Academic literature on the topic '378.091.12:33-051]:378.22 (043.5)'

Abstract Abstract 5259 In this study, we attempted to determine the performance of the soluble fibrin monomer complex (SFMC) test in comparison to known activation markers, such as prothrombin fragment 1.2 (PF1.2), thrombin antithrombin complex (TAT) and d-dimer (DD). We have used the STA(R)-Liatest FM and STA(R)-Liatest D-Di kits to measure the SFMC and d-dimer levels. A total of 25 patients were included in this study. The data and demographic information are shown in the table below. ID GENDER AGE FM TAT PF1.2 D-dimer 1 M 66 0.42 2.3 1988 0.05 2 F 31 27.93 >60.0 >12000 2.39 3 F 24 3.41 14.1 1367 0.48 4 F 61 2.22 7.7 335 0.63 5 F 18 4.61 6.9 940 0.22 6 F 38 156.52 43.5 >12000 10.37 7 F 20 0.42 2.2 149 0.66 8 F 21 0.42 2.4 143 0.24 9 M 44 >150 15.4 11043 5.5 10 F 42 3.4 <2.0 134 0.24 11 M 46 0 4.8 271 0.3 12 M 76 90.73 29.4 1712 >20.0 13 F 61 3.41 3.8 484 0.52 14 M 1 0.1 3.8 116 0.33 15 F 33 4.2 37 6346 2.03 16 F 35 3.2 2.6 325 0.09 17 F 23 1.1 4.7 232 0.2 18 M 64 2.3 2.6 189 0.15 19 M 1 1.7 2.9 253 0.49 20 M 64 2.2 33.2 451 2.43 21 M 60 1.6 2.9 126 0.42 22 F 55 2.2 2.6 189 0.97 23 F 65 1.3 2.3 79 0.25 24 F 39 3.6 8.1 222 0.18 25 M 2 4.8 <2.0 109 0.53 Reference range >6.0 ug/mL <4 mcg/L 41–372 pmol/L <0.45 ug/mL The data indicate that in 16 out 25 patients, the SFMC levels are correlated with TAT and PF1.2. Only 3 out of 25 patients have elevated TAT and PF1.2 with normal SFMC levels. Therefore, our data would tend to indicate that SFMC level could be used as an activation marker to assess an ongoing prothrombotic process. In order to more fully determine the efficacy of the SFMC assay, we will obtain a detailed clinical history for the cases shown above. Disclosures: No relevant conflicts of interest to declare.

Abstract Background: Congenital adrenal hyperplasia due to classic 21-hydroxylase deficiency (21OHD) causes cortisol insufficiency and androgen excess. A phase 2 trial of crinecerfont, a CRF1 receptor antagonist, in 18 adults with 21OHD showed prominent decreases in ACTH, 17-hydroxyprogesterone, and androstenedione (A4), and in women, testosterone (T), after 14 days of treatment. In men with 21OHD, T derives from both adrenals and testes; in poor disease control, A4/T ratio is elevated due to disproportionately increased adrenal A4 production and decreased testicular T production. We sought to determine the impact of crinecerfont on both adrenal and gonadal androgen production in men with 21OHD in this phase 2 trial. Methods: A4 and T data were analyzed for 7 men who completed 1 or more of 4 oral dosing regimens: Cohort 1, 50 mg QHS, n=4; Cohort 2, 100 mg QHS, n=2; Cohort 3, 100 mg QPM, n=5; and Cohort 4, 100 mg BID, n=3 (14 total treatment periods). Mean 0600-1000 4-hour morning window (M4hMW) and mean 24-hour (M24h) A4, T, and A4/T ratios were analyzed from serial serum samples at baseline and on day 15. Results: Dose-dependent reductions in M4hMW A4 were observed [median (range)] in men, consistent with previously presented data in all subjects:Cohort 1: -21% (-84 to -12%);Cohort 2: -37% (-51% to -23%);Cohort 3: -43% (-85% to +140%);Cohort 4: -62% (-90% to -33%). In contrast, M4hMW T showed inconsistent changes [median (range)]: Cohort 1: +18% (-40% to +82%);Cohort 2: -4% (-4.3% to -3.8%);Cohort 3: +9% (-11 to +24%);Cohort 4: +9% (-3% to +27%). Thus, M4hMW A4/T ratios decreased with dose. Values at baseline, on day 15, and percent changes [median (range)] were, respectively:Cohort 1: 0.9 (0.3–2.6), 0.6 (0.1–2.1), -26% (-91% to +23%);Cohort 2: 5.0 (4.8–5.2), 3.3 (2.5–4.2), -35% (-49% to -20%);Cohort 3: 0.6 (0.1–6.9), 0.3 (0.1–2.7), -54% (-85% to +178%);Cohort 4: 3.9 (0.6–5.9), 0.4 (0.3–2.1), -65% (-92% to -31%). M24h A4/T ratios similarly declined in all cohorts. Values at baseline, on day 15, and percent changes [median (range)] were, respectively:Cohort 1: 1.0 (0.3–2.3), 0.4 (0.1–1.9), -33% (-92% to +2%);Cohort 2: 4.3 (3.8–4.9), 2.7 (2.4–3.0), -36% (-51% to -22%);Cohort 3: 0.5 (0.1–4.7), 0.4 (0.1–2.4), -59% (-78% to +310%);Cohort 4: 3.2 (0.4–4.1), 0.4 (0.3–1.7), -58% (-89% to -31%). Conclusions: Following crinecerfont therapy, A4 and A4/T decreased in a dose-dependent manner in men with 21OHD. In contrast to reductions in T observed in women with 21OHD, T did not change consistently and rose in some men. Preserved T values despite marked A4 reductions suggests testicular T production increased during crinecerfont therapy, perhaps due to release of gonadotropin suppression from adrenal-derived androgens. Long term studies are needed to determine if crinecerfont treatment improves additional measures of testicular function in men with 21OHD. Reference: RJ Auchus, et al. J Endocr Soc 2020;4(Suppl 1):OR25-03.

