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Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm characterized by a reciprocal translocation between the long arms of chromosomes 9 and 22 that results in expression of the oncoprotein BCR-ABL1. An optimal response to tyrosine kinase inhibitors (TKIs) requires a BCR-ABL transcript level ≤ 10% at 3 months, ≤ 1% at 6 months, ≤ 0.1% at 1 year, and ≤ 0.01% onwards. Complex scenarios like P190 BCR-ABL CML, unusual BCR-ABL transcripts, primary refractory CML, and detection of TKI-resistance mutations during treatment frequently pose a therapeutic challenge. In this article we present some of these clinical scenarios using a case-based approach. J Med Cases. 2020;11(7):215-220 doi: https://doi.org/10.14740/jmc3510

Injury to any individual structure in the knee interrupts the overall function of the joint and initiates a cascade of biological and biomechanical changes whose endpoint is often osteoarthritis (OA). The knee meniscus is an integral component of knee biomechanics and may also contribute to the biological homeostasis of the joint. Meniscus injury altering knee function is associated with a high risk of OA progression, and may also be involved in the initiation of OA. As the relationship between meniscus injury and OA is very complex; despite the availability of transcript level data on human meniscus injury and meniscus mediated OA, mechanistic studies are lacking, and available human data are difficult to validate in the absence of patient-matched noninjured control tissues. As similarities exist between human and mouse knee joint structure and function, investigators have begun to use cutting-edge genetic and genomic tools to examine the usefulness of the mouse as a model to study the intricate relationship between meniscus injury and OA. In this review, we use evidence from human meniscus research to identify critical barriers hampering our understanding of meniscus injury induced OA and discuss strategies to overcome these barriers, including those that can be examined in a mouse model of injury-mediated OA.

Coronary artery bypass grafting (CABG), together with percutaneous coronary intervention (PCI), are both still the most efficient procedures for myocardial revascularization to treat advanced coronary artery disease (CAD). Donor blood vessels used in CABG are usually the internal thoracic artery (ITA) and saphenous vein (SV). The importance of inflammation and inflammatory pathways in graft patency is well established. Nevertheless, not all molecular mechanisms underlying the inflammatory process appear to be clear. Employing the expressive microarray approach to analyze the transcriptome of both venous and arterial grafts, five GO BP terms has been selected: “cellular response to interferon-gamma”, “inflammatory response”, “interferon-gamma-mediated signaling pathway”, “response to interferon-gamma” and “positive regulation of inflammatory response”. This study aimed to evaluate potential molecular factors that could be characteristic markers for both SV and ITA conduits. Running title: Aortocoronary conduits may show a different inflammatory response

Purpose: The early BCR-ABL1 reduction had the prognostic impact of the chronic-phase chronic myeloid leukemia (CML-CP) patients. This study was to find a more precise early prognosis index at 3 months in the patients with newly diagnosed CML-CP, especially for the patients with BCR-ABL1IS >10%. Methods: We retrospectively analyzed the data of 79 newly diagnosed CML-CP patients from October 2013 to April 2017. All patients took imatinib regularly and continuously and monitored BCR-ABL1 transcript level at baseline and 3, 6, 9, 12, 18 months after starting imatinib treatment. Results: Among the 44(55.7%) patients with BCR/ABL1IS ≤10% at 3 months after imatinib treatment, 12(27.3%) cases did not achieve major molecular response (MMR) at 12 months, and 7(14.9%) patients with the halving time BCR-ABL1 transcript ≤40 days failed to achieve MMR at 12 months. However, approximately twenty-six percent of the patients with BCR-ABL1IS >10% still obtained MMR. Moreover, the patients with BCR-ABL1IS ≤10% and halving time ≤40 days had a significantly better MMR than that of the patients with the BCR-ABL1IS ≤10% and halving time >40 days (88.6% versus 11.1%, P 10% and halving time >40 days rarely achieved MMR at 12 months. Conclusion: These data indicated that the halving time of BCR-ABL1 transcript was also an important prognostic factor as that of the BCR-ABL1IS. Combined observations of these two prognosis indexes are more accurate predictor for the long-term molecular response, especially for the CML-CP patients with BCR-ABL1IS >10%, and which is helpful for TKI switching as early as possible to improve patients' survival and reduce drug costs.

