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Background: Mountain communities have unique issues, being generally geographically remote, and politically and socially neglected. The objective was to evaluate the health and healthcare access of children in Tissar, a mountain village in Gilgit-Baltistan. Subjects and methods: This was a cross-sectional study conducted in August 2021, in Tissar, Gilgit-Baltistan (GB), in Northern Pakistan. Interviews were conducted of adults over the age of 18 years, who had at least one child of age12 years and below, using a convenience sampling method. Descriptive summary statistics were computed using statistical software R version 4.1.2. Poisson regression was used to find potential risk factors associated with diarrhea and pneumonia. Results: Semi-structured interviews were conducted on 400 participants. Although 363 (90.8%) people grew their own food; 203 (50.7%) still needed to purchase food with only 213 (53.2%) receiving up to three meals a day. Access to healthcare was perceived as inadequate, with only 42 (10.5%) satisfied with the healthcare their children were receiving. Up to 268 (67%) needed to borrow money or could not afford healthcare. Annual occurrence of diarrhea and pneumonia was reported in 100% households, most reporting repeated episodes in one year per child. On Poisson Regression, number of meals per day was negatively associated with diarrhea (rate ratio=0.646, 95%CI=0.591-0.706, p6 versus

Patients with bladder cancer who undergo intestinal urinary diversion may be at increased risk for bone fractures thought to be secondary to chronic metabolic acidosis and ensuing bone loss. Our main objective was to assess whether patients who undergo intestinal urinary diversion are at increased risk for fracture.Patients who underwent intestinal urinary diversion between 1994 and 2014 in Ontario, Canada were identified using linked administrative databases. Patients were categorized as undergoing diversion for bladder cancer or nonbladder cancer causes and matched 4:1 to a healthy cohort. We determined incidence rates of the incidence of fractures per 100 person-years. Multivariable Cox proportional hazards models were used to evaluate the impact of intestinal urinary diversion on the risk of fracture.Overall 4,301 patients with and 907 without bladder cancer underwent intestinal urinary diversion. The fracture incidence rate was significantly greater in the bladder cancer and nonbladder cancer cohorts compared to respective matched controls. In the bladder cancer cohort vs matched controls there were 4.41 vs 2.63 fractures per 100 person-years and in the nonbladder cancer cohort vs matched controls there were 5.67 vs 3.51 fractures per 100 person-years (each p0.001). On multivariable analysis patients who underwent intestinal urinary diversion for bladder cancer or nonbladder cancer reasons had significantly shorter fracture-free survival compared to the respective matched cohorts (HR 1.48, IQR 1.35-1.63, and HR 1.48, IQR 1.31-1.69, respectively).Our results demonstrated that regardless of age patients with intestinal urinary diversion are at increased risk for bone fractures compared to the general population. Our findings are in line with previous reports and support the need for bone health monitoring.

Highly sensitive and specific urinary biomarkers for the early detection of bladder cancer (BC) to improve the performance of urinary cytology are needed.To investigate the usefulness of methylation markers in voided urine to identify BC presence and grade.Using genome-wide methylation strategies in Toronto, Canada and Liège, Belgium, we have identified differentially methylated genes (TWIST1, RUNX3, GATA4, NID2, and FOXE1) in low-grade vs. high-grade BC tissue and urine. We accrued urine samples from 313 patients using a 2:1 ratio in a case-control setting from Toronto, Canada, Halifax, Canada, and Zurich, Switzerland. We studied the usefulness of these 5 methylated genes to identify BC and discriminate cancer grade in voided urine specimens. Urinary cell sediment DNA was evaluated using qPCR-based MethyLight assay. Multivariable logistic regression prediction models were created.We included 211 BC patients (180 nonmuscle invasive) and 102 controls. In univariate analyses, all methylated genes significantly predicted BC vs. no BC, and high grade vs. low grade (all P0.05). In multivariable analysis, NID2, TWIST1, and age were independent predictors of BC (all P0.05). Sensitivity of NID2 and TWIST1 to predict BC and BC grade was 76.2% and 77.6%, respectively, whereas specificity was 83.3% and 61.1%, respectively. Multivariable models predicting BC overall and discriminating between high-grade and low-grade BC reached area under the receiver operating characteristics curves of 0.89 and 0.78, respectively.This multi-centric study in a real life scenario (different countries, techniques, and pathologists) supports the promise of epigenetic urinary markers in noninvasively detecting BC. With sensitivities and specificities in the range of 80%, the overall performance characteristics of this panel of methylated genes probably does not allow such signature to significantly alter clinical care at this stage but is worth further studying for instance in BC surveillance or screening in high-risk populations.