Abstract Myelodysplastic syndromes(MDS) are a heterogeneous group of myeloid neoplasms characterized by bone marrow failure with peripheral cytopenia and morphologic dysplasia in one or more of hematopoietic cell lineages. Chromosomal abnormalities, detected in 40–70% patients with primary MDS, are presented heterogeneous due to the disparities in race and environment. In the current study, 351 adult patients in our institution were retrospectively analyzed to clarify features of karyotype in Chinese primary MDS. There were 237 cases (67.5%) demonstrated karyotypic abnormalities including numeral changes alone(99 cases, 41.7%), structural changes alone(70 cases,29.5%) and both changes(68 cases,28.8%). Single, double and complex (more than two independent aberrations) anormaly were 130 cases(54.8%),54 cases(22.8%), 53 cases(22.4%), respectively. There were 3 cases(1.3%) with tetraploid, 1 case(0.4%)with multiploid,1 case(0.4%) with hyperdiploid and 3 cases(1.3%) with hypodiploid. Changes of aneuploid or anomaly of chromosomal arm could be detected in all of the 24 chromosomes and the aberrations frequently seen were +8, −20/20q-/del(20)(q11–13), −7/7q-/del(7)(q21–32), −5/5q-/del(5)(q13–33), −18, −11/11q-/del(11) (q23–25), +21, −Y, −21, −10, −16, −22, +9, del(12) (p12) in order. The incidence of −5/5q-/del(5)(q13–33)(5.1%)was lower than western countries(8.7–23.4%)and 5q-syndrome was even less (0.3%). The incidences of +8(19.1%) and −20/20q-/del(20)(q11–13) (9.4%) were higher than western countries(1.2–7.0%, 2.0– 3.5%, respectively). Chromosome translocations could be detected in 31 cases(13.1%) and among them 12 translocations have not been reported in MDS. In addition, i(17)(q10) could be detected in 9 cases(3.8%)in which 6 cases were single anormaly. Chromosomal duplication presented in 7 cases(3.0%) in which 4 cases involved chromosome 1. According to IPSS chromosomal prognositic classification, the incidence of poor-risk karyotypes was increasing as following orders: RA, RARS and 5q-syndrome;RCMD and RCMDRS;RAEB-I;RAEB-II and the difference was statistically significant(χ2 = 59.169, P &lt; 0. 001). Among the 178 patients with follow-up data, the median follow-up time was 14.5(1–131)months and the median OS was 36 (95% confident interval(CI) 25–46) months with 2 and 5 year of OS 58.7% and 38.0%, respectively. According to IPSS chromosomal prognositic classification, the median OS of patients with good, intermediate and poor-risk cytogenetic subgroups were 51(95%CI 25–77),35(95%CI 5–65),13(95%CI 9–17)months respectively and the difference of OS was statistically significant by log rank test(P=0. 004). According to NN-AN-AA classification, the median OS of patients with NN, AN, AA were 51(95%CI 24–78),36(95%CI 0.3–71), 23(95%CI 10–35)month respectively and the difference of OS was statistically significant by log rank test(P =0. 039). In summary, features of chromosomal abnormalities in Chinese patients with primary MDS were not the same with that in western countries. Cytogenetical abnormalities validate vigorously malignant texture of MDS and are considered to be of great importance in revealing pathogenesis, determinating diagnosis and evaluating prognosis in MDS.

Background The COVID-19 pandemic resulted in a rapid shift from center-based rehabilitation to telerehabilitation for chronic respiratory disease and lung transplantation due to infection control precautions. Clinical experience with this delivery model on a large scale has not been described. Objective The aim of this study is to describe usage and satisfaction of providers and lung transplant (LTx) candidates and recipients and functional outcomes following the broad implementation of telerehabilitation with remote patient monitoring during the first wave of the COVID-19 pandemic. Methods This study was a program evaluation of providers, LTx candidates, and early LTx recipients who used a web-based, remote monitoring app for at least four weeks between March 16 and September 1, 2020, to participate in telerehabilitation. Within-subjects analysis was performed for physical activity, Self-efficacy For Exercise (SEE) scale score, aerobic and resistance exercise volumes, 6-minute walk test results, and Short Physical Performance Battery (SPPB) results. Results In total, 78 LTx candidates and 33 recipients were included (57 [51%] males, mean age 58 [SD 12] years, 58 [52%] with interstitial lung disease, 34 [31%] with chronic obstructive pulmonary disease). A total of 50 (64%) LTx candidates and 17 (51%) LTx recipients entered ≥10 prescribed exercise sessions into the app during the study time frame. In addition, 35/42 (83%) candidates agreed the app helped prepare them for surgery and 18/21 (85%) recipients found the app helpful in their self-recovery. The strongest barrier perceived by physiotherapists delivering the telerehabilitation was patient access to home exercise and monitoring equipment. Between the time of app registration and ≥4 weeks on the waiting list, 26 LTx candidates used a treadmill, with sessions increasing in mean duration (from 16 to 22 minutes, P=.002) but not speed (from 1.7 to 1.75 mph, P=.31). Quadriceps weight (pounds) for leg extension did not change (median 3.5, IQR 2.4-5 versus median 4.3, IQR 3-5; P=.08; n=37). On the Rapid Assessment of Physical Activity questionnaire (RAPA), 57% of LTx candidates scored as active, which improved to 87% (P=.02; n=23). There was a decrease in pretransplant 6-minute walk distance (6MWD) from 346 (SD 84) meters to 307 (SD 85) meters (P=.002; n=45) and no change in the SPPB result (12 [IQR 9.5-12] versus 12 [IQR 10-12]; P=.90; n=42). A total of 9 LTx recipients used a treadmill that increased in speed (from 1.9 to 2.7 mph; P=.003) between hospital discharge and three months posttransplant. Quadriceps weight increased (3 [IQR 0-3] pounds versus 5 [IQR 3.8-6.5] pounds; P<.001; n=15). At three months posttransplant, 76% of LTx recipients scored as active (n=17), with a high total SEE score of 74 (SD 11; n=12). In addition, three months posttransplant, 6MWD was 62% (SD 18%) predicted (n=8). Conclusions We were able to provide telerehabilitation despite challenges around exercise equipment. This early experience will inform the development of a robust and equitable telerehabilitation model beyond the COVID-19 pandemic.