Drug sensitivity biomarkers are molecular features informative for drug response. Previous studies have identified many candidate drug-sensitivity signatures at the transcript level based on significant p values. However, the potential of sensitivity biomarkers has not been sufficiently understood because these investigations have focused on individual biomarkers and have not been carried out at the systems level. In this study, we applied a meta-analytical framework to compute co-expression between isoform pairs in two large datasets of RNA-seq profiles of breast cancer cell lines. We then built hallmark-related direct (HRD) networks by integrating a breast cancer specific isoform co-expression (BCIC) network and hallmark-related isoforms. Next, we explored the associations between isoform biomarkers and the functional clusters of the HRD network. The crucial isoform-based biomarkers for drugs were identified by functional clusters analysis and elucidated by combining isoform expression profiles with clinical information for breast cancer in The Cancer Genome Atlas.

Introduction Molecular MRD assays targeting NPM1 mutant (mut) transcripts have an established role for monitoring treatment efficacy in patients with NPM1mut AML. Approximately 25% of NPM1mut patients show persistent MRD level in the bone marrow (BM) at the end of treatment (EOT), which is associated with a higher risk of relapse (Ivey, NEJM 2016; Kronke, JCO 2011). Molecular persistence at low copy number (MP-LCN) is defined by the European LeukemiaNet (ELN) as MRD positivity in patients in morphological complete remission (CR) with Methods Consecutive patients with newly diagnosed NPM1mut AML receiving at least 2 cycles of intensive chemotherapy were included if MRD remained detectable at EOT. UK patients were all enrolled in the NCRI AML17 and AML19 studies. Australian patients were retrospectively identified. BM samples were analyzed by either RT-qPCR (n=75) (Ivey, NEJM 2016) or ultra-deep next-generation sequencing (UD-NGS, n=5, excluded from Cox model). We applied the ELN definitions for molecular progression or relapse (considered together as molecular failure [MF]), MP-LCN and complete molecular remission (CRMRD-). Kaplan Meier survival estimates were calculated from EOT to death (OS), and/or morphologic relapse (RFS), and/or MF (EFS). The optimal NPM1mut expression threshold was pre-determined by recursive partitioning. Variables with p Results A total of 80 patients had NPM1mut detectable at a median of 37 days (range 19-90; 12 [15%] were >60 days) from last course of chemotherapy; characteristics summarized in Table 1. The majority of patients (89%) had received 3-4 cycles of chemotherapy. At EOT, the median NPM1mut transcript level by RT-qPCR was 11 copies (range 0.3-8699; 3 were >2000) and median reduction from baseline was 4.7 log (range 2.1-6.0). A total of 34 patients (45%; 4 not monitored) fulfilled the criteria for MP-LCN. With a median follow-up time of 24 mo (range 8-77), 27 (34%) spontaneously achieved a sustained CRMRD- for >6 months, 31 (39%) had MF and 13 (16%) had morphologic relapse without preceding MF (due to rapid relapse [n=8], lack of monitoring [n=3], extramedullary or NPM1 wild-type relapse [n=1 each]) (Figure 1). Nine patients (11%) had persisting MP-LCN (included transient MRD negativity): median interval between EOT to the last known positive sample was 12 mo (range 3-32), and all patients currently remain alive and without morphologic relapse by the data cut-off date July 13, 2020. Next, we examined the variables associated with MF and/or morphologic relapse. In univariate analysis, factors associated (p Conclusions We describe the natural history of patients with NPM1 mutated AML who remain MRD positive after completing intensive chemotherapy. Almost half either spontaneously achieve MRD-negativity or have stable low-level expression without relapse with minimum follow up of 8 mo. Patients with both FLT3-ITD and Disclosures Tiong: Pfizer: Consultancy; Servier: Consultancy; Amgen: Consultancy, Honoraria. Dillon:Abbvie: Honoraria, Research Funding; Novartis: Honoraria; Menarini: Honoraria; Astellas: Honoraria; Pfizer: Honoraria, Research Funding; Jazz Pharmaceuticals: Honoraria; Amgen: Honoraria, Research Funding. Bajel:Abbvie: Honoraria; Pfizer: Honoraria; Amgen: Honoraria, Speakers Bureau; Novartis: Honoraria; Astellas: Honoraria. Russell:Pfizer: Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Speakers Bureau; Astellas: Membership on an entity's Board of Directors or advisory committees; Jazz: Research Funding, Speakers Bureau. Wei:MacroGenics: Consultancy, Honoraria; Pfizer: Honoraria; Servier: Consultancy, Honoraria, Research Funding; Walter and Eliza Hall Institute: Other: former employee and receives a fraction of its royalty stream related to venetoclax; Celgene: Consultancy, Honoraria, Research Funding; AbbVie Inc.: Consultancy, Honoraria, Research Funding; Genentech: Honoraria; Astra Zeneca: Honoraria, Research Funding; Janssen: Consultancy, Honoraria; Astellas: Consultancy, Honoraria; Novartis: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding.