Introduction: Non-muscle-invasive bladder cancer (NMIBC) accounts for 75‒85% of all urothelial bladder cancers (UBC). Many UBC patients are also afflicted by diabetes mellitus (DM). It has been postulated that several oral hypoglycemic agents could impact disease-specific survival (DSS), but the data are sparse among NMIBC patients. Our primary objective was to evaluate the impact of metformin on DSS and overall survival (OS) in NMIBC patients.Methods: This is a retrospective, population-based study that used linked administrative databases to identify diabetic patients ≥66 years who were subsequently diagnosed with NMIBC in Ontario between 1992 and 2012. Cumulative use of metformin and other hypoglycemic agent were calculated before and after NMIBC diagnosis. DSS and OS were estimated using multivariable competing risk and Cox proportional hazards models, respectively.Results: A total of 1742 subjects were included in the study. After a median followup of 5.2 years, 1122 (64%) had died, including 247 (15%) deaths as a result of UBC. On multivariable analysis, cumulative duration of metformin use after NMIBC diagnosis did not appear to impact DSS (hazard ratio [HR] 1.1; 95% confidence interval [CI] 0.92‒1.2), whereas glyburide use appeared to have a detrimental effect (HR 1.17; 95% CI 1.02‒1.3). None of the other hypoglycemic agents had an impact on OS.Conclusions: In this large, population-based study, we have provided further evidence that metformin use does not significantly impact DSS among diabetic patients diagnosed with NMIBC. However, our findings demonstrate that glyburide use inversely affects DSS. The detrimental effect of glyburide on DSS will require further validation.

Mediastinal radiation therapy (RT) for Hodgkin lymphoma (HL) is associated with late cardiotoxicity, but there are limited data to indicate which dosimetric parameters are most valuable for predicting this risk. This study investigated which whole heart dosimetric measurements provide the most information regarding late cardiotoxicity, and whether coronary artery dosimetry was more predictive of this outcome than whole heart dosimetry.A random sample of 125 HL patients treated with mediastinal RT was selected, and 3-dimensional cardiac dose-volume data were generated from historical plans using validated methods. Cardiac events were determined by linking patients to population-based datasets of inpatient and same-day hospitalizations and same-day procedures. Variables collected for the whole heart and 3 coronary arteries included the following: Dmean, Dmax, Dmin, dose homogeneity, V5, V10, V20, and V30. Multivariable competing risk regression models were generated for the whole heart and coronary arteries.There were 44 cardiac events documented, of which 70% were ischemic. The best multivariable model included the following covariates: whole heart Dmean (hazard ratio [HR] 1.09, P=.0083), dose homogeneity (HR 0.94, P=.0034), male sex (HR 2.31, P=.014), and age (HR 1.03, P=.0049). When any adverse cardiac event was the outcome, models using coronary artery variables did not perform better than models using whole heart variables. However, in a subanalysis of ischemic cardiac events only, the model using coronary artery variables was superior to the whole heart model and included the following covariates: age (HR 1.05, P.001), volume of left anterior descending artery receiving 5 Gy (HR 0.98, P=.003), and volume of left circumflex artery receiving 20 Gy (HR 1.03, P.001).In addition to higher mean heart dose, increasing inhomogeneity in cardiac dose was associated with a greater risk of late cardiac effects. When all types of cardiotoxicity were evaluated, the whole heart variable model outperformed the coronary artery models. However, when events were limited to ischemic cardiotoxicity, the coronary artery-based model was superior.

Background Non–muscle-invasive bladder cancer (NMIBC) is especially prevalent among the elderly. Many patients with NMIBC also have significant concomitant comorbidities, including cardiovascular diseases and hypercholesterolemia. Statins are the most commonly used cholesterol-depleting agents, and they may possess anticancer properties. The objective of this population-based study was to evaluate the effect of statins on the survival of individuals diagnosed with NMIBC. Methods This is a retrospective population-based cohort study that used administrative databases to identify individuals 66 years of age and older who were diagnosed with NMIBC between 1992 and 2012. Subjects with documented use of statins before they were 66 years of age were excluded from the analysis. Cumulative daily use of statins was calculated before and after the diagnosis of NMIBC. Their effect on cancer-specific survival and overall survival was estimated using a multivariable competing risk and Cox proportional hazards model, respectively. Results The final cohort was composed of 13,811 individuals≥66 years diagnosed with NMIBC. Of these, 4,748 individuals (34%) were exposed to statins during follow-up. The median statin exposure after NMIBC diagnosis was 21.4 months (interquartile range: 7.8–45.4). After a median follow-up of 7.1 years (interquartile range: 4.0–11.3) from NMIBC diagnosis, 8,900 (64%) individuals had died. The cumulative use of statins after NMIBC diagnosis did not significantly affect cancer-specific survival ( P = 0.10). However, its cumulative use after NMIBC diagnosis was associated with a better overall survival ([0.93; 95% CI: 0.91–0.96], per year of use). Conclusions This large population-based study has provided evidence that cumulative statin use was not associated with an improved cancer-specific survival among individuals with NMIBC. However, our findings did demonstrate that statin users had a better overall survival than nonusers.