Abstract Presence of internal tandem duplication in the FMS-like tyrosine kinase 3 gene(s) (FLT3-ITD) is associated with poor outcome among patients with cytogenetically normal acute myelogenous leukemia (CN-AML). Patients carrying a higher FLT3-ITD allelic burden have worse outcome. Allogeneic stem cell transplantation (SCT) in first remission (CR1) is reported to improve survival in this poor risk patients. We investigated the impact of FLT3-ITD allelic burden, number of FLT3-ITD mutations and SCT with a matched related or unrelated donor in CR1 in newly diagnosed FLT3-ITD mutated intermediate cytogenetic risk AML patients diagnosed between July 2000 and November 2013, who achieved CR1 with induction therapy. In order to reduce the selection bias, only patients who were alive and free of AML for at least 4 months were included in the analysis (median time to SCT was 4.5 months). The study group had 169 patients and 48 of 169 had SCT in CR1. Patients in the transplant group were younger than those in the non-transplant group (median age 55 v 62 years, p=0.001). As presented in Table1, there was no significant difference in the distribution of gender, white blood cell and platelet counts, bone marrow (BM) blast number, the proportion with CN, the level of FLT3-ITD allele level and number of FLT3-ITD mutations at diagnosis between the two groups. The median follow-up of survivors in the transplant and no transplant groups were similar (30.7 vs. 34.5 months). Overall, SCT in CR1 was associated with longer relapse-free survival (RFS) compared to no-transplant approach. The 3-year RFS from CR1 for transplant patients was 46.3% (95% confidence interval (CI),33.3%-64.2%) compared to 17.5% (95% CI,11.5%-26.4%) for non-transplant patients (P<0.001) (Figure 1). This was largely due to a higher cumulative relapse incidence (RI) of 68% in the non-transplant patients compared to 41% who received SCT (P<0.001). Overall survival (OS) from CR1 was also different between the groups (P=0.0003), with 3-year estimates of 53.5% (95% CI,39.6%-72.3%) for transplant patients compared to 24.3% (95% CI, 17.3%-34.2%) for no-transplant patients. The non-relapse mortality rate of 13% observed with SCT was similar to 15% observed in non-transplant patients (P=0.86). Higher WBC and FLT3-ITD alleleic burden at diagnosis were poor prognostic factors associated with decreased RFS and OS in univariate analyses. Older age decreased OS but not RFS. For multivariate regression, we excluded WBC count at diagnosis from the analysis as high WBC was associated with FLT3-ITD allelic burden at diagnosis. Multivariate regression models showed similar results with SCT in CR1 leading to improved RFS and OS while older age and higher FLT3-ITD allelic burden lost their prognostic significance as shown in Table 2. Our results indicate that SCT in CR1 AML FLT3-ITDmut patients is associated with a longer RFS, largely due to a reduction in the RR. The main reason of treatment failure remains to be relapse and strategies incorporating FLT3-ITD inhibitors before and after SCT may lead to a significant improvement in RFS of this high risk group. Table 1: Patient and disease characteristics All patients Transplant group Non-transplant group p Median age, years (IQR) 59 (50-68) 55 (47-62) 62 (51-70) 0.001 Age <60 85 (50.3%) 30 (62.5%) 55 (45.5%) 0.06 Female 82 (48.5%) 25 (52.1%) 57 (47.1%) 0.6 Diagnosis after 2008 106 (62.7%) 38 (79.2%) 68 (56.2%) 0.01 Median WBC at diagnosis (IQR) 11.6 (3.8-29.7) 9 (3-40) 12 (4-28) 0.98 Median platelets at diagnosis, (IQR) 45 (25-74) 45 (22-84) 45 (25-71) 0.99 Median BM blast count at diagnosis, (IQR) 43(12-74) 32 (6-64) 46 (13-75) 0.2 Diagnostic cytogenetics Diploid 128 (75.7%) 33 (68.8%) 95 (78.5%) Other abnormalities 36 (21.3%) 12 (25%) 24 (19.8%) 0.4 Unknown 5 (2.9%) 3 (6.3%) 2 (0.2%) Median FLT3/ITD allelelic burden, (IQR) 0.34 (0.1-0.47) 0.30 (0.04-0.47) 0.35 (0.12-0.49) 0.3 FLT3/ITD allele ratio <0.3 75 (44.4%) 24 (50%) 51 (42.1%) 0.5 Number of FLT3/ID mutations 1 130 (76.9%) 35 (72.9%) 95 (78.5%) 2 21 (12.4%) 9 (18.8%) 12 (9.9%) 3 12 (7.1%) 3 (6.3%) 9 (7.4%) 0.4 Unknown 6 (3.6%) 1 (2.1%) 5 (4.1%) Table 2: Multivariate regression models for RFS and OS Variables RFS OS HR (95%CI) p HR (95%CI) p Log (FLT3-ITD allele level at diagnosis) 1.12(0.9-1.3) 0.14 1.13 (0.95-1.33) 0.16 HSCT No 1.0 1.0 Yes 0.38 (0.24-0.60) <0.001 0.43 (0.26-0.70) 0.001 Figure 1: Kaplan-Meier estimates of leukemia-free survival by transplant groups Figure 1:. Kaplan-Meier estimates of leukemia-free survival by transplant groups Disclosures Kantarjian: ARIAD, Pfizer, Amgen: Research Funding.

Abstract INTRODUCTION: More active high-dose regimens are needed for refractory or poor-risk relapsed Hodgkin's lymphomas (HL), where BEAM offers poor results. Post-BEAM maintenance treatment with brentuximab vedotin (BV) x 48 weeks has recently been shown in the AETHERA trial to prolong progression-free survival (PFS) compared to placebo (2-year PFS 63% vs. 51%). We previously developed a regimen of infusional gemcitabine combined with busulfan and melphalan (Gem/Bu/Mel) pursuing inhibition by Gem of DNA damage repair. The encouraging results we saw in HL patients led us to conduct a phase 2 trial of Gem/Bu/Mel in HL patients at high risk of post-ASCT relapse. METHODS: HL patients ages 12-65 with ≥1 of the following criteria were eligible: Persistent active disease after 1st-line chemotherapy, CR1 < 1 year, or extranodal disease at relapse/PD. Gem was administered as a loading dose of 75 mg/m2 followed by infusion at a fixed dose rate of 10 mg/m2/min over 4.5 hours on days -8 and -3 (total daily dose of 2,775 mg/m2). Each Gem infusion was immediately followed by the corresponding dose of Bu or Mel. Bu was administered intravenously from days-8 to -5 targeting a daily AUC of 4,000. Mel was infused at 60 mg/m2/day on days -3 and -2. ASCT was on day 0. Post-HDC involved field radiotherapy (IFRT) was considered to lesions >5 cm at the time of HDC or persistently PET+ at the 1-month post-HDC evaluation. The trial had 80% power to detect a 2-year PFS increase from 50% to 65%.The concurrent BEAM cohort included all patients eligible for this trial who received BEAM off study due to no financial coverage for ASCT in a trial or patient/physician preference. RESULTS: Eighty patients were enrolled on study between 6/11-04/15 (Table 1). There was no transplant-related mortality (TRM). The toxicity profile was manageable, including mucositis (49% G2, 40% G3), skin (22% G2, 11% G3), self-limited transaminase elevation (30% G2, 19% G3), and hyperbilirubinemia (24% G2, 19% G3) with no cases