Academic literature on the topic 'Blueprint A 3.0'

Gene expression in early stage embryos relies mostly on post-transcriptional control of maternal transcripts accumulated during oocyte maturation. However, while the building process to obtain a competent oocyte is now better understood, it is still not clear what transcriptome blueprint composes a competent oocyte. The aim of the study was to compare the mRNA expression pattern between oocytes collected from fertile heifers and repeat breeders by using RNAseq. Oocytes were collected by ovum pickup from 3 heifers that were 11–15 months of age and became pregnant at the following oestrus and from 4 adult cows with an age of 4 to 7 years, classified as repeat breeders after they failed to become pregnant for a minimum of 3 consecutive AI. To obtain oocytes from follicles with the same degree of development, at time 0 all follicles visible through transrectal ultrasound examination were removed by transvaginal aspiration. Five days later oocytes were collected by ovum pick up from the newly formed follicles with diameters >5 mm. Oocytes from each animal were pooled and sequenced as a single sample. Total RNA was extracted by RNeasy Micro Kit (Qiagen, Valencia, CA, USA). Amplified cDNA, was prepared starting from total RNA using the Ovation RNA-Seq System V2 (Nugen Technologies, San Carlos, CA, USA). After library preparation with TruSeq DNA Sample Prep kit (Illumina, Madison, WI, USA), sequencing was performed on an Illumina HiSEqn 2000. Galaxy and Chipster open web-based platforms were used to analyse the data. We identified 49 differentially expressed genes. Heifers’ oocytes mRNA pattern indicated greater potential to sustain cell division. In particular, oocytes expressed more Keratin 14 (a gene involved in cell proliferation) and kinesin family member 20B (a protein involved in cytokinesis). More competent oocytes also have a greater ability to repair single-strand breaks due to the high levels of endo/exonuclease (5′-3′), endonuclease G-like. This may reflect greater capacity to neutralise DNA damage and, therefore, greater ability to preserve and transmit high-quality DNA. Repeat breeders portray a different landscape; their greater expression of Jun oncogene, Heat shock protein 1, Stimulated by retinoic acid gene6, arylhydrocarbon receptor nuclear translocator, fibromodulin, and aryl-hydrocarbon receptor repressor suggest that these oocytes have been subjected to environmental stress during oocyte maturation. Their greater expressions of inhibin α, stearoyl-CoA desaturase, junctional adhesion molecule 2 have been previously shown to correlate with a reduced oocyte developmental potential. Furthermore, the Cannabinoid receptor protein 1 expression suggests a compromised ion function that can lead to a failed activation of the development program. Finally, the greater expression of Ubiquilin3 and Heat shock protein 1 led to high protein and mRNA degradation, respectively, suggesting that these oocytes are deprived of essential components to sustain embryo growth. In conclusion our data provide the first detailed snapshot of the mRNA pattern defining the differences between a competent oocyte and an incompetent oocyte in vivo. Study supported by PRIN 2008, 2009 and EU-Quantomics.