INTRODUCTION AND OBJECTIVES:Aberrant DNA methylation is very common in bladder cancer (BC) pathogenesis and progression. These epigenetic alterations are detectable in voided urine and might be use...

Background Cardiac surgery patients are at high risk for postoperative bleeding. Intravenous (IV) tranexamic acid (TxA) is a commonly used antifibrinolytic drug, but is associated with postoperative seizures. We conducted this pilot randomized controlled trial (RCT) to determine the feasibility of a larger trial that will be designed to investigate the impact of TxA administration route, intrapericardial (IP) vs IV, on postoperative bleeding and seizures. Methods In this single-center, double-blinded, pilot RCT we enrolled adult patients undergoing nonemergent on-pump cardiac operations through a median sternotomy. Participants were randomized to IP or IV TxA groups. The primary outcomes were cumulative chest tube drainage, transfusion requirements, and incidence of postoperative seizures. Results A total of 97 participants were randomized to the intervention and control groups. Baseline characteristics were similar in both groups. Most participants underwent a CABG and/or aortic valve replacement. There was no statistical difference. The IP TxA group was found to have a tendency for less chest tube drainage in comparison to the IV TxA group, 500.5 (370.0-700.0) and 540.0 (420.0-700.0) mL, respectively, which was not statistically significant (P = 0.2854). Fewer participants in the IP TxA group with cardiac tamponade and/or required a reoperation for bleeding and fewer packed red blood cell transfusions. None of the IP TxA group developed seizure vs one from the IV TxA group. Conclusion This is the first known pilot RCT to investigate the role of TxA route of administration in open cardiac surgery. Intrapericardial TxA shows promising results with decreased bleeding, transfusion requirements, reoperations, and postoperative seizures. A larger RCT is needed to confirm these results and lead to a change in practice.

Age-related macular degeneration (AMD) is a disease of the elderly in which central vision is lost because of degenerative changes of the macula. The current study investigated the association of single-nucleotide polymorphisms (SNPs) with AMD in the Pakistani population. Four SNPs were analyzed in this study: rs1061170 in the CFH, rs429608 near CFB, rs2230199 in the C3, and rs10490924 in ARMS2/HTRA1. This case-control association study was conducted on 300 AMD patients (125 wet AMD and 175 dry AMD) and 200 unaffected age- and gender-matched control individuals. The association of the SNP genotypes and allele frequency distributions were compared between patients and healthy controls, keeping age, gender, and smoking status as covariates. A significant genotype and variant allele association was found of rs10490924 in ARMS2/HTRA1 with wet AMD, while the SNPs in CFH, CFB, and C3 were not associated with AMD in the current Pakistani cohort. The lack of association of CFH, CFB, and C3 may be attributed to limited sample size. This study demonstrates that genetic causative factors of AMD differ among populations and supports the need for genetic association studies among cohorts from various populations to increase our global understanding of the disease pathogenesis.

Signaling via the pre-T cell antigen receptor (pre-TCR) and the receptor Notch1 induces transient self-renewal (β-selection) of TCRβ(+) CD4(-)CD8(-) double-negative stage 3 (DN3) and DN4 progenitor cells that differentiate into CD4(+)CD8(+) double-positive (DP) thymocytes, which then rearrange the locus encoding the TCR α-chain (Tcra). Interleukin 7 (IL-7) promotes the survival of TCRβ(-) DN thymocytes by inducing expression of the pro-survival molecule Bcl-2, but the functions of IL-7 during β-selection have remained unclear. Here we found that IL-7 signaled TCRβ(+) DN3 and DN4 thymocytes to upregulate genes encoding molecules involved in cell growth and repressed the gene encoding the transcriptional repressor Bcl-6. Accordingly, IL-7-deficient DN4 cells lacked trophic receptors and did not proliferate but rearranged Tcra prematurely and differentiated rapidly. Deletion of Bcl6 partially restored the self-renewal of DN4 cells in the absence of IL-7, but overexpression of BCL2 did not. Thus, IL-7 critically acts cooperatively with signaling via the pre-TCR and Notch1 to coordinate proliferation, differentiation and Tcra recombination during β-selection.

