groups at any time point for any of the 8 biomarkers evaluated, there was a statistically significant increase (p < 0.02) in the levels of all the biomarkers over time on both groups. ConclusionsAn immediate loading of 200gF does not alter the balance in the inflammatory response in peri-miniscrew tissues.

A Transversal Study of Biochemical Profile, Urinalysis, UPC, Electrolytes and Blood Pressure in Dogs with Chronic Kidney Disease

Background: Chronic kidney disease (CKD) affects both dogs and cats, mainly elderly animals, due to tubulointerstitial inflammation associated with the increase of fibrosis through the excess deposition of extracellular matrix (ECM) which leads to decrease glomerular filtration. Many different underlying renal diseases can affect the kidneys of dogs such as congenital or acquired in origin. Therefore, the main objective of this transversal study was to evaluate the epidemiology through clinical and laboratory evaluation of 225 client-owned dogs with CKD.Materials, Methods &amp; Results: Complete blood count (CBC), urinalysis, and biochemical profile were retrospectively selected and evaluated from 225 client-owned dogs with CKD of both sexes, different ages, and breeds from the patient population of the Nephrology and Urology Small Animal Service of the Teaching Hospital of the School of Veterinary Medicine and Animal Science - São Paulo State University from 2011 to 2017. All dogs were divided in groups according to the International Renal Interest Society (IRIS) CKD grading and statistical analysis was performed according to Kruskal-Wallis non-parametric test complemented with Dunn's multiple comparisons test, and analysis of variance for the model with a factor complemented with the test of multiple comparisons of Tukey. In this retrospective study, we observed that most dogs in all groups were elderly (≥ 9 years old). CBC demonstrated lower RBC’s (P &lt; 0.005), hemoglobin (P &lt; 0.001), hematocrit (Ht%) [P &lt; 0.001] at the highest stage of the disease. However, urinary specific gravity (USG) did not demonstrate significant differences between the disease stages, but urinary protein: creatinine ratio (UPC) was statistically different (P &lt; 0.01) between IRIS-CKD stages 1 and 4. Furthermore, serum phosphate concentrations demonstrated significantly higher levels in dogs at IRI-CKD stage 4 compared with IRIS-CKD stage 3 (P &lt; 0.001).Discussion: The analysis of 225 dogs with CKD showed that 130 animals were elderly, older than 9 years, and according to previous studies, 15% of dogs over 10 years of age are diagnosed of CKD, presenting significant morbidity and mortality. Laboratory findings such as the presence of non-regenerative anemia is expected in dogs with CKD. In our study, the degree of anemia corresponded with the stage of the disease, similarly to serum creatinine concentrations. Another important laboratory finding in diagnosing CKD is the early detection of the kidney's abilities in concentrating its tubular filtrate. In this retrospective study, isosthenuria was not significant due to all dogs presented CKD as criteria of inclusion, especially those without azotemia, although, proteinuria was reported in 90% of the population investigated. Electrolyte imbalances are also expected in CKD. However, despite serum sodium, potassium, and total calcium did not demonstrate significant results, serum phosphate had its significance between IRIS-CKD stage 3 and stage 4. Hence, despite the age factor of most dogs in all groups, with the association of laboratory results such as serum creatinine, serum phosphorus, ionized calcium, erythrogram, isosthenuria, SBP, and the degree of proteinuria, it was possible to perform early diagnosis of CKD even in dogs with IRIS-CKD Stage 1 in a stable and hydrated patient. With the proper diagnostic, staging and substaging according to IRIS guidelines, these parameters can be monitored, predicting longevity and good quality of life, or progression of the disease with a more reserved prognosis.