of VOD. There was 1 case of G2 pneumonitis and none of cardiac, renal or CNS toxicity. Neutrophils and platelets engrafted at median days +10 (8-12) and +12 (9-21), respectively. Eight patients received post-HDC IFRT to mediastinum ± neck ± sternum at 30.6-39.6 Gy, starting on median day +42 (41-53) post-HDC, which was well tolerated. No patients received maintenance BV. Table 1. Patient characteristics Variable Study file (N=80) Concurrent BEAM cohort (N=31) P Median age (range) 31 (13-65) 39 (23-65) 0.02 Primary refractory / poor-risk relapse 41% / 59% 37% / 63% 0.6 # prior relapses 1 80% 70% 0.3 >1 20% 30% Median # prior chemotherapy lines (range) 2 (2-6) 2 (2-7) 0.3 Prior disease-free interval (months) <6 56% 60% 0.7 6-12 24% 17% >12 20% 23% Prior xRT 21% 27% 0.6 Relapse within prior xRT field 10% 3% 0.4 Extranodal relapse/PD 36% 53% 0.08 B symptoms at relapse/PD 11% 10% 0.8 Bulky relapse (any lesion >5 cm) 39% 17% 0.02 # risk factors (primary refract/CR1<1 yr, extranodal relapse, or B symptoms) 1 74% 77% 0.2 2 26% 17% 3 0% 6% Prior BV % 14% 25% 0.1 CR 36% 50% 0.1 PR 36% 0% No response (NR) 26% 50% PET+ at HDC 32% 7% 0.003 Status at HDC: CR/ PR / NR 68% / 24% / 8% 93% / 7% / 0% 0.01 At median follow-up of 33 mo (4-50) there have been 25 relapses following Gem/Bu/Mel, at median 6 (2-22) mo post-HDC (only 3 relapses after 12 mo). On univariate analyses, PET+ at HDC and primary refractoriness correlated with worse EFS (Table 2). On multivariate analyses, PET+ was an independent adverse predictor. Table 2. Prognostic analyses Variable Univariate analyses Multivariate analyses 2-yr EFS P HR (95% CI) P Yes No PET+ 37% 82% 0.00008 3.8 (1.7-8.9) 0.001 Primary refractory 51.5% 81% 0.006 2.2 (0.9-5.1) 0.06 >1 relapse 50% 73% 0.08 B symptoms 55.6% 70.4% 0.2 Bulky relapse 67% 72% 0.2 Extranodal 67% 72% 0.4 The BEAM cohort included 31 patients treated between 06/11-04/15 (Table 1) with no BV maintenance. There were fewer cases of PET+ tumors at HDC (P=0.003) and of bulky relapses (P=0.02) than the Gem/Bu/Mel file, but was matched for the other risk factors. It had no TRM. Despite a higher number of PET+ tumors at HDC, the Gem/Bu/Mel file had significantly superior 2-year PFS (65% vs. 51%, P=0.03) and 2-year OS (95.5% vs. 70%, P=0.001) than the BEAM cohort. CONCLUSIONS: Gem/Bu/Mel without maintenance BV was safe and effective in patients with refractory or poor-risk relapsed HL, with comparable results to those from the AETHERA trial using BEAM and maintenance BV. A randomized trial is necessary to compare Gem/Bu/Mel and BEAM. Figure 1. Figure 1. Disclosures Off Label Use: Gemcitabine, busulfan and melphalan are not FDA approved at high doses for Hodgkin's lymphoma. Alousi:Therakos, Inc: Research Funding. Andersson:Otsuka Research and Development, Inc.: Consultancy. Fanale:Merck: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; BMS: Research Funding; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Takeda: Honoraria, Research Funding; Infinity: Membership on an entity's Board of Directors or advisory committees; Spectrum: Membership on an entity's Board of Directors or advisory committees; Seattle Genetics: Honoraria, Research Funding; Genentech: Research Funding; Medimmune: Research Funding; Novartis: Research Funding; Bayer: Membership on an entity's Board of Directors or advisory committees; Amgen: Membership on an entity's Board of Directors or advisory committees; Molecular Templates: Research Funding; ADC Therapeutics: Research Funding; Onyx: Research Funding; Gilead: Research Funding.

Abstract Background BTZ is approved for the treatment of MM via either SC or IV administration. The observed non-inferior efficacy (overall response rate after 4 cycles) and tolerability with SC administration of BTZ as reported in relapsed/refractory MM (Moreau et al Lancet Oncol 2011) has the potential to impact treatment outcomes in the clinical setting. However, there is a paucity of comparative effectiveness data on SC vs IV BTZ as initial therapy in MM in clinical practice. The main objective of this study was therefore to evaluate the treatment outcomes in NDMM pts who received SC vs IV BTZ in the US community oncology setting. Methods NDMM pts aged ≥18 yrs who received BTZ-based treatment within the McKesson Specialty Health/US Oncology Network between Jan 1 and Dec 31, 2012, and had ≥2 MM-related visits and ≥6 mos' follow-up through Jul 2014, were evaluated. Data were collected via the iKnowMed electronic health records database and supplemented with medical chart reviews. Cohorts stratified by route of administration were matched 2:1 as the Moreau study, and by age, gender, practice region, and performance status. Chi-square analysis (categorical variables), t-tests/non-parametric Mann-Whitney tests (continuous variables), and Kaplan-Meier methodology were employed. Univariate and multivariate Cox models adjusted for clinically relevant covariates were used for associations between cumulative BTZ dose and treatment outcomes. Results 372 NDMM pts who received BTZ were included; 248 received SC and 124 IV BTZ. Similar baseline demographics and clinical characteristics were observed in the SC and IV groups (Table 1). Median BTZ treatment duration was 4.4 and 4.1 mos in the SC and IV groups, respectively (p = 0.21). Among pts receiving SC and IV BTZ, 10% and 13% had dose reductions in the first 16 wks (p = 0.41) and the median time to dose reduction was 49 d in both groups. While observed median relative dose intensities were similar between the SC and IV groups (1.0 [range 0.67-1.2] vs 1.0 [0.5-1.3]; p = 0.89), a higher median cumulative BTZ dose of 25.8 mg/m2 (1.3-208.6) was observed in SC pts, vs 22.9 mg/m2 (1.3-105.9) in IV pts (p = 0.08). When dichotomized by median cumulative BTZ dose (24.7 mg/m2), a numerically greater proportion of SC pts vs IV pts received the higher dose (52% vs 48%, p = 0.38). Higher cumulative dose was associated with longer PFS and better response rate at cycle 4 (12 wks) in multivariate analyses (Table 2). Treatment outcomes were comparable between SC and IV BTZ; 2-yr overall survival (OS) was 81% vs 78% (Fig 1); 2-yr progression-free survival (PFS) was 75% vs 70% (Fig 2). In 204 response-evaluable pts (of 372 in the overall population with a median follow up of 19.1 mos [range 0.1-30.1] in SC pts and 20.8 mos [0.3-30.8] in IV pts), complete response/very good partial response rate was 16% vs 22%. Documented rates of adverse events in the SC vs IV BTZ groups were 34% vs 28% neuropathy (p = 0.27), 13% vs 10% thrombocytopenia (p = 0.57), 10% vs 5% neutropenia (p = 0.09), and 25% vs 24% anemia (p = 0.87). Conclusions The observed non-inferior treatment outcomes between SC and IV BTZ in NDMM pts are consistent with findings from the phase 3 relapsed/refractory MM trial. In addition, SC vs IV BTZ was associated with a trend towards higher cumulative dose, which was associated with improved response rate and PFS. These findings support extended therapy for higher cumulative dosage and better treatment outcomes as demonstrated in a prior phase 3 clinical trial (Benboubker et al NEJM 2014). Table 1. NDMM Pt Baseline Demographics and Clinical Characteristics BTZ SC (N = 248) BTZ IV (N = 124) p value Median age at diagnosis, yrs (range) 66 (32-90+) 68 (36-88) 0.87 Age at diagnosis ≥65 yrs, % 57 60 0.55 Male % 56 56 1.00 IgG / IgA / light chain / other or unknown MM, % 53 / 21 / 22 / 4 41 / 30 / 23 / 6 0.28 Pts with available ISS stage I / II / III disease, % n = 145 22 / 43 / 34 n = 64 17 / 33 / 50 0.12 Peripheral neuropathy, % 4.4 3.2 0.78 Table 2. Regression analyses (RA) for association with cumulative BTZ dose, high vs low (> or < 24.7 mg/m2) Univariate analysis Multivariate analysis OR (95% CI) p value OR (95% CI) p value Objective response* at cycle 4 1.67 (0.86, 3.24) 0.13 2.20 (1.01, 4.80) 0.047 HR (95% CI) HR (95% CI) PFS† 0.41 (0.27, 0.62) <0.0001 0.34 (0.22, 0.54) <0.0001 *Logistic RA †Cox RA Disclosures Rifkin: Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees; Millennium Pharmaceuticals, Inc., Cambridge, MA, USA, a wholly owned subsidiary of Takeda Pharmaceutical Company Limited: Consultancy, Membership on an entity's Board of Directors or advisory committees; Onyx Pharmaceuticals: Consultancy, Membership on an entity's Board of Directors or advisory committees. Ma:Millennium Pharmaceuticals, Inc., Cambridge, MA, USA, a wholly owned subsidiary of Takeda Pharmaceutical Company Limited: Employment. Harrell:McKesson Specialty Health: Employment. Zhu:Millennium Pharmaceuticals, Inc., Cambridge, MA, USA, a wholly owned subsidiary of Takeda Pharmaceutical Company Limited: Employment. Dow:Millennium Pharmaceuticals, Inc., Cambridge, MA, USA, a wholly owned subsidiary of Takeda Pharmaceutical Company Limited: Employment; Takeda: Equity Ownership. Niculescu:Millennium Pharmaceuticals, Inc., Cambridge, MA, USA, a wholly owned subsidiary of Takeda Pharmaceutical Company Limited: Employment. Bonthapally:Millennium Pharmaceuticals, Inc., Cambridge, MA, USA, a wholly owned subsidiary of Takeda Pharmaceutical Company Limited: Employment.

Abstract Background: Prognosis for older patients with relapsed or refractory (R/R) acute myeloid leukemia (AML) is poor, with lower response rates and shorter survival compared with younger patients. Vosaroxin is a first-in-class anticancer quinolone derivative that is active in AML, is minimally metabolized, evades P glycoprotein receptor-mediated efflux, and has activity independent of p53 status. In the phase 3 VALOR trial, overall survival (OS) was significantly improved with vosaroxin plus cytarabine vs placebo plus cytarabine in a prespecified analysis in patients ≥ 60 years of age (median OS: 7.1 mo vs 5.0 mo, respectively; HR = 0.75 [95% CI: 0.62-0.92]; P = 0.0030). At the time of the primary analysis (September 2014), patients had been followed for a median of 24.4 months (as estimated by the reverse Kaplan-Meier method). Here, after a median follow-up of 39.9 months, we provide updated survival information for patients ≥ 60 years treated in the VALOR trial. Methods: In VALOR, patients with refractory or first relapsed AML were randomized 1:1 to receive cytarabine (1 g/m2 IV over 2 h, d 1-5) plus either vosaroxin (90 mg/m2 IV over 10 min, d 1, 4; 70 mg/m2 in subsequent cycles) or placebo. Eligible patients had refractory disease (persistent disease after induction, or first complete remission [CR1] lasting < 90 d) or were in first relapse (early relapse: CR1 lasting 90 d to 12 mo; late relapse: CR1 lasting 12 mo to 24 mo). Randomization was stratified by age (< 60, ≥ 60 years), disease status (refractory, early relapse, late relapse), and geographic location (US, non-US). Patients were followed for survival until death. At the time of the primary analysis (September 2014), 63/451 (14%) patients ≥ 60 years were alive and in continued follow-up. Results:As of Jan 22, 2016, 33/451 patients ≥ 60 years treated in the VALOR trial were alive and in continued follow-up: 23/226 (10.2%) in the vosaroxin/cytarabine arm and 10/225 (4.4%) in the placebo/cytarabine arm. Updated OS data in patients ≥ 60 years of age was consistent with the primary analysis, demonstrating a significant improvement with the addition of vosaroxin (Figure; HR = 0.75 [95% CI: 0.62-0.91]; P = 0.0017). The survival benefit associated with vosaroxin/cytarabine was durable, as demonstrated by the separation of the survival curves through 48 mo. At 24 and 36 mo after randomization, the estimated survival rate was twice as high with vosaroxin/cytarabine as with placebo/cytarabine in patients ≥ 60 years (17.0% vs 8.5% and 12.8% vs 6.0%, respectively). The OS benefit was consistent across various age subgroups in the population ≥ 60 years of age; in patients 60-64 years (n = 124), 65-74 years (n = 293), and 75-84 years (n = 34), vosaroxin/cytarabine treatment increased median survival by 2.9 mo (8.1 vs 5.2 mo; HR = 0.72 [95% CI: 0.49-1.06]), 2.0 mo (7.0 vs 5.0 mo; HR = 0.76 [95% CI: 0.60-0.97]), and 2.2 mo (5.5 vs 3.3 mo; HR = 0.72 [95% CI: 0.36-1.45]) over placebo/cytarabine treatment, respectively. An improvement in OS was observed among patients ≥ 60 years who received post-treatment transplantation (n = 91; 19.9 mo with vosaroxin/cytarabine vs 12.2 mo with placebo/cytarbine; HR = 0.70 [95% CI: 0.43-1.13]) and those who did not (n = 360; 5.3 mo vs 3.8 mo, respectively; HR = 0.75 [95% CI: 0.60-0.92]). When analyzed by disease status, the greatest OS benefit was observed in patients ≥ 60 years with refractory disease (n = 210; 6.7 mo with vosaroxin/cytarabine vs 3.8 mo with placebo/cytarabine; HR = 0.75 [95% CI: 0.57-1.00]) and those with early relapse (n = 154; 6.5 mo vs 3.9 mo, respectively; HR = 0.62 [95% CI: 0.44-0.87]). In patients ≥ 60 years with late relapse (n = 87), median OS was similar in both treatment arms (9.2 mo with vosaroxin/cytarabine vs 9.8 mo with placebo/cytarabine; HR = 1.06 [95% CI: 0.68-1.66]). Conclusions: After a median of 39.9 months of follow-up in the VALOR trial, the OS observed in patients ≥ 60 years of age treated with vosaroxin/cytarabine remains significantly improved versus placebo/cytarabine, with a separation of the survival curves through 48 months. The OS benefit is