Purpose: Three genes VSX1, HGF and LOX have been previously reported to be causative of keratoconus (KC) in different populations. In the current study we investigated the role of these genes in three autosomal recessive Pakistani families and 100 sporadic KC cases from Pakistan. Methods: Mutation analysis of VSX1 gene was done by Sanger sequencing of three KC families, while 100 sporadic cases including 10 cases with positive maternal family history were sequenced for mutation hotspot exon 2 of the gene. For the sporadic cases, case-control association analysis was also performed for KC associated reported polymorphisms in HGF (rs17501108 and rs3735520), and LOX (rs1800449, rs2288393 and rs2956540) gene through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: Screening of the VSX1 did not reveal any mutation in KC panel, a synonymous polymorphism rs12480307 (c.546A>G; p.Ala182Ala) in exon 3 was observed in one of the proband who inherited it homozyogusly from the carrier parent. This polymorphism was not observed in sporadic KC cases. In addition no significant association of the HGF and LOX SNPs were observed through case-control association analysis of sporadic cases. Conclusion: This is the first report of KC genetics from Pakistan where we excluded the role of VSX1 in Pakistani KC panel, in addition none of the previously reported SNPs of HGF and LOX have any involvement in disease pathogenesis in Pakistani KC cases.

Robotic surgery has emerged as a minimally invasive alternative to traditional laparoscopy. Robotic surgery addresses many of the technical and ergonomic limitations of laparoscopic surgery, but the literature regarding clinical outcomes in colorectal surgery is limited. We sought to compare robotic and laparoscopic colorectal resections with respect to 30-day perioperative outcomes.The American College of Surgeons National Surgical Quality Improvement Program database was used to identify all patients who underwent robotic or laparoscopic colorectal surgery in 2013. We performed a logistic regression analysis to compare intraoperative variables and 30-day outcomes.There were 8392 patients who underwent laparoscopic colorectal surgery and 472 patients who underwent robotic colorectal surgery. The robotic cohort had a lower incidence of unplanned intraoperative conversion (9.5% v. 13.7%, p = 0.008). There were no significant differences between robotic and laparoscopic surgery with respect to other intraoperative and postoperative outcomes, such as operative duration, length of stay, postoperative ileus, anastomotic leak, venous thromboembolism, wound infection, cardiac complications and pulmonary complications. On multivariable analysis, robotic surgery was protective for unplanned conversion, while male sex, malignancy, Crohn disease and diverticular disease were all associated with open conversion.Robotic colorectal surgery has comparable 30-day perioperative morbidity to laparoscopic surgery and may decrease the rate of intraoperative conversion in select patients.La chirurgie robotique est de plus en plus utilisée comme option de rechange peu effractive à la laparoscopie classique. La robotique permet de remédier à bon nombre des restrictions techniques et ergonomiques de la chirurgie laparoscopique, mais peu d'articles font état des résultats cliniques en chirurgie colorectale. Nous avons donc cherché à comparer les 2 techniques de résection colorectale en ce qui concerne les résultats peropératoires dans les 30 jours suivant l'intervention.À l'aide de base de données du National Surgical Quality Improvement Program de l'American College of Surgeons, nous avons recensé tous les patients ayant subi une résection colorectale par chirurgie laparoscopique ou robotique en 2013. Nous avons ensuite mené une analyse de régression logistique pour comparer des variables peropératoires et les résultats après 30 jours.En tout, 8392 patients avaient subi une chirurgie colorectale par laparoscopie pendant la période visée, et 472 avaient subi une intervention par chirurgie robotique. Le second groupe avait une incidence plus faible de conversion peropératoire imprévue (9,5 % par rapport à 13,7 %;Les taux de morbidité peropératoire après 30 jours pour une résection colorectale par chirurgie robotique et une intervention par chirurgie laparoscopique sont comparables. La chirurgie robotique pourrait de plus réduire le taux de conversion peropératoire chez certains patients.

This article proposes a robust model for parameter estimation when modeling in the presence of latent heterogeneity of population. Mixture of regression models are used for modeling when the data are a mixture of subgroups to allow for heterogeneity of the population. Parameter estimates from standard mixture of linear regression models are sensitive to atypical observations. To study mixtures of linear regression models, we introduce a class of robust estimators, called S-estimators. We investigate their breakdown point in mixture of linear regression models. It is expected that the robust S-estimators can achieve the high breakdown point in the contaminated data from the heterogenous populations. This model presents a unified, robust framework and parameter estimation is achieved via an expectation-conditional maximization (ECM) algorithm. This new family of robust mixture models is validated through Monte Carlo simulations. The application to real life data sets has shown that use of robust S-estimator...

This article addresses the use of different biweight functions in high breakdown mixture discriminant analysis approach of Bashir and Carter (2005). The translated biweight S function of Rocke (1996) is used in the robust estimation of the mixture distributions parameters. As in discriminant analysis, the main purpose is to classify the test observations with greater accuracy, so the estimators producing smaller errors of misclassification are preferred. In the simulation studies, the use of Tukey's biweight function was proved to be more effective in producing better classification results as compared to the translated biweight function.