Benefits and challenges in conducting long-term growth hormone therapy in patients with Prader-Willi syndrome

The purpose of this report is to comment the results from long-term growth hormone (GH) treatment of Bulgarian patients suffering from rare genetic disease-Prader-Willi syndrome (PWS) with reference to the age, body composition, complications and genetic etiology. Statistical analysis was performed by ANOVA with post hoc Kruskal-Wallis test and Dunn's multiple comparison tests. In 90% of the patients maternal uniparental disomy (mUPD) was found to be the cause of the disease. No cases due to imprinting defects are found. The BMI data shows no statistically significant difference between BMI at diagnosis (21.850), at the beginning of the GH therapy (21.852) and current BMI (24.09) - measured under the GH background. Early GH treatment allows to overcome arising obstacles in time and to improve the quality of life for PWS children and their families. During the fourteen years study period only ten patients were diagnosed with the disease. Ninety percent (n=9) of the children were found to be with maternal UPD (mUPD) and only one case was due to deletion in 15q11-13. These results are in agreement with other studies in the field which shows the need for reassessment and new robust statistical analysis of the frequency of genetic mechanisms for PWS.

Single-source dual-energy computed tomography for the detection of bone marrow lesions: impact of iterative reconstruction and algorithms.

To compare the diagnostic performance of different reconstruction algorithms of single-source dual-energy computed tomography (DECT) for the detection of bone marrow lesions (BML) in patients with vertebral compression fracture using MRI as the standard of reference. Seventeen patients with an age over 50 who underwent single-source DECT of the spine were included. The raw data (RD) were reconstructed using filtered back-projection (FBP) and iterative reconstruction (IR) with three iteration levels (IR1-IR3). Bone marrow images were generated using a three-material decomposition (3MD) and a two-material decomposition (2MD) algorithm and an RD-based approach. Three blinded readers scored the images for image quality and the presence of bone marrow lesions (BML). Only vertebrae with height loss were included. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were calculated. The different reconstructions were compared using Dunn's multiple comparison test. Thirty-nine vertebrae were included. IR(1-3) showed superior sensitivity (87.5%) compared to FBP (75%) using 3MD but was comparable to RD (83.3%). All 2MD images were inferior (sensitivity < 38%). The image quality score was significantly higher for 3MD-IR(1-3) compared to 3MD-FBP (p < 0.0001) and all 2MD data sets (p < 0.03). This pattern was also supported by the SNR and CNR measurements. RD showed no significant improvement compared to IR. The image quality of bone marrow images acquired with DECT can be improved by using IR compared with FBP. RD-based reconstruction does not offer significant improvement over image data-based reconstruction. 2MD algorithms are not suitable for BML detection.

27 Vitrification at the germinal vesicle stage does not trigger apoptosis in porcine oocytes and early embryos

Porcine immature oocytes can survive vitrification at high rates and retain their ability to undergo maturation and fertilization; however, the procedure reduces their competence for subsequent embryo development via unknown mechanisms (Somfai et al. 2014 Plos One 9, e97731). The aim of the present study was to clarify whether our vitrification procedure at the germinal vesicle stage triggers apoptosis in oocytes and subsequent developing embryos. Immature porcine cumulus-oocyte complexes obtained from slaughterhouse-derived ovaries were vitrified and warmed by our method (Appeltant et al. 2018 Cryobiology 85, 87-94) immediately after collection (vitrified group). The oocytes were equilibrated in 2% (vol/vol) ethylene glycol and 2% (vol/vol) propylene glycol for 13-15min. Then, they were vitrified by dropping them into liquid nitrogen in 2-μL microdrops of a medium composed of 17.5% ethylene glycol, 17.5% propylene glycol, 0.3M sucrose, and 50mgmL−1 polyvinylpyrrolidone. After warming, they were subjected to IVM, fertilization (IVF), and embryo culture using chemically defined media (Yoshioka et al. 2008 J. Reprod. Dev. 54, 208-213). From each collected batch, a group of oocytes was processed without vitrification (control group). Apoptosis was assayed in membrane-intact oocytes at the end of IVM and in cleavage-stage embryos on Day 2 after IVF (Day 0) by the CaspACE FITC-VAD-FMK In Situ Caspase Marker (Promega; Experiment 1), deoxynucleotidyl transferase (TdT) dUTP nick-end labelling (TUNEL; Experiment 2), and analysis of mRNA levels by RT-qPCR for the pro-apoptotic Bax and CASP3 genes (Experiment 3). Each experiment was replicated three times. Data were analysed by Kruskal-Wallis test followed by Dunn's multiple comparisons test. The mean survival rate of vitrified oocytes was 89.2%. There was no significant difference between the control and vitrified groups in relative caspase levels in IVM oocytes and in 2- to 4-cell embryos after IVF; however, significantly increased caspase activity (P&amp;lt;0.05) was detected in oocytes and embryos after treatment with 10 μM staurosporine (positive control). There was no significant difference between the control and vitrified groups in the proportion of TUNEL-positive oocytes (4.1 and 0.8%, respectively) and embryos (0 and 0%, respectively), whereas 96.6% of oocytes and 100% of cleavage stage embryos treated with 1000IUmL−1 deoxyribonuclease I (positive control) were proven to be TUNEL positive (P&amp;lt;0.05). Similar mRNA levels for Bax and CASP3 genes were detected in oocytes at the end of IVM and subsequent developing 4- to 8-cell embryos between the control and vitrified groups. In conclusion, vitrification of porcine oocytes at the germinal vesicle stage by our method did not trigger apoptosis in oocytes and subsequent developing embryos. This work was supported by the Japan Science and Technology Agency (JST)/Japan International Cooperation Agency (JICA) Science and Technology Research Partnership for Sustainable Development (SATREPS).