91 Background: Programmed cell death ligand 1 (PD-L1) expression by immunohistochemistry (IHC) enriches for responses to PD-1/PD-L1 inhibitors, however its role as a predictive biomarker in hepatocellular carcinoma (HCC) is inconclusive, with no consensus on any particular assay. We evaluated the performance of 4 different PD-L1 detection assays previously published in landmark clinical studies. Methods: PD-L1 IHC was performed on 4 serial sections from tissue microarray (TMA) blocks containing 100 archival cases of HCC that included tumour and surrounding non-tumorous tissue. Antibody clones E1LN3, 28-8, 22c3, SP263 were compared on the basis of percentage and intensity of staining in malignant cells (M) to generate an H-score (range 0-300). Immune cells infiltrating (ICI) and at the periphery, in non-tumorous tissue (ICP) were scored on a 4-tier system (0-3). Results: Patients were 76% males, 20% HCV-positive, 64% cirrhotic with a median age of 67 years. Median tumour size was 4 cm, 70% of patients had T1-T2 tumours and 48% were of grade 2. The proportion of PD-L1 positive cases according to M-ICI-ICP pattern was 2-6-2% for E1LN3, 10-18-19% for 28-8, 9-22-18% for 22c3 and 5-14-13% for SP263. Pairwise comparison of M H-scores revealed heterogeneity across antibodies, with highest concordance between E1L3N/SP263 (R2 = 0.95), E1L3N/22c3 (R2 = 0.65), 22c3/SP263 (R2 = 0.66) and increasing discordance for 28-8/22c3 (R2 = 0.44), E1L3N/28-8 (R2 = 0.29), and 28-8/SP263 (R2 = 0.26). Detection of PD-L1-positive immune infiltrates using a semi-quantitative scoring system revealed significantly different scores in pairwise non-parametric comparisons of ICI (p < 0.05) but not ICP (p > 0.05 for chi-square test). Conclusions: In the Blueprint-HCC study we demonstrated that quantification of PD-L1 protein levels in tumour cells, intra-tumoural and peri-tumoural infiltrate is characterised by inter-assay discordance in HCC. This has profound implications in the clinical development of predictive correlates of efficacy to immunotherapy in HCC. Sources of such discordance should be explored.

STEM Education through the Malaysia Education Blueprint 2013–2025 (PPPM 2013–2025) is an important agenda in the transformation of education to prepare the younger generation for the challenges of the 21st century. Over the years, STEM was carried out, but there are still some issues which contribute towards the failure in achieving a policy percentage set of 60% science and 40% literary studies in secondary schools. The target to increase the number of Science students was not achieved. Therefore, this study was conducted to produce a STEM@IDEAS module as an alternative to increase students’ interest and understanding in solving the synopsis of learning in science, technology and mathematics (STEM). The module STEM@IDEAS focuses in competition design and generating prototype products through a variety of synopsis statements. STEM practices provide students with various trainings such as application of knowledge, skills and assessment to solve synopsis. This study has five (5) STEM practice steps and a total of three (3) modules which will be applied using the STEM elements. Furthermore, the STEM@IDEAS module was tested on several groups of four secondary school students around Pasir Gudang, Johor. A questionnaire was used to evaluate if STEM@IDEAS modules are in line with STEM and its impact on students. This survey uses a Likert scale of 0 to 4 to evaluate starting from 0 (strongly disagree) up to 4 (strongly agree) for each question submitted in each section. The STEM@IDEAS module and the above study are expected to be a source of interest and an alternative way for schools to support the nation’s education policy in strengthening the education development plans towards the nation’s progress.