consistent among all older patients, even those ≥ 75 years of age, and is seen in patients with and without post-treatment transplantation. The updated survival data from VALOR continue to support the use of vosaroxin/cytarabine as a treatment option in patients ≥ 60 years of age with R/R AML. Disclosures Ravandi: BMS: Research Funding; Seattle Genetics: Consultancy, Honoraria, Research Funding. Ritchie:Novartis: Honoraria, Research Funding; Celgene: Speakers Bureau; Incyte: Speakers Bureau; Pfizer: Honoraria; Arian: Speakers Bureau. Vey:Sunesis: Honoraria. Strickland:Alexion Pharmaceuticals: Consultancy; Ambit: Consultancy; Baxalta: Consultancy; Boehringer Ingelheim: Consultancy, Research Funding; CTI Biopharma: Consultancy; Daiichi Sankyo: Consultancy; Sunesis Pharmaceuticals: Consultancy, Research Funding; Abbvie: Research Funding; Astellas Pharma: Research Funding; Celator: Research Funding; Cyclacel: Research Funding; GlaxoSmithKline: Research Funding; Karyopharm Therapeutica: Research Funding; Sanofi: Research Funding. Schiller:Incyte Corporation: Research Funding. Jabbour:ARIAD: Consultancy, Research Funding; Pfizer: Consultancy, Research Funding; Novartis: Research Funding; BMS: Consultancy. Erba:Ariad: Consultancy; Millennium Pharmaceuticals, Inc.: Research Funding; Celator: Research Funding; Novartis: Consultancy, Speakers Bureau; Amgen: Consultancy, Research Funding; Agios: Research Funding; Juno: Research Funding; Gylcomimetics: Other: DSMB; Astellas: Research Funding; Incyte: Consultancy, DSMB, Speakers Bureau; Seattle Genetics: Consultancy, Research Funding; Pfizer: Consultancy; Celgene: Consultancy, Speakers Bureau; Daiichi Sankyo: Consultancy; Sunesis: Consultancy; Jannsen: Consultancy, Research Funding. Pigneux:Agios: Consultancy, Honoraria; Sunesis: Consultancy, Honoraria. Horst:Amgen, Novartis, Pfizer, Gilead, Agios: Consultancy; Amgen, Regeneron: Research Funding; Celgene: Honoraria. Recher:Celgene, Sunesis, Amgen, Novartis: Membership on an entity's Board of Directors or advisory committees; Celgene, Sunesis, Amgen, Novartis, Chugai: Research Funding. Cortes:ARIAD: Consultancy, Research Funding; Bristol-Myers Squib: Consultancy, Research Funding; Novartis: Consultancy, Research Funding; Pfizer: Consultancy, Research Funding; Teva: Research Funding. Roboz:Agios, Amgen, Amphivena, Astex, AstraZeneca, Boehringer Ingelheim, Celator, Celgene, Genoptix, Janssen, Juno, MEI Pharma, MedImmune, Novartis, Onconova, Pfizer, Roche/Genentech, Sunesis, Teva: Consultancy; Cellectis: Research Funding. Craig:Sunesis: Employment, Equity Ownership. Ward:Sunesis Pharmaceuticals: Consultancy, Employment. Smith:Sunesis: Employment, Equity Ownership. Stuart:Agios: Research Funding; Sunesis: Consultancy, Honoraria, Other: Travel, Accomodations, Expenses, Research Funding; Incyte: Research Funding; Bayer: Research Funding; Celator: Research Funding; Astellas: Research Funding.

Green gram (Vigna radiata) is considered the chief legume in Pakistan. Thus, current study was conducted to examine the ameliorating effect of phytohormones pre-treatments under salt stress on proteome profile of green gram by sodium-dodecyl-sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The soluble green gram seedlings proteins were resolved on 4% stacking and 12% resolving gels. The SDS-PAGE resolved 24 polypeptide bands ranging from 200 to 17kDa. Among these, 12 out of 24 bands of proteins were essentials house-keeping or growth proteins of green grams. While, 120, 114.6, 51.8, 29.1, and 22.8 kDa bands were over-expressed under 50 to 350mM salt with phytohormones treatments. The others 104.5 kDa, 99.8 kDa, 95.3 kDa, 91.0 kDa, 55 kDa, 46 kDa, and 17kDa bands were related to the GAᴣ, IAA, and SA induced tolerance. Overall 120 kDa, 114.6 kDa, 104.5 kDa, 99.8, 95.3 kDa, 51.8 kDa, 29.1 kDa and 22.8kDa bands were first time identified in the current study. The information retrieved from NCBI protein database, the resolved peptides were principally belonging to 7S and 8S vicilin, 2S, 8S, 11S, and 16.5S globulins. It is determined that seed priming with SA enhanced tolerance in green gram by rapidly synthesizing stress alleviating peptides.Key word: Cluster analysis, dendrogram, mungbean, salt stress, SDS-PAGEINTRODUCTIONVarious world-wide health concerning organization recommended the use of high graded plant protein such as legumes to prevent the risk of metabolic disorder (Hou et al., 2019). Legumes are most important protein crop on the earth. Among the legumes, the green gram is the major pulses. Its seeds are rich in superior quality storage protein, which account 85% of the total protein while, another 15% have not been broadly studied (Yi-Shen et al., 2018). The soluble storage protein comprises of 60% globulins, 25% albumin and 15% prolamins. Globulins are further divided into 3.4% basic-type (7S), 7.6% legumin-type (11S), and 89% vicilin-type (8S) (Mendoza et al., 2001; Itoh et al., 2006). Other than proteins, the green gram seeds also contain starch, fiber, phenolic compound, saponins, vitamins, calcium zinc, potassium, folate, magnesium, manganese and very low in fat that made it meager man’s meat (Hou et al., 2019). It is also a good source of green manure and fodder (El-Kafafi et al., 2015). Its root has ability to fix 30 to 50 Kg/ha atmospheric nitrogen in the soil which is essential for maintaining soil fertility (Chadha, 2010). The green gram is the valuable and the major Rabi pulse crop of Pakistan. Its cultivation area in 2016-2017 was about 179,000 hectares with seed yield of 130,000 tones. In comparison during 2017-2018, it was cultivated on 161,800 hectares land with 118,800 tones seed yield (GOP, 2018). One of the reasons of this 9% decrease in both land and productivity is the shortage of irrigated land due to soil salinity. The salinity induce oxidative bust in the mungbean cells, caused by responsive oxygen species (ROS) such as hydrogen peroxide, singlet oxygen, hydroxyl radical and superoxide radical. The ROS create hindrance in various metabolic processes of plant via interacting with macromolecules like proteins (Alharby et al., 2016). However, phytohormones like gibberellic acid (GAᴣ), indole acetic acid (IAA), and salicylic acid (SA) take part in the biosynthesis of salt tolerance proteins under salinity. These salt tolerance proteins acclimate plants under salinity stress. Application of biotechnology plays a significant role in agriculture (Khan et al., 2017). Therefore, production of particular proteins under salt stress is a specific response of cell which can be analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). SDS-PAGE