Diffusion Kurtosis Imaging and Intravoxel Incoherent Motion in Differentiating Nasal Malignancies.

To evaluate the usefulness of diffusion kurtosis imaging (DKI) and intravoxel incoherent motion (IVIM) in the differentiation of sinonasal malignant tumors (SNMTs) with different histological types. Retrospective observational and diagnostic study. Sixty-five patients with SNMTs who underwent DKI and IVIM were enrolled in this retrospective study, including 27 squamous cell carcinomas (SCCs), 13 olfactory neuroblastomas (ONBs), 14 malignant melanomas (MMs) and 11 lymphomas. The kurtosis (K) and diffusion coefficient (Dk) from DKI and the pure diffusion coefficient (D), pseudodiffusion coefficient (D*), perfusion fraction (f), and the product of D* and f (f∙D*) from IVIM were measured. Kruskal-Wallis and Dunn multiple comparison tests with Bonferroni correction, receiver operating characteristic curve, and logistic regression analyses were used for statistical analysis. Lymphomas demonstrated the highest K values but lowest Dk, D, D*, f, and f∙D* values among these four malignant tumors. ONBs exhibited high K values and MMs had highest D*, f, and f∙D* values. The cutoff value of ≤0.887 × 10-3 mm2 /sec for f∙D* provided a sensitivity, specificity, and an accuracy of 100%, 98.1%, and 98.5%, respectively, for differentiating lymphomas from the other three entities. The combination of f∙D* and D values showed a sensitivity of 92.9% and a specificity of 92.5% for the discrimination of MMs from ONBs and SCCs. The K value was useful for differentiating ONBs from SCCs, with a threshold value of 0.942 (sensitivity, 84.6%; specificity, 63.0%). The combined use of DKI and IVIM is helpful for differentiating among four histological types of SNMTs. 3 Laryngoscope, 2019.

Integrative Study of EZH2 Mutational Status, Copy Number, Protein Expression and H3K27 Trimethylation in AML/MDS Patients