Robust S-estimation is proposed for multivariate Gaussian mixture models generalizing the work of Hastie and Tibshirani (J. Roy. Statist. Soc. Ser. B 58 (1996) 155). In the case of Gaussian Mixture models, the unknown location and scale parameters are estimated by the EM algorithm. In the presence of outliers, the maximum likelihood estimators of the unknown parameters are affected, resulting in the misclassification of the observations. The robust S-estimators of the unknown parameters replace the non-robust estimators from M-step of the EM algorithm. The results were compared with the standard mixture discriminant analysis approach using the probability of misclassification criterion. This comparison showed a slight reduction in the average probability of misclassification using robust S-estimators as compared to the standard maximum likelihood estimators.

In the case of a large number of feature vector variables, using multivariate Gaussian mixture models, discrimination in a reduced subspace is studied, generalizing Hastie and Tibshirani's (1996) work, to a situation in which the outliers are present in the data. In the case of the Gaussian Mixture models, the reduced rank discriminant analysis is equivalent to the weighted rank k linear discriminant analysis (LDA). The reduced rank solution in the mixtures of multivariate Gaussian models was obtained from the full rank robust mixture solution. The classification in the new dimensions was compared with the discriminant analysis approach based on the original coordinates, using robust S-estimators. In most of the cases, the robust reduced rank mixture discriminant analysis (mda) performed better for the test data. However, for the case of common component covariance being diagonal, the robust reduced rank mixture discriminant analysis performed better than the robust full rank mixture discriminant...

e16003 Background: Multidisciplinary management improves complex treatment decision making in cancer care, but its impact for bladder cancer (BC) has not been documented. While radical cystectomy (RC) is currently viewed as the standard of care for muscle-invasive bladder cancer (MIBC), radiotherapy-based, bladder-sparing trimodal therapy (TMT) combining transurethral resection of bladder tumor, chemotherapy for radiation sensitization and external beam radiotherapy has emerged as a valid treatment option. In the absence of randomized studies, we compared the oncological outcomes between patients managed by RC or TMT using a propensity-score matched cohort analysis. Methods: Patients seen in our multidisciplinary bladder cancer clinic (MDBCC) from 2008 to 2013 were retrospectively reviewed. Those who received TMT for MIBC were identified and matched (for gender, cT and cN stage, ECOG status, Charlson comorbidity score, treatment date, age, CIS, hydronephrosis) using propensity scores, to patients who underwent RC. Overall survival and disease-specific survival (DSS) were assessed with Cox Proportional hazards modeling and competing risk analysis, respectively. Results: 112 patients with MIBC were included after matching, 56 treated with TMT and 56 by RC. Median age was 68.0 years and 29.5% were cT3/cT4. At a median follow up of 4.51 years, there were 20 (35.7%) deaths (13 from BC) in the RC group and 22 (39.3%) deaths (13 from BC) in the TMT group. 5 year DSS was 73.2% and 76.6%, in the RC and TMT groups, respectively (p = 0.49). Salvage cystectomy was performed in 6/56 TMT patients (10.7%). Conclusions: In the setting of a MDBCC, TMT yielded survival outcomes similar to matched RC patients. Appropriately selected MIBC patients should be offered the opportunity to discuss various treatment options including organ-sparing TMT.

Background Liver inflammation due to HCV infection leads to fibrosis, which is an independent predictor of treatment response to interferon therapy in Chronic Hepatitis C (CHC) patients. This relationship has not been studied for liver inflammation on pretreatment liver biopsy and End of Treatment Response (ETR). ALT is a less invasive test than liver biopsy for measuring liver inflammation. Aim of this study was to compare ETR to Interferon α (recombinant Interferon) & Ribavirin in CHC patients having higher and lower grades of liver inflammation and to determine the diagnostic accuracy of pretreatment ALT for grades of liver inflammation. Methods A retrospective cohort of 876 naïve CHC patients, who completed Interferon α & Ribavirin for 24 weeks, was studied for ETR. Pretreatment grade of inflammation on liver biopsy was taken as the exposure variable. It was classified as high if there was moderate or severe and low if there was minimal or mild. Multivariable logistic regression modeling was performed. Diagnostic accuracy of pretreatment ALT for liver inflammation grades was determined by computing Area Under the Receiver Operator Curve (AUROC). Results Of all patients, 672 having diagnostic liver biopsy and ETR available were analyzed. Among them, 103 had high and 569 had low grades of liver inflammation. Mean age was 36.9 (SD 9.1) years, with patients with high grades being older than those with low grades inflammation (p =���0.03). High grades of liver inflammation was associated with ETR (RR 1.17, 95% CI 1.12–1.18) adjusting for age, Total Leukocyte count (TLC) and pretreatment levels of ALT, irrespective of liver fibrosis. This relation remained significant for ‘bridging fibrosis and cirrhosis’ and not for ‘no’ or ‘portal fibrosis’. AUROC of pretreatment ALT for males and females was moderately accurate for severe inflammation compared to minimal inflammation and less accurate for high grades compared to low grades. Conclusions ETR in patients with higher grades of liver inflammation was 17% higher than those with lower grades irrespective of fibrosis and 9% higher for bridging fibrosis and cirrhosis. Pretreatment ALT was moderately accurate for severe inflammation only on liver biopsy in both males and females.