is the simple, valid, and cost-effective biochemical marker (Mushtaq et al., 2018). This marker has been widely used to determine the extent of evolutionary variations in crops (El-Kafafi et al., 2015).OBJECTIVES The present study was directed first time with the aim to investigate the toxic effect of sodium chloride (0-350 mM) and stress acclimation by pre-treatment of GAᴣ, IAA, and SA on the proteome profile of NM-92 cultivar of a Pakistani green gram.MATERIALS AND METHODSThe present study was replicated thrice in the plant laboratory of Department of Genetics, Faculty of Science, and University of Karachi. The seeds of mung bean cultivar NM-92 were acquired from National Agricultural Research Centre (NARC), Islamabad. These freshly collected 15 seedsˉ1 treatment / replication were divided into two sets. The first was named as sodium chloride (SC) stress treatments were imbibed in sterile distilled water (DW) whereas, second set soaked in gibberellic acid (GAᴣ) (BDH Chemicals, England), indole acetic acid (IAA) (Fluka, Switzerland), and salicylic acid (SA) (J.T. Baker, Holland) in the separate beaker for 24 hours under dark condition. After 24 hours, given ample time to both the sets at room temperature. After recovery, all 20 treatments were sown in the 150 X 30 mm sized petri-dishes containing 0, 50, 150, 250, and 350 millimolar (mM) sodium chloride solution (Fisher Scientific, UK) for 72 hours.Protein extraction: Protein extraction was done by taking 0.3g of seedlings in an ice chilled mortar and crushed by adding 600µL 0.2 M Tris-HCl buffer having pH 7.5 contained 5% SDS (w/v) and 5% 2-mercaptoethanol (v/v). The homogenate was incubated at 0oC for 30 min., boiled in the water bath for 3 min. at 100oC. Samples were centrifuged in Heraeus Biofuge D-37520, Germany for 30 min. at 8000 rpm. The protein supernatant was saved at below 0°C for quantitative and qualitative determination with minor modifications. The total soluble protein content of the samples was estimated via “Bovine Serum Albumin (BSA) standard curve” and explicit in µg protein milligramˉ1 fresh weight of mung seedlings.Bovine serum albumin standard curve (2000 μg/mL): Total protein standard curve was made by dissolving 0.05g of Bovine Serum Albumin (BSA) in 25mL of distilled water. Ten serial dilutions were made from 0.1 mL to 1mL by BSA solution then performed Lowry. A standard curve of total proteins was plotted by taking BSA absorbance at Y-axis and 2000 μg BSA / mL at X-axisSample preparation for SDS-PAGE: For qualitative assessment of total proteins; the 35μL of saved protein supernatant was combined with 15μL of sample diluting buffer (SDB). The SDB was made up of 0.0625 M Tris-HCl pH 6.8 with 2% of SDS, 10% of glycerol, 0.003% of bromophenol blue dye and 5% of 2-mercaptoethanol. Boil the 50μL protein SDB supernatant at 100oC in water bath for 3 min., centrifuged at 6000 rpm for 4 min. The supernatant was loaded on SDS-PAGE gel with the given formulae. The SDS- PAGE: Total proteins were fractionated via SDS-PAGE with 4% stacking and 12% resolving gel. The resolving gel of 12% was made by taking 6mL solution A, 1.8 mL 3 M Tris 1 M HCl buffer pH 8.8, 144μL 10% SDS, 5.74 mL sterile distilled water, 720μL 1.5% ammonium persulphate (APS) in deionized water and 10μL TEMED. While, stacking was composed of 1.25mL of solution A, 2.5mL of 0.5M Tris 1M HCl buffer pH 6.8, 100μL 10% SDS, 1.8 mL of distilled water, 500μL 1.5% APS and 12μL TEMED. Solution A was prepared by conjoining 30% acrylamide and 0.8% N, N’-methylene-bisacrylamide in deionized water. To avoid polymerization in the beaker; the prepared solution was quickly poured into the 3 mm thick gel plates after adding TEMED. The stacking was lined over resolving gel, then combs were inserted between the gel plates of SCIE-PLAS TV-100 separation system, UK, and allowed to polymerize for ½ an hour. After polymerization gel was placed in the tank which were filled with Tris-Glycine buffer (electrode buffer) pH 8.4 then combs were removed. The electrode buffer contained 0.3% Tris, 1.41% Glycine and 0.1% SDS in 2000mL d/w. The gel was pre-run for 15 min. at 60 volts and 120 mA currents. The prepared SDS-PAGE samples were loaded in wells with BlueStepTM Broad Range Protein Marker, AMRESCO, USA as standard and run at 60 volts & 120 mA for about 45 min. When samples entered in resolving gel, and then gave 100 volts and 200 mA currents for around 2.5 hours. Furthermore, electrophoresis was carried out at a constant watt.The Gel was washed with 30% ethanol on Uni Thermo Shaker NTS-1300 EYELA, Japan at the constant shaking for 30 min. Then gels were placed in 10% glacial acetic acid in 50% methanol solution (Fixative) for 24 hours. SDS Gel was stained until protein bands were visible thereat placed as 5% of Methanol in 7.5% acetic acid glacial solution to destain the bands background. SDS-PAGE stain composed of 0.125% coomassie brilliant blue R-250 dissolved in 40% of Methanol and 7% acetic acid glacial solution. The stain was stirred on Magnetic stirrer & hot plate M6/1, Germany for 6-10 hours before used. Photographs were taken by Sanyo digital camera VPC-T1284BL and bands were scored through numbering pattern. Gels preserved in 10% acetic acid solution at 4°C.Interpretation of bands and data analysis: The total soluble protein bands relative mobility calculated by below formulae and Dendrogram was constructed via SPSS v. 20Where,F=(Migrated distance of protein band)/(Migrated distance of dye front)Slop=(Log MW of protein marker lower limit band–log〖MW of protein marker upper limit band )/(RF protein marker lower limit band –RF of protein marker upper limit band)RESULTS:The total soluble proteins extracted from green gram were perceived by SDS-PAGE Blue StepTm broad range biochemical markers. The protein-based marker was used to evaluate the toxic effect of sodium chloride along with pre-treatments of GAᴣ, IAA, and SA on proteome assay. In the current work, seedlings total soluble proteome resolved 24 polypeptide bands ranging from 200 to 17.1 kDa were recognized by using SDS-PAGE. The figure 1 showed Dendrogram assay, which classified the 20 treatments of SC, GAᴣ, IAA and SA into two major clusters where, the cluster I was the largest one (figure 1). Cluster I consisted of 15 treatments that further divided into I-A, and I-B. The pre-treatments of SC50+SA, SC150+SA, SC250+SA, and SC350+IAA were grouped together into C-1 of sub-cluster I-A. The C-2 of sub-cluster I-A, pre-treatment SC350+SA was most diverse among 20 treatments. The C-1 treatments showed 99% homology when compared with each