Introduction Enhancer of Zeste Homolog 2 (EZH2), the catalytic domain of Polycomb Repressive Complex 2 (PRC2), mediates the repressive mark of trimethylation of histone H3 lysine 27 (H3K27me3). The EZH2 gene is located on chromosome (chr.) 7q36.1 (frequently deleted in AML/MDS). Its role in tumorigenesis appears to be context-dependent since both EZH2 overexpression and loss of function are associated with different types of cancer. In patients (pts) with MDS or MDS/MPN, loss-of-function EZH2 mutations (mut) are recurrently found, associated with a poor prognosis, and EZH2 is considered a tumor suppressor gene (TSG). In AML, the incidence of EZH2-mut is lower and less well-studied. We wished to determine the effects of EZH2-mut and copy number (CN) on EZH2 expression, and the consequences for H3K27me3 levels in vivo. Methods Fifty-eight pts (51 AML, 3 MDS, 4 MDS/MPN), median age 63.5 years (yr, range 20-86, almost all diagnosed between 2015 and 2017), with available sequencing data, EZH2 CN status and EZH2 expression data were studied. Mutation status was determined by next-generation sequencing (NGS) including a panel of 54 genes (Illumina Myeloid NGS panel). Metaphase cytogenetics and/or fluorescence in situ hybridization and single nucleotide polymorphism arrays (selected pts) were conducted to determine EZH2 CN status. Protein expression of EZH2 and H3K27me3 was assessed semi-quantitatively by immunohistochemistry (IHC) on formalin-fixed EDTA-decalcified paraffin-embedded bone marrow (BM) core biopsies. The Kruskal-Wallis test and Dunn's multiple comparison test were used to address whether EZH2 protein expression was associated with EZH2-mut and CN. Results The EZH2 gene showed mutations (mostly missense or nonsense, median variant allele frequency (VAF) 45%, range 7-63%) in 13/58 pts and was unmutated in 45/58 pts. EZH2-mut pts had a median of 4 mutations. Additional mutations were most frequently found in ASXL1 (10/13; median VAF 35%, range 19-48%), less frequently in TET2, RUNX1, STAG2 (3/13 each), NRAS (2/13), and DNMT3A (1/13). In contrast, EZH2-wt pts had a median of 2 mutations, most frequently in DNMT3A (12/45), followed by NRAS (10/45), IDH1 (9/45), FLT3 (8/45), and NPM1 (7/45). Regarding chr. 7, 43 pts had no detectable deletion, 15 had 7q-/-7. Notably, the incidence of EZH2-mut was similar in pts with 7q-/-7 lesions (3/15, i.e. 20%) and pts with normal chr. 7 (10/43, 23%). EZH2 expression in neoplastic BM cells ranked from no (score 0) to strong expression (score 3). While the hematopoietic BM cells of healthy donors usually showed a moderate EZH2 expression (score 2), in our cohort the score was 0 in 3, 1 in 13, 2 in 23, and 3 in 19 pts, respectively. We next asked whether EZH2 protein expression differed between pts depending on EZH2-mut and chr. 7 status. In Figure 1, 4 subgroups are depicted, showing highest expression in pts with EZH2-wt and either no chr. 7 abnormalities (group A, n=33) or 7q-/-7 (group C, n=12). In comparison, expression was significantly reduced in pts with EZH2-mut and no chr. 7 abnormalities (group B, n=10), and lowest in pts with EZH2-mut and chr. 7 abnormalities (group D, n=3), p&amp;lt;0.05 (indicated by asterisks). Since functional EZH2 protein is necessary for the trimethylation of H3K27, the presence of this histone mark was also determined semi-quantitatively by IHC in 40 pts. H3K27me3 levels were variable, and a test for a possible association between EZH2 and H3K27me3 levels by linear regression analysis did not show an association (R²=0.13). Sixty-two % of EZH2-mut (median age 71 yr) and 51% of EZH2-wt pts (median age 61 yr) received allografting. Median overall survival in EZH2-mut pts was 16.1 months compared to 23.5 in EZH2-wt pts (p=0.16). Conclusions Inactivating EZH2 mutations are infrequent events in AML. In our study, they were strongly associated with mutations of ASXL1 (also involved in PRC2 function). We did not observe overrepresentation of EZH2-mut in 7q-/-7 pts, in line with Bejar et al. (N Engl J Med 2011); this is in contrast to the frequent cooperative events between TSG mutations and deletions, e.g. of TP53. Functionally, EZH2 mutations appeared to have a stronger effect on decreased EZH2 expression than hemizygosity. Global H3K27me3 levels were not altered by EZH2 reduction, which could be due to compensatory upregulation of EZH1, supporting the clinical development of EZH1/2 inhibition. Disclosures No relevant conflicts of interest to declare.