417 Background: Neoadjuvant chemotherapy (NC) improves survival in MIBC pts regardless of local management-cystectomy or chemoradiation. NC use with chemoradiation has been limited as these pts are often elderly, frail and cisplatin-ineligible. But, as more fit, cisplatin-eligible pts opt for chemoradiation it is critical that we re-evaluate the feasibility and outcomes of giving NC followed by chemoradiation. Methods: We reviewed 25 MIBC pts with cT 2-4 N 0-1 M 0, undergoing chemoradiation between 2008-2014 at the Princess Margaret and Durham Regional Cancer Centers. All pts received NC with gemcitabine-cisplatin (2-4 cycles) then external beam radiation (median dose 60Gy) with concurrent cisplatin. Toxicities were recorded using CTCAE v 3.0. Response and outcomes were assessed by cystoscopy and imaging. Median follow-up was 29.7 mos and 6 pts had < 2 yrs of follow-up. Kaplan Meier analysis was used for survival. Results: Main reasons for a chemoradiation approach were pt preference 15/25 (60%) and comorbidities 10/25(40%). At diagnosis, median age was 69 (49-85), 76% were male, all were ECOG PS 0/1, median CrCl was 58.5 ml/min and 7/25 (28%) had hydronephrosis. CIS was seen in 10/25 (40%), LVI in 3/25 (12%) and node positivity in 2/25 (8%). All but 1 patient completed planned NC, where main Grade 3/4 toxicities as expected were neutropenia and infection. All planned radiotherapy and 83% of planned concurrent chemotherapy was given. Maximal TURBT was done in 76%. Cystoscopically post NC, 12/15 (80%) had a CR, 1/15 (7%) had CIS, and 2/15 (13%) had residual disease. Of the 12 pts with a CR, radiologically 4/12 had a CR, 2/12 had a PR and 6/12 had SD. Four pts required salvage cystectomy for local recurrence, 4 pts developed metastases and have died. Median OS was not reached, but the 2 yr OS rate was 73.8% (95% CI 50.3-87.4%). Conclusions: NC followed by chemoradiation, showed cystoscopic CR rates of 80% post NC and 2 yr OS rates of 73.8% suggesting this approach should be considered in cisplatin-eligible MIBC pts undergoing chemoradiation. Comparing outcomes between matched MIBC pts receiving NC and then chemoradiation or cystectomy also appears warranted.

In the present study we determined the association of angiotensin converting enzyme (ACE) and plasminogen activator inhibitor-1 (PAI-1) gene polymorphisms with diabetic retinopathy (DR) and its sub-clinical classes in Pakistani type 2 diabetic patients. A total of 353 diabetic subjects including 160 DR and 193 diabetic non retinopathy (DNR) as well as 198 healthy controls were genotyped by allele specific polymerase chain reaction (PCR) for ACE Insertion/Deletion (ID) polymorphism, rs4646994 in intron 16 and PAI-1 4G/5G (deletion/insertion) polymorphism, rs1799768 in promoter region of the gene. To statistically assess the genotype-phenotype association, multivariate logistic regression analysis was applied to the genotype data of DR, DNR and control individuals as well as the subtypes of DR. The ACE genotype ID was found to be significantly associated with DR (p = 0.009, odds ratio (OR) 1.870 [95% confidence interval (CI) = 1.04-3.36]) and its sub-clinical class non-proliferative DR (NPDR) (p = 0.006, OR 2.250 [95% CI = 1.098-4.620]), while PAI polymorphism did not show any association with DR in the current cohort. In conclusion in Pakistani population the ACE ID polymorphism was observed to be significantly associated with DR and NPDR, but not with the severe form of the disease i.e. proliferative DR (PDR).