Acute myeloid leukemia (AML) with t(8;21) is a heterogeneous disease. Although the detection of minimal residual disease (MRD), which is indicated by RUNX1‐RUNX1T1 transcript levels, plays a key role in directing treatment, risk stratification needs to be improved, and other markers need to be assessed. A total of 66 t(8;21) AML patients were tested for aldehyde dehydrogenase (ALDH) activity by flow cytometry at diagnosis, and 52 patients were followed up for a median of 20 (1‐34) months. The median percentage of CD34+ALDH+, CD34+CD38‐ALDH+, and CD34+CD38+ALDH+ cells among nucleated cells were 0.028%, 0.012%, and 0.0070%, respectively. The CD34+ALDH+‐H, CD34+CD38‐ALDH+‐H, and CD34+CD38+ALDH+‐H statuses (the percentage of cells that were higher than the individual cutoffs) were all significantly associated with a lower 2‐year relapse‐free survival (RFS) rate in both the whole cohort and adult patients (P = .015, .016, and .049; P = .014, .018, and .032). Patients with, ALDH status at diagnosis may improve MRD‐based risk stratification in t(8;21) AML, and concurrent high levels of CD34 + ALDH+ at diagnosis and MRD predict relapse.

Dysregulation of the alternative NF-κB signaling has severe developmental consequences that can ultimately lead to oncogenesis. Pivotal for the activation of the alternative NF-κB pathway is the stabilization of the NF-κB-inducing kinase (NIK). The aim of this review is to focus on the emerging role of NIK in cancer. The documented subversion of NIK in cancers highlights NIK as a possible therapeutic target. Recent studies show that the alterations of NIK or the components of its regulatory complex are manifold including regulation on the transcript level, copy number changes, mutations as well as protein modifications. High NIK activity is associated with different human malignancies and has adverse effects on tumor patient survival. We discuss here research focusing on deciphering the contribution of NIK towards cancer development and progression. We also report that it is possible to engineer inhibitors with high specificity for NIK and describe developments in this area.