Narcotic Use in Multiple Myeloma Patients at the Time of ASCT and One-Year Follow up

Background: Bone pain is one of the most common presentations of multiple myeloma and nearly all patients have skeletal involvement in the course of disease. Consequently, many patients require narcotics for symptom management at the time of diagnosis but the long term impact of MM treatment on pain control remains uncertain. With the advent of combination therapy for MM with novel agents followed by transplant and then maintenance therapy, clinical response is nearly universal and greater than 30% of patients achieve a serologic complete response. Therefore, we examined the impact of modern myeloma-directed therapy and high response rates on the use of narcotics up to 1 year after transplant in this group of patients. Methods: A retrospective review of data collected from the Rutgers-CINJ database was conducted. All patients who received induction inducing therapy (e.g. bortezomib, lenalidomide and dexamethasone or cyclophosphamide, bortezomib and dexamethasone) followed by high dose melphalan and autologous stem cell transplant (ASCT) and who had adequate post-transplant follow up (at least 100 days) were included. Morphine use was assessed at the time of transplant and at follow-up visits. All opiates (e.g. oxycodone, fentanyl, MS contin, etc.) where converted to morphine equivalents/day (ME/day) and recorded. Treatment responses were determined based on the International Myeloma Working Group Response Criteria. We compared the incidence and amount of narcotic use over time using one-way analysis of variance (ANOVA) and Dunn's multiple-comparison test. Results: 189 patients were included in the analysis. 38% were using opiates at the time of transplant. At 100 days post-transplant 35.5% were using opiates and at 1 year post transplant 30.9% were using opiates (p=0.04) . Average opiate use was 74.1 ME/day (95% CI: 55.2 to 92.9), 69.45 ME/day (95% CI: 50.5 to 88.3), and 70.78 ME/day (95% CI: 43.5 to 98) for each of the aforementioned time points (p=0.088) (Figure 1). For example, 74 ME/day would be equal to approximately 50 mg of Oxycodone daily. 74 patients were active opiate users at the time of transplant (Table 1). Response to myeloma treatment (remission, progression, relapse) was not different in opiate-using patients at the time of transplant or at 100 days and 1 year after transplant (Table 2). Conclusion : Early studies from the 1960's reported on the rapid reduction of bone pain in treatment responsive patients with MM (Hogstrata et al.). Recently, treatment modalities for MM have significantly improved, leading to markedly increased remission rates (&gt;90%), progression-free and overall survival. With dramatically increased serologic responses, one would hypothesize that there would be a decline in pain and a subsequent decrease in opiate use. However, in this retrospective chart review we found that 30% of multiple myeloma patients continued to use opiates following transplant and that ongoing use of opiates appeared to be independent of clinical response. Interestingly, at the time of transplant active opiate users had a CR of 27.2% while the non-opiate using patients had a CR of 13.3%. Although there was a slight decline in the number of opiate users at the 1 year follow up, 33% of opiate users continued at their original level of opiate use and 22% increased their opiate use at 1 year post ASCT. Persistent long term use of opiates is of particular concern given that the survival of patients with MM has significantly improved with the use of novel agents and autologous transplant with an increasing number of patients surviving 10 years or more. In patients who have achieved an excellent response (e.g. serologic CR and negative PET scan), ongoing narcotic use should be addressed at each visit and other advanced pain management techniques should be considered. Disclosures No relevant conflicts of interest to declare.

Evaluation of the host cytokine response in dogs with sepsis and noninfectious systemic inflammatory response syndrome.