Introduction: Mantle cell lymphoma (MCL) often follows an aggressive course and remains incurable with standard therapies. First-line chemotherapy followed by consolidation with high-dose chemotherapy (HDT) and autologous stem cell transplant (ASCT) has become a standard of care in eligible patients (pts). As relapse remains the main cause of treatment failure, strategies such as intensifying induction therapy with high-dose cytarabine or adding rituximab maintenance (RM) have been tested to reduce the relapse rate (RR) post-ASCT. We evaluated the effect of the addition of cytarabine and RM on the outcome of pts undergoing ASCT. Methods: We conducted a retrospective analysis of consecutive MCL pts who underwent ASCT after first-line chemotherapy at the Princess Margaret Cancer Centre between 2000-2013. Pts received induction with CHOP, RCHOP, or RCHOP alternating with RDHAP (RCHOP/RDHAP), followed by HDT with or without total body irradiation (TBI). All pts had a documented response to induction using Cheson 1999 criteria. After ASCT, pts received maintenance with single-agent rituximab 375 mg/m2 or were simply observed. Results: 98 MCL pts were treated: median age was 56 years (36-66), 15 pts (15%) had blastoid or pleomorphic subtype, 85 pts (87%) had stage IV disease. MIPI was high risk in 18 pts (19%). Induction therapy: CHOP 14 pts (14%), RCHOP 57 (58%), and RCHOP/RDHAP 27 (28%). After induction CR was obtained in 44%, PR in 56% pts. CR rates were: CHOP 7 (50%), RCHOP 25 (44%), RCHOP/RDHAP 12 (44%) (P=ns). 89% pts had collected > 5*106 CD34/kg after RCHOP, and 78% after RCHOP/RDHAP (P=ns). Overall 66/98 pts (67%) had > 5*106 CD34/kg collected with 1 apheresis (Table 1). HDT was melphalan+etoposide for 63% pts, cytarabine+melphalan for 31% pts; 77 (79%) also received TBI. Median time from diagnosis to ASCT was 7.5 months (2.5, 33.4). Post-ASCT responses: CR 92 pts (94%), PR 4 (4%), 2 (2%) PD. Median time to ANC ≥0.5 were 10 days (CHOP), 11 days (RCHOP), and 11 days (RCHOP/RDHAP), while median days to PLT≥20 were 9 (CHOP), 11.5 (RCHOP), and 13 (RCHOP/RDHAP). Post-ASCT, 31% of pts had normal blood counts at 3 months which improved to 52% at 1 year post-ASCT. Maintenance data were available for 95/98 pts. RM was given to 72 pts (74%). Median follow-up from date of transplant for the entire cohort was 3.22 years (range 0.7 - 14.1). The 2-year and 5-year PFS were 85.8% (76.7-91.5) and 52.2% (37.7-64.7), respectively. 32 pts relapsed after ASCT (32.65%). Relapse occurred in 3 (11%) pts after RCHOP/RDHAP, 19 (33%) after RCHOP, and 10 (71%) after CHOP. Median time to relapse was 9 years (95%CI: 4.7-NR). 2-year and 5-year RR were 14.54% and 41.65%, respectively. Median OS was 9.15 years (95%CI: 7.3-NR), 2-year OS was 88.8% (80.2-93.8), and 5-year OS was 74.9% (61.7-84.2%). For patients observed without treatment post-ASCT, median PFS was 2.87 years (1.22-4.63) and median OS 5.19 years (1.66-NR), while for those receiving RM, PFS was 9.06 years (4.97-NR, p Conclusions: Response rate and PFS were similar between different induction regimens. The outcomes of responding pts following ASCT appear superior to previous strategies. Our patients enjoyed a very long PFS and median OS is surprisingly long as well. Within the limits of a retrospective study, our data support the use of rituximab maintenance, showing a significant benefit in both PFS and OS. Table 1. ASCT data Stem cell collection CHOP RCHOP RCHOP/RDHAP P value Pts collecting > 2x106 /Kg CD34+ cells in 1 day 4/14 (29%) 44/57 (77%) 17/27 (67%) 0.002 Pts collecting 2-5 x106 /Kg CD34+ cells N/A 6/57 (11%) 6/27 (22%) Pts collecting >5 x106 /Kg CD34+ cells N/A 51/57 (89%) 21/27 (78%) 0.153 Engraftment median (range) Days to ANC ≥ 0.5 10 (9,11) 11 (9, 12) 11 (9, 12) Pts with ANC ≥ 0.5 ≤ 11 days 13/13 (100%) 45/52 (87%) 21/25 (84%) 0.33 Days to PLT ≥ 20 9 (7, 15) 11.5 (9, 17) 13 (9, 26) Pts with PLT ≥ 20 ≤ 12 days 12/13 (92%) 37/52 (71%) 8/25 (32%) 0.0001* Days from ASCT to discharge 13 (11,26) 14 (11,30) 13 (11,23) PTs requiring RBC Transfusions 11 (79%) 43 (75%) 22 (81%) 0.821 Number of RBC Transfusions 2 (0, 4) 2 (0, 7) 3(0, 6) Pts requiring PLT Transfusions 12 (86%) 52 (93%) 25 (93%) 0.759 Number of PLT Transfusions 1 (0, 4) 2 (1, 9) 3 (1, 5) * comparison CHOP Vs R-CHOP: not significant, p=0.113 Legend. ASCT: autologous stem cell transplant; Pts: patients; ANC: absolute neutrophils count; PLTs: platelets; RBC: red blood cells. Disclosures Kuruvilla: Hoffmann LaRoche: Consultancy, Honoraria, Research Funding; Seattle Genetics: Consultancy, Honoraria; Gilead: Consultancy; Janssen: Consultancy, Honoraria; Merck: Honoraria; Bristol-Myers Squibb: Honoraria; Lundbeck: Honoraria; Karyopharm: Honoraria.