other while, it was 97% similar with C-2. The sub-cluster I-B comprised another 10 treatments, SC0+GAᴣ, SC50+GAᴣ, SC150+GAᴣ, SC250+GAᴣ, SC350+GAᴣ, SC0+IAA, SC50+IAA, SC150+IAA, SC250+IAA, and SC0+SA that were also 99% similar for total proteins. Sub-cluster I-B pre-treatments was exhibiting 94% homology with the sub-cluster I-A. The second cluster was the smallest one that was divided into two sub-clusters, II-A and II-B. The II-A was comprised of SC50, SC150, and SC250 while, sub-cluster II-B consisted of SC0 and SC350. Within each sub-cluster, pre-treatments expressed 99% homology whereas, II-A was 97 different from II-B. Furthermore, cluster I showed 75% similarities with cluster II (figure 1). The seedlings storage proteome profile of green gram was shown in table 1.The results showed that 120kDa, 114.6 kDa, 51.8 kDa, 29.1 kDa and 22.8 kDa proteins bands were not induced at 0 mM SC, GAᴣ, IAA, and SA. The table 1 depicted the presence of 120 kDa and 114.6 kDa bands only at 350 mM SC level with all phytohormones treatments. Similarly, 51.8 kDa protein bands were appearing at 150SC, 250SC and 350SC stress with phytohormones. Based on the information collected from the NCBI protein database, this peptide was related to the 8S globulin alpha subunits. The two other, 7S globulins sub-units having 29.1kDa and 22.8 kDa molecular weights bands were synthesized under 50mM, 150mM, 250mM, 350mM SC stress with phytohormones. Concerning protein polypeptide of molecular weight 104.5 kDa, 99.8 kDa, 91.0 kDa, 55.0 kDa, and 46.0 kDa, those were induced by GAᴣ, IAA and SA at 0 to 350 mM SC. While, 17kDa protein band was appearing in SA, and IAA treated samples and 95.3kDa band was only present in SA treatment. Other 12 protein bands were present in all treatments proved as house-keeping proteins of green gram (table 1).DISCUSSIONThe SDS-PAGE profiling for proteome is the reliable and applied biochemical approach that has been used as biochemical marker in various crop differentiation, and characterization. In the current study, first time SDS-PAGE was utilized to investigate the impact of GAᴣ, IAA, and SA pre-soaking on green gram under salt toxicity. The salt toxicity adversely affects all seed, seedling, and plant metabolic process (Parveen et al., 2016). At salt toxicity, the endogenous GAᴣ, IAA, and SA levels markedly decrease (El-Khallal et al., 2009). In such condition, exogenous application of GAᴣ, IAA, and SA enhance seedlings survival rate by increasing synthesis of seed storage proteins. Likewise, our Dendrogram characterization based on 20 treatments showed significant diversity under 0 to 350 mM SC stress. The salicylic acid treatments were grouped together except SC0+SA treatment, exhibiting a close relationship, which proved its acclimating role under salt stress. These findings will help plant breeder toward enhancing food quality and quantity of green gram in future breeding programme on saline sodic land.The SDS-PAGE assay revealed 200. kDa, 109.4 kDa, 77 kDa, 68 kDa, 49 kDa, 38 kDa, 33 kDa, 26 kDa, 24 kDa, 22 kDa, 21 kDa and 19 kDa fractions as essential green gram proteins. Among these, 68 kDa, 49 kDa, 33 kDa, 26 kDa, 24 kDa and 21 kDa peptides were seed biotinylated isoform protein (Riascos et al., 2009), putative NADH-ubiquinone oxidoreductase subunit H (Gostinčar et al., 2019), heat shock protein 33 (Hamidian et al., 2015), globulin protein, seed coat / maturation protein (Dhaubhadel et al., 2005), and protein for dimerization. While, 22 kDa proteins belonged to the class of prolamin alpha zein Z1C1_2, Z1C1_4, and Z1C1_8 precursors, and 19kDa peptide was related with Z1A1_2, Z1A2_2, and Z1B_6 precursors (Miclaus et al., 2011). Further, the 91 kDa peptide is sucrose synthase SS1 protein, and 77kDa protein is the NADPH-cytochrome P450 reductase (Wang et al., 2004). Also, the phosphatase-associated two other proteins having 46 and 55 kDa molecular weight were reported earlier in Mucuna pruriens. Hameed et al. (2012) and Malviya et al. (2008) found 55 and 46kDa peptides as 7S vicilin small sub-units and 17kDa as 11S globulins sub-unit in the studied Vigna radiata. Some other molecular weight proteome such as 68 kDa and 49kDa are 7S vicilin, 33kDa is 8S vicilin, 38 and 26kDa 8S globulins, 24kDa 11S globulins, and 22kDa 16.5S globulins. These proteins required for germination and seed establishment of green gram plant (Hameed et al., 2012).The vast accumulation of 23kDa and 22kDa peptides under salt stress by salicylic acid, were reported previously in the mangrove Bruguiera parviffora and Zea mays (El-Khallal et al., 2009). Correspondingly, El-Kafafi et al. (2015) reported the presence of 115kDa, 23kDa, and 22kDa bands in the salt tolerant lines of green gram. These proteomes induced under salt stress may play a pivotal part in the stress acclimation and osmotic adjustment. Similarly, the induction of 104 kDa and 100kDa MW polypeptide by SC stress in the salt tolerant genotypes of green gram indicated the functional role of phytohormones in various metabolic and defense response El-Kafafi et al. (2015); Alharby et al. (2016), El-Khallal et al. (2009), Qados (2010). Ali et al. (2007), Alharby et al. (2016), and El-Kafafi et al. (2015) observed 17kDa, 26kDa, 33kDa and 77kDa bands involving in salt tolerance and can be considered as a positive biochemical marker for salt stress. Further, 26 kDa MW peptide also functions as osmotin under the salt stress that involved in enhancing the accumulation of glycine betaine and proline in the cells. Hence, proteome assay of green gram showed that GAᴣ, IAA, and SA could regulate the expression of salt stress proteins that are anticipated to play a crucial part in the salt tolerance mechanism. Likewise, the involvement of phytohormones in the induction of changes in the proteome profile pattern was attributed to their part in managing cell division by regulating some genes of apical meristems.CONCLUSIONFinally, the results revealed the presence of the ten new bands with MW of 200kDa, 120 kDa, 114.6 kDa, 109.4kDa, 104.5kDa, 99.8kDa, 95.3kDa, 51.8kDa, 29.1kDa and 22.8kDa have not reported previously under salt stress with phytohormones treatments in green gram. Furthermore, it was observed that phytohormones alleviate the negative impact of salt stress on green gram by enhancing synthesis of salt defense polypeptides. Hence, higher accumulation of proteins was observed in salicylic acid treated seedlings. Thus, present work recommended the pre-soaking of phytohormones to overcome the toxic impact of sodium chloride on green gram. 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