Introduction: Advanced systemic mastocytosis (AdvSM) comprises a heterogeneous group of clonal mast cell neoplasms, primarily driven by KIT D816V. Measures of AdvSM response, including the International Working-Group for Myeloproliferative Neoplasms Research and Treatment and European Competence Network on Mastocytosis (IWG-MRT-ECNM) criteria, are based on improvements in mast cell-related organ damage (C-findings), and further sub-classified by the extent of reduction in measures of mast cell disease (e.g. serum tryptase level, bone marrow mast cell burden). However, assessment of some C-findings lacks precision (such as splenomegaly and its resolution). Normalization of C-findings may not be an adequate surrogate for important clinical outcomes such as overall survival (OS). We evaluated whether pure pathologic response (PPR) criteria based on changes in bone marrow mast cells, serum tryptase, and complete blood count was more closely correlated with OS compared to the modified IWG-MRT-ECNM (mIWG-MRT-ECNM) criteria. Methods: As an exploratory post-hoc analysis of the phase 1 EXPLORER study of avapritinib in AdvSM, we evaluated responses lasting ≥12 weeks by both mIWG-MRT-ECNM and PPR criteria. At baseline, evaluability for mIWG-MRT-ECNM response required ≥1 evaluable C-findings; PPR required presence of bone marrow mast cell aggregates and/or serum tryptase ≥20 ng/mL. Per PPR, morphologic complete remission (mCR) is absence of bone marrow mast cell aggregates, serum tryptase <20 ng/mL and full (or partial [mCRh]) hematologic recovery; morphologic partial remission (mPR) is ≥50% reduction in bone marrow mast cells and serum tryptase level. OS was analyzed by Kaplan-Meier method and was time from first dose to death. OS comparisons were by log-rank test, performed for best response and landmark analyses at various cycles. Results: As of the data cut-off of August 30, 2019, 80 patients enrolled including 62 with AdvSM (7 with aggressive SM [ASM], 44 SM with an associated hematologic neoplasm [SM-AHN] and 11 with mast cell leukemia [MCL]). Ten (16%) AdvSM patients (7 ASM, 3 SM-AHN) were not response evaluable (RE) per mIWG-MRT-ECNM criteria, due to a lack of an evaluable C-finding at baseline, and 4 additional AdvSM patients were recently enrolled and were not yet response evaluable. Of the 48 RE patients (3 ASM, 35 SM-AHN and 10 MCL) the best overall response rate (ORR) per mIWG-MRT-ECNM was 77% (8% CR, 19% CRh, 42% partial response [PR], and 8% clinical improvement [CI]). Non-responders had stable disease (SD; 21%) or were not evaluable (NE) due to insufficient (<13 weeks) follow-up (2%). Responders (CR/CRh/PR/CI) had 18-month OS of 85% (CR/CRh, 100%; PR, 77%; CI, 75%; Figure 1A); non-responders had 18-month OS of 48% (SD, 53%; NE, 0%) (P=0.042). Per PPR criteria, the best ORR was similar at 79%; however, a greater proportion of patients were assessed as being in a complete remission (15% mCR, 27% mCRh and 38% mPR). Non-responders by PPR all had SD (21%). This demonstrates that elimination of measurable mast cell burden can be discordant with complete C-finding resolution. Responders (mCR/mCRh/mPR) by PPR had 18-month OS of 88% (mCR/mCRh: 100%; mPR: 72%; Figure 1B); non-responders (all SD) had 18-month OS of 21% (P=0.0001). Eventually, all 62 AdvSM patients will be evaluable by PPR criteria, including those 10 patients without mIWG evaluable C-findings at baseline; however, 5 patients had insufficient follow-up at the time of analysis. For the 57 AdvSM patients with sufficient follow up, the best ORR per PPR criteria was similar at 77% (14% mCR, 26% mCRh and 37% mPR). Overall, no patients had a best response of progressive disease based on mIWG or PPR criteria. Landmark analyses of PPR at the end of 6 cycles showed a trend in 18-month OS of mCR/mCRh>mPR>SD in patients with similar starting avapritinib doses of ≥200 mg daily (n=48 of 57 PPR-evaluable patients). Conclusions : In the phase I EXPLORER study, response assessment in AdvSM using PPR criteria increases the evaluable population, significantly correlates with OS, and should be explored as a potential primary endpoint for future