To quantify plasma cytokine concentrations in dogs with sepsis and noninfectious systemic inflammation and to evaluate the association between plasma cytokines and outcome in dogs with sepsis. Prospective, observational cohort study. University teaching hospital. Forty-five dogs with sepsis, 10 dogs with noninfectious systemic inflammation (nSIRS), and 15 healthy controls were consecutively enrolled from June 2015 to February 2016 and followed to hospital discharge. Dogs with sepsis satisfied ≥2 SIRS criteria and had a documented or highly suspected bacterial infection. Dogs with nSIRS satisfied ≥2 SIRS criteria but had no evidence of infection. Dogs<3kg and those with documented coagulopathy were excluded. Measurement of inflammatory cytokines and high-mobility group box-1 (HMGB-1) was performed on each group. High-mobility group box-1 concentrations were analyzed by ELISA. Plasma concentrations of 13 cytokines were measured in singlet using multiplex magnetic bead assays. Kruskal-Wallis with Dunn's multiple comparison tests were used to compare biomarker concentrations between groups. Mann-Whitney U-tests were used to compare biomarker concentrations between survivors and nonsurvivors. Associations between biomarkers were evaluated using Spearman's correlation coefficients. Independent outcome predictors were identified using multivariable logistic regression. Alpha was set at 0.05. Concentrations of interleukin (IL)-6, C-X-C motif chemokine (CXCL)-8, keratinocyte-derived chemokine (KC)-like, C-C motif chemokine ligand 2 (CCL2), and HMGB-1 were significantly greater in dogs with sepsis versus healthy controls (all P≤0.034). In dogs with sepsis, only CCL2 was independently associated with survival (odds ratio [OR] 0.996, 95% CI 0.993-0.999, P=0.004). A cut-off of 385 pg/mL for CCL2 was 80% sensitive and 91.4% specific for nonsurvival (area under the ROC curve [AUROC] 0.866). Dogs with sepsis have significantly increased concentrations of HMGB-1 and inflammatory cytokines, including IL-6, CXCL8, and KC-like. Increased CCL2 concentration is a negative prognostic indicator in dogs with sepsis. These findings should be confirmed using duplicate analyses in larger, distinct populations of dogs with sepsis before applying them to clinical patients.

Apical Transportation and Centering Ability After Root Canal Filling Removal Using Reciprocating and Continuous Rotary Systems: A CBCT Study.

Objective To evaluate the apical transportation and centering ability promoted by reciprocating and continuous rotary systems after root canal filling removal. Materials and Methods After obturation, 40 mesial root canals of mandibular molars were distributed into four groups ( n = 20) for filling material removal: PTU group–F2 instrument (25.08) of ProTaper Universal system; R25 group–R25 instrument (25.08) of Reciproc system; X2 group–X2 instrument (25.06) of ProTaper Next system and X3 group–X2 instrument (25.06) of ProTaper Next system, followed by X3 instrument (30.07). Cone-beam computed tomographic analysis was performed before and after filling material removal for acquisition of apical images. Apical transportation (AT) and its direction, and centering ability (CA), were assessed using the equations AT = (X1–X2)–(Y1–Y2) and CA = (X1–X2/Y1–Y2 or Y1–Y2/X1–X2), respectively. Data were submitted to the nonparametric Kruskal–Wallis and Dunn multiple comparison tests ( p < 0.05) for statistical analysis. Results There was no statistically significant difference among groups for AT ( p > 0.05), with a tendency toward transportation in the distal direction. Also, there was no statistically significant difference among groups regarding CA ( p > 0.05). Conclusions The different systems, including ProTaper Next, caused AT within the acceptable clinical limit after filling removal. In addition, none of the tested systems presented adequate CA.

Antimicrobial efficacy of cordless sonic or ultrasonic devices on Enterococcus faecalis-infected root canals.

The aim of the present study was to evaluate the immediate and residual antimicrobial activity of cordless sonic or ultrasonic devices on Enterococcus faecalis (Efaecalis)-infected teeth. A total of 140 single-rooted extracted teeth with Efaecalis were grouped as follows (N=15): conventional syringe irrigation with 3% sodium hypochlorite activated by ultrasonic device (group 1) or cordless ultrasonic device (group 2), EndoActivator (group 3) or without activation (group 4), conventional syringe irrigation with sterile bi-distilled water and ultrasonic device (group 5), EndoUltra (group 6), EndoActivator (group 7), or without activation (group 8). The remaining 20 teeth were used for positive and negative controls. Colony-forming units (CFU) and turbidity were recorded from bacteriological samples taken before and after irrigation and after 24hours of re-incubation. Data were statistically analyzed using Student's t test, Mann-Whitney test, Kruskal-Wallis test, and Dunn's multiple comparison tests (P<.05). Groups 1 and 2 showed higher antimicrobial efficacy than groups 3 and 4 (P<.05). No bacteria reduction was found in groups 5-8 (P>.05). After 24hours, regrowth of bacteria was not significantly different for all groups (P>.05). Ultrasound produced lower CFU and turbidity after treatment and after re-incubation of 24hours than sonic or no activation.

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.