Testosterone can induce impulsivity, a behavioral impairment associated with various psychiatric illnesses. The molecular mechanisms associated with testosterone-induced impulsivity are unclear. Our earlier studies showed that supraphysiological doses of testosterone to rats induced impulsive behavior, impacted hypothalamic-pituitary-adrenal axis (HPA) and hypothalamic-pituitary-gonadal axis interactions, and altered α2A adrenergic receptors in prefrontal cortex (PFC). Owing to the importance of GABAergic system in impulsivity and memory, the present study examines whether testosterone-mediated impulsivity is associated with changes in the expression of Gamma-Aminobutyric Acid (GABA) A and B receptor subunit transcripts (Gabra1, Gabra2, Gabra2 transcript variant 2, Gabra3, Gabra4, Gabra5, Gabra6, Gabrb1, Gabrb2, Gabrb3, Gabrg1, Gabrg2, Gabrg3, Gabbr1, Gabbr2) in rat PFC, and whether testosterone influences GABAA receptor subunit organization. We studied GABA receptor functions by examining GABA receptor-mediated calcium/calmodulin-dependent kinase signaling genes (Calm1, Calm2, Calm3, Camk2a, Camk2b, Camk2g, Camk2d, Camk4) in the testosterone-induced impulsivity model. Rats were left untreated as controls (C), gonadectomized (GDX), or GDX and injected with supraphysiological doses of testosterone (T). Impulsive behavior was examined using the go/no-go paradigm. Gene expression was studied using qRT-PCR and GABAA subunit reorganization using cross correlation. Our findings show that expressions of select GABAA receptor subunits (Gabra3, Gabra5, Gabra6) were significantly upregulated in PFC of T group compared to GDX or C groups. GABAA receptor subunit organization was different in C, T, and GDX groups. Additionally, Camk4 expression was significantly downregulated in T compared to C group. Our findings suggest that specific GABAA receptor subunit expression, their reorganization, and Camk4-mediated functions may be associated with testosterone-mediated impulsivity.

The root of Phlomis umbrosa has traditionally been used as a medicine in South Asian nations to treat colds and bone fractures, to staunch bleeding, and as an anti-inflammatory, and such use continues today. We identified 10 genes that are involved in terpenoid biosynthesis, while using the Illumina/Solexa HiSeq2000 platform. We investigated the transcript levels of the 10 genes using quantitative real-time PCR and quantified the level of terpenoid accumulation in different organs of P. umbrosa while using high-performance liquid chromatography. The transcript levels of PuHDR and PuHMGR1 were the highest among the studied genes. Sesamoside, an iridoid glycoside, appeared in higher quantity than shanzhiside methylester, umbroside (8-O-acetyl shanzhiside methyl ester), and acteoside. We speculate that PuHDR and PuHMGR1 may contribute to terpenoid biosynthesis in P. umbrosa. This study highlights the molecular mechanisms that underlie iridoid glycoside biosynthesis in P. umbrosa.

Core-binding factor acute myeloid leukemia is characterized by t(8;21) or inv(16) and the fusion proteins RUNX1-RUNX1T1 and CBFB-MYH11. International guidelines recommend monitoring for measurable residual disease every 3 months for 2 years after treatment. However, it is not known whether serial molecular monitoring can predict and prevent morphological relapse. We conducted a retrospective singlecenter study of 114 patients in complete remission who underwent molecular monitoring with real-time quantitative polymerase chain reaction analysis of RUNX1-RUNX1T1 or CBFB-MYH11 transcripts every 3 months. Morphological relapse was defined as re-emergence of >5% blasts and molecular relapse as ≥1 log increase in transcript level between two samples. Over a median follow-up time of 3.7 years (range, 0.2-14.3), remission persisted in 71 (62.3%) patients but 43 (37.7%) developed molecular or morphological relapse. Patients who achieved