trials. Disclosures Gotlib: Blueprint Medicines Corporation: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Chair of the Response Adjudication Committee and Research Funding, Research Funding; Deciphera: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: co-chair of the Study Steering Committee and Research Funding. Radia:Blueprint Medicines Corporation: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees, Other: Education events. George:Blueprint Medicines Corporation: Consultancy, Other: I have received no funding for this research. ARUP Laboratories, owned by the University of Utah, has received funding; Allakos: Consultancy; Deciphera: Other: consultancy, but has received no financial compensation for the past 12 months; Celgene: Consultancy. Robinson:Blueprint Medicines Corporation: Research Funding. Drummond:Jazz: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Gilead: Membership on an entity's Board of Directors or advisory committees; Bristol Myers Squibb: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Takeda: Membership on an entity's Board of Directors or advisory committees; Astellas: Membership on an entity's Board of Directors or advisory committees; Blueprint Medicine Corporation: Research Funding. Bose:Incyte Corporation: Consultancy, Honoraria, Research Funding, Speakers Bureau; Kartos Therapeutics: Honoraria, Research Funding; Astellas Pharmaceuticals: Research Funding; NS Pharma: Research Funding; Pfizer, Inc.: Research Funding; Promedior, Inc.: Research Funding; Constellation Pharmaceuticals: Research Funding; Celgene Corporation: Honoraria, Research Funding; CTI BioPharma: Honoraria, Research Funding; Blueprint Medicines Corporation: Honoraria, Research Funding. Hexner:Samus Therapeutics: Research Funding; Novartis: Research Funding; American Board of Internal Medicine: Other: member of the hematology exam committee; Blueprint Medicines Corporation: Other: serves on a data safety monitoring committee, Research Funding. Winton:Blueprint Medicines Corporation: Research Funding; Samus Therapeutics: Research Funding; Incyte Corporation: Research Funding. Horny:Novartis: Consultancy; Blueprint Medicines Corporation: Consultancy. Tugnait:Blueprint Medicines Corporation: Current equity holder in publicly-traded company, Ended employment in the past 24 months. Schmidt-Kittler:Blueprint Medicines Corporation: Current equity holder in publicly-traded company, Ended employment in the past 24 months. Evans:Blueprint Medicines Corporation: Current equity holder in publicly-traded company, Ended employment in the past 24 months. Lin:Blueprint Medicines Corporation: Current Employment, Current equity holder in publicly-traded company. Mar:Blueprint Medicines Corporation: Current Employment, Current equity holder in publicly-traded company. Deininger:Leukemia & Lymphoma Society: Research Funding; Bristol-Myers Squibb: Consultancy, Honoraria, Other, Research Funding; Ariad: Consultancy, Honoraria, Other; Incyte: Consultancy, Honoraria, Other, Research Funding; Novartis: Consultancy, Other, Research Funding; Medscape: Consultancy; Takeda: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Fusion Pharma: Consultancy; Blueprint Medicines Corporation: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: part of a study management committee, Research Funding; DisperSol: Consultancy; Galena: Consultancy, Honoraria, Other; Celgene: Research Funding; Gilead Sciences: Research Funding; SPARC: Research Funding; Sangamo: Consultancy, Membership on an entity's Board of Directors or advisory committees; Pfizer: Honoraria, Other, Research Funding. DeAngelo:Incyte Corporation: Consultancy; Glycomimetics: Research Funding; Forty-Seven: Consultancy; Amgen: Consultancy; Novartis: Consultancy, Research Funding; Pfizer: Consultancy; Abbvie: Research Funding; Jazz: Consultancy; Autolos: Consultancy; Shire: Consultancy; Takeda: Consultancy; Blueprint Medicines Corporation: Consultancy, Research Funding; Agios: Consultancy.