Abstract 3496 Notch1 signaling is required at multiple stages of normal T-lymphocyte development. Notch1 is a transmembrane receptor that is physiologically activated when Notch ligands induce conformational changes that allow Notch1 cleavage by the intramembranous γ-secretase complex, releasing active intracellular Notch1 (ICN1) fragment from the plasma membrane. Activating NOTCH1 mutations are very frequent in human and mouse T-cell lymphoblastic leukemia/lymphoma (T-LL). Typically, these mutations promote ligand-independent Notch1 cleavage by γ-secretase or increase ICN1 stability by truncating the C-terminal PEST domain. Understandably, much effort has focused on elucidating mechanisms of normal and oncogenic Notch1 signaling. However, some studies suggest that the absence of NOTCH1 mutations portends a worse prognosis for human T-LL. Therefore, we set out to define signals that promote proliferation and survival of T-LL cells lacking activated Notch1. We used Western blotting to detect γ-secretase cleaved ICN1 protein in a cohort of 35 primary T-LLs that developed spontaneously in mice lacking the Ataxia telangiectasia mutated (Atm) tumor suppressor. We identified 3 ICN1 subgroups: 63% expressed PEST-truncated ICN1 (T-ICN1); 17% expressed non-truncated ICN1 (NT-ICN1); and 20% had undetectable ICN1 (UD-ICN1), most lacked Notch1 mRNA. We confirmed the difference in Notch transcriptional activity and functional dependence between the UD-ICN1 and T-ICN1 subgroups and then compared their gene expression profiles to define pathways unique to the UD-ICN1 group. Gene set enrichment analyses revealed that UD-ICN1 T-LLs expressed higher levels of Klf9 and other transcription factors associated with a highly proliferative stage of normal T-cell development. siRNA knock-down studies demonstrated that Klf9 promoted proliferation of UD-ICN1 but not T-ICN1 T-LL cells. Collectively, these data demonstrate that Klf9 can regulate proliferation of Notch-independent T-LLs and suggest that Klf9 may provide a novel therapeutic target for human T-LLs lacking activating NOTCH1 mutations. Disclosures: No relevant conflicts of interest to declare.

Normal distribution based discriminant methods have been used for the classification of new entities into different groups based on a discriminant rule constructed from the learning set. In practice if the groups are not homogeneous, then mixture discriminant analysis of Hastie and Tibshirani (J R Stat Soc Ser B 58(1):155–176, 1996) is a useful approach, assuming that the distribution of the feature vectors is a mixture of multivariate normals. In this paper a new logistic regression model for heterogenous group structure of the learning set is proposed based on penalized multinomial mixture logit models. This approach is shown through simulation studies to be more effective. The results were compared with the standard mixture discriminant analysis approach using the probability of misclassification criterion. This comparison showed a slight reduction in the average probability of misclassification using this penalized multinomial mixture logit model as compared to the classical discriminant rules. It also showed better results when applied to practical life data problems producing smaller errors.

SummaryFrom an shRNA screen, we identified ClpP as a member of the mitochondrial proteome whose knockdown reduced the viability of K562 leukemic cells. Expression of this mitochondrial protease that has structural similarity to the cytoplasmic proteosome is increased in leukemic cells from approximately half of all patients with AML. Genetic or chemical inhibition of ClpP killed cells from both human AML cell lines and primary samples in which the cells showed elevated ClpP expression but did not affect their normal counterparts. Importantly, Clpp knockout mice were viable with normal hematopoiesis. Mechanistically, we found that ClpP interacts with mitochondrial respiratory chain proteins and metabolic enzymes, and knockdown of ClpP in leukemic cells inhibited oxidative phosphorylation and mitochondrial metabolism.