Ras-related dexamethasone-induced 1 (RASD1) is abnormally expressed in many solid cancers. However, its potential role in adults with B-cell acute lymphoblastic leukemia (B-ALL) is unclear. Therefore, we aim to clarify the abnormal expression of the tumor-associated biomarker, RASD1, as a potential target for diagnosis and prognosis in adult Philadelphia-negative B-ALL. The expression of RASD1 was detected with RT-qPCR in 92 adults with de novo Ph-negative B-ALL and 40 healthy controls. The correlation between RASD1 transcript levels and relapse was assessed. RASD1 transcript levels in patients with Ph-negative B-ALL (median 81.76%, range 0.22%-1824.52%) were significantly higher than those in healthy controls (7.59%, 0.46%-38.66%; PRASD1 transcript levels had a lower 5-year relapse-free survival (RFS, 47.5% [32.9%, 62.1%] vs. 63.1% [49.0%, 77.2%]; P = 0.012) and a higher 5-year cumulative incidence of relapse (CIR, 52.0% [37.4%, 66.6%] vs. 36.2% [22.2%, 50.2%]; P = 0.013) especially in patients receiving chemotherapy only. Multivariate analysis showed that a low RASD1 transcript level was an independent risk factor for RFS (HR = 2.938 [1.427, 6.047], P = 0.003) and CIR (HR = 3.367 [1.668, 6.796], P = 0.001) in patients with Ph-negative B-ALL. RASD1 transcript levels were significantly higher in patients with Ph-negative B-ALL and a low RASD1 transcript level was independently correlated with increased relapse risk.

The majority of patients with TKI failure respond to HCT. However, the relapse risk remains high. This study has evaluated transplant outcomes in 223 CML patients with TKI failure due to resistance (n = 132) or intolerance (n = 29), as well as those that were TKI naïve/responding with advanced disease (n = 35) or with chronic phase (CP, n = 27). We studied outcomes according to post-transplant BCR-ABL transcript level within 3 months. With respect to transplant outcomes according to the post-transplant BCR/ABLtranscript level within 3 months, the group failing to achieve a 1.3 log reduction (n = 14, 12.4%) showed the highest relapse rate of 78.6% at 5 years, compared to 26.2% and 24.1% in the groups achieving 1.3-4.0 log reduction (n = 45, 39.8%), and ≥4.1 log reduction (n = 54, 47.8%) respectively (p 0.001). Multivariate analysis confirmed that the group failing to achieve a 1.3 log reduction had a 2.3-fold higher risk of death and 6.6 times higher risk of relapse. Poor overall survival after HCT was associated with advanced disease at diagnosis, but not disease status prior to HCT. Of 61 patients who relapsed after HCT, 47 were treated with post-transplant TKI therapy; those receiving TKI after loss of MR2 or MMR showed higher rates of response and survival compared to those receiving TKI after hematologic relapse (p 0.001). QPCR log reduction level within 3 months post transplantation is prognostic in this population.

Cassia tora is a plant of medicinal importance. Medicinal plants from different localities are believed to differ in their therapeutic potency. In this study, six populations of C. tora with different eco-geographical origins were investigated genotypically (ISSR) and phytochemically (FTIR) to establish an integrated approach for population discrimination and authentication of the origin of this medicinal herb. CHS gene expression analysis and determination of flavonoid content were carried out to substantiate the study. A total of 19 population-specific authentication bands were observed in 11 ISSR fingerprints. Authentication codes were generated using six highly polymorphic bands, including three authentication bands. FTIR spectra revealed that the peaks at wavenumber 1623 cm−1 (carbonyl group) and 1034 cm−1 (>CO- group) were powerful in separating the populations. These peaks are assigned to flavonoids and carbohydrates, respectively, were more intense for Ranchi (highland) population. Variation in the transcript level of CHS gene was observed. The findings of FTIR and RT-PCR analyses were in agreement with the TFC analysis, where, the lowest amount of flavonoids observed for Lucknow (lowland) population. All the populations of C. tora have been authenticated accurately by ISSR analyses and FTIR fingerprinting, and the Ranchi site was observed to be more suitable for the potential harvesting of therapeutic bioactive compounds.

Hyoscyamine (Hyos) and scopolamine (SCP) are drugs widely used as antimuscarinic to treat diseases such as Parkinson’s or to calm schizoid patients. In this study, with the aim of enhancing tropane...