AbstractEighteen purebred steers of three genotypes, Aberdeen Angus (AA), Charolais (CH) and Holstein (HO), were divided within genotype into three groups of six animals and offered one of three different levels of feeding either moderate (M/M) or high (H/H) both for 20 weeks or moderate for the first 10 weeks followed by high for the remaining 10 weeks (M/H). Growth rates during the final 10 weeks of the experimental period differed between dietary regimen (M/M = 0·87; M/H = 1·25; and H/H = 1·02 kg/day; s.e.d. = 0·08;P< 0·001). Over the entire 20 week experimental period animals offered the M/M level of feeding grew more slowly (0·97 kg/day) than those offered the M/H and H/H level of feeding (1·20 kg/day; s.e.d. = 0·06;P< 0·001). Mean growth rates for CH, HO and AA steers were 1·21, 1·13 and 1·03 kg/day (s.e.d. = 0·06;P< 0·05). The animals were all slaughtered at a fixed age of 18 months, according to the Meat and Livestock Commission Blueprint for beef and, 48 h post mortem, samples of m. longissimus lumborum (LL) and m. vastus lateralis (VL) were removed for analyses.Muscle fibres were classified histochemically, according to their contractile and metabolic properties, and muscle fibre size was measured. Fibre type frequency was calculated and, in LL, the total fibre number of the muscle was estimated. There was little impact of feeding level, or consequentially growth rate, on muscle fibre frequency and size. The effects seen were confined mainly to LL where there were significant differences between the M/M and H/ H groups with respect to fast twitch glycolytic fibres (mean % frequency (M/M = 40·1 and H/H = 44·3; s.e.d. = 1·4;P< 0·01); mean % area (M/M = 51·9 and H/H 56·0; s.e.d. = 1·5;P< 0·05)) and apparent total fibre number (M/ M = 35·0; and H/H = 41·9 ✕ 104; s.e.d. = 1·7;P< 0·05) which were greater in H/H than in M/M groups. However, in both LL and VL the predominant differences were related to genotype; in particular, overall fibre size was smallest in CH, while slow oxidative (SO; type I) fibre area was highest in AA. For LL, analysis across all animals showed a positive relationship between SO area, % area, % frequency and overall acceptability of meat at 14 days as evaluated by a trained sensory panel. No such relationship was observed for VL. The data suggest that in this study manipulation of feeding level has only a small impact on muscle fibre characteristics and that the differences between genotype and muscle type may be more important in determining the variability of overall acceptability than growth rate.

Abstract Introduction: The KIT D816V oncogene is a key driver in 90-95% of patients with systemic mastocytosis (SM), a group of mast cell (MC) neoplasms including indolent SM (ISM), smoldering SM (SSM) and AdvSM. Debilitating symptoms related to MC proliferation and degranulation characterize ISM and SSM and are also prominent in AdvSM, which is further complicated by SM-related organ damage and decreased survival. Currently, there are no approved agents that selectively target KIT D816V, and there are limited tools to assess symptom improvement in SM. Avapritinib, a highly potent and selective inhibitor of the KIT D816V mutant, showed substantial clinical activity in AdvSM (83% overall response rate (ORR) per modified IWG-MRT-ECNM criteria) in the Phase 1 EXPLORER study [Deininger, et al, EHA, 2018]. We present results using a novel PRO questionnaire, the AdvSM-SAF, developed in accordance with FDA guidance, to assess changes in symptoms in the Part 2 dose expansion phase of EXPLORER. Methods: Patients (pts) received avapritinib at the recommended Phase 2 dose (300 mg once daily [QD]) in continuous 28-day (d) cycles. Pts completed the AdvSM-SAF and 2 additional PROs used in other cancers, the Patient Global Impression of Symptom Severity (PGIS) and the European Organization for Research and Treatment of Cancer Quality of Life (QLQ). PROs and KIT D816V mutant allele fraction (MAF) in blood were assessed serially as follows: AdvSM-SAF daily, from 7 d before first avapritinib dose, ie, baseline (BL: Cycle [C]1 Day [D]1) and through C12, using an electronic diary; PGIS and QLQ at BL (C1D1) and on D1 of each cycle through C12; and KIT D816V mutant allele fraction (MAF) at BL and on D1 of C3, 7, 11, and then every 6 cycles. The AdvSM-SAF assesses severity of 8 symptoms (pruritus, flushing, spots, nausea, vomiting, diarrhea, abdominal pain, fatigue) on a 0-10 scale, and 2 items assess frequency of diarrhea and vomiting. Results are analyzed as a Total Symptom Score (TSS), combining all 8 severity items (maximum symptom score=80), and as a GI domain (combining 4 symptoms: nausea, vomiting, diarrhea, abdominal pain; maximum score 40) and Skin domain (combining 3 symptoms: pruritus, flushing, spots; maximum score 30). Analyses were based on 7 d average scores. AdvSM-SAF data were summarized at BL (C1D-7 to C1D-1), and over the 7-d interval prior to C3D1 and C7D1, and correlated with QLQ, PGIS and KIT D816V MAF. Results: As of 22 June 2018, 25 pts were treated in Part 2 of the study; 24 are ongoing, and enrollment and follow-up continues. The median duration of avapritinib treatment was 5.7 mo (range, 1.7+ to 10.3+ mo). AdvSM-SAF scores for 24 pts with data are shown the Table. Mean BL TSS, GI and Skin scores were 22.6, 9.1, and 7.1. The most severe BL symptom was fatigue (mean score of 6.4). Among 21 pts with scores at C3D1 and 12 pts with scores at C7D1, mean reductions from BL TSS, GI and Skin scores were 6.4, 3.9, and 1.5 points, and 11.1, 5.7, and 4.4 points, respectively, indicating further improvement with continued treatment. Symptom reductions were seen for all 8 items at C3D1. Further symptom improvement was observed for 6 of 8 items from C3D1 to C7D1. Reductions in AdvSM-SAF TSS, GI and Skin scores significantly correlated with improvement in QLQ Emotional (EF) and Cognitive functioning (CF) scales (p values for Pearson Correlation Coefficients <0.0001, <0.0001, and 0.04 with EF and <0.0001, 0.0001, and 0.04 with CF, respectively). Among 18 pts with BL PGIS (mean 3.2, SD 1.15, range 1-5), improvement in PGIS was highly associated with improvement in GI score (p value 0.0004). Decreases in KIT D816V MAF correlated with reduction in TSS (p value 0.04). Conclusions: Avapritinib treatment resulted in meaningful symptom improvement as measured by the AdvSM-SAF in overall symptoms (TSS), GI and skin domains, and all individual symptoms, and improvement continued with longer duration of treatment. Reduction in AdvSM-SAF scores correlated with improvements in other PRO instruments, PGIS and QLQ, supporting the AdvSM-SAF as a useful new tool to assess symptoms in pts with AdvSM. Reduction in AdvSM-SAF scores also correlated with decreases in KIT D816V MAF, indicating that reductions in disease burden may correlate with reduced disease symptoms and improved quality of life. These data warrant further development of avapritinib in AdvSM, as well as in ISM and SSM where symptom burden and poor quality of life are the predominant disease manifestations. Disclosures Gotlib: Deciphera: Consultancy, Honoraria, Research Funding; Blueprint Medicines: Consultancy, Honoraria, Research Funding; Novartis: Consultancy, Honoraria, Research Funding; Promedior: Research Funding; Celgene: Consultancy, Honoraria, Research Funding; Kartos: Consultancy; Incyte: Consultancy, Honoraria, Research Funding; Gilead: Consultancy, Research Funding. Radia:Novartis: Speakers Bureau; Blueprint: Consultancy. DeAngelo:Glycomimetics: Research Funding; Shire: Honoraria; Pfizer Inc: Consultancy, Honoraria; Incyte: Consultancy, Honoraria; Amgen: Consultancy; Novartis Pharmaceuticals Corporation: Consultancy, Honoraria; Takeda: Honoraria; ARIAD: Consultancy, Research Funding; BMS: Consultancy; Blueprint Medicines: Honoraria, Research Funding. Bose:CTI BioPharma: Research Funding; Celgene Corporation: Honoraria, Research Funding; Pfizer, Inc.: Research Funding; Blueprint Medicines Corporation: Research Funding; Incyte Corporation: Honoraria, Research Funding; Astellas Pharmaceuticals: Research Funding; Constellation Pharmaceuticals: Research Funding. Conlan:Blueprint Medicines: Employment. Oren:Blueprint Medicines: Employment. Shi:Blueprint Medicines: Employment. Deininger:Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Blueprint: Consultancy.