Long non-coding RNAs affect cancer progression and gastric cancer(GC) tumorigenesis impressively. We investigated the expression of a novel long intergenic non-coding RNA—lincRNA-AC064834.1(ENST00000413290.1)—in GC patients. LincRNA-AC064834.1 was selected from the differentially expressed lncRNA transcripts by RNA-seq results. Quantitative RT-PCR examined the lincRNA-AC064834.1 expression in GC tissues and adjacent noncancerous tissues(ANCTs) (n = 80 samples). The association between the expression level of AC064834.1 and clinicopathology was tested. In addition, the suitability of the transcript level of this gene in differentiation of tumoral from non-tumoral tissues was assessed by plotting the ROC curve. The results showed that the AC064834.1 relative expression levels were considerably higher in tumor tissues than in ANCTs (Fold change/log2 standard error, 4.52 (0.22–73.77); p = 0.006). The overall sensitivity and specificity of AC064834.1 in GC diagnosis were 67.5%. RNA expression level of AC064834.1 did not significantly correlate with clinicopathologic characteristics. The lincRNA-AC064834.1 might serve as a novel potential biomarker for GC.

Background: The Canadian tyrosine kinase inhibitor (TKI) discontinuation trial, named "Treatment Free Remission Accomplished By Dasatinib " (NCT#02268370), has reported 56.8% molecular relapse-free survival (mRFS) rate at 12 months after imatinib (IM) discontinuation. MMR loss occurred quickly after IM discontinuation, typically within 2-4 months, while those who lost MR4 on two consecutive measurements tended to lose their molecular response more gradually. BCR-ABL transcript doubling time (DT) after TKI discontinuation is a reciprocal concept to transcript halving time following TKI therapy. Due to inter-individual differences in DT after TKI discontinuation, DT can be used as a potential biomarker to identify those patients at high-risk for TFR failure when measured before they experience a clinically significant event of molecular relapse. The present study has not only evaluated the kinetics of BCR-ABL transcript rise after IM discontinuation, but also explored the predictive/prognostic role of DT of BCR-ABL transcript level as an early predictor of TFR failure. Patients and methods: Changes of BCR-ABL1 transcript level in each patient were assessed monthly by estimating the number of days required for BCR-ABL1 to double from the previous expression level/measurement, termed the DT. Based on the BCR-ABL1 qPCR value taken monthly in the first 6 months after IM discontinuation, DT was calculated monthly, as x = ln(2)/K, where x is the DT and K is the fold BCR-ABL1 change from the previous value divided by the number of days between each measurement. K was determined as follows: K = [ln(b) - ln(a)]/t, where a is the BCR-ABL1 value of the previous measurement, b is the BCR-ABL1 at the relevant time point, and t is the number of days between measurements. The baseline qPCR level from the prior month to TKI discontinuation was referenced. The distribution of DT was assessed at each time point of DT measurement within the first 6 months. In order to define cut-off levels for BCR-ABL1 qPCR and DT for the first 6 months, multiple statistical parameters were taken into account including positive (PPV) and negative predictive value (NPV), accuracy and F1 score of DT value, resulting in the DT value of 12.75 days at 2 months as the optimal cut-off value of DT value. Patients were stratified into the 3 groups based on the DT value of 12.75 days at 2 months after IM discontinuation. The high-risk group was defined as the patients showing DT < 12.75 days but above 0, i.e. rapidly proliferating CML cells, implying a high risk for TFR failure with a shorter DT. The intermediate-risk group was defined as those patients with DT ≥ 12.75 days, i.e. more slowly proliferating CML cells implying intermediate risk for TFR failure. The low-risk group was defined as patients showing DT of zero or below, i.e. no increase in the size of the pool of cells expressing BCR-ABL, implying a low risk for TFR failure. The mRFS was analyzed for each of these risk groups at 6 monthly intervals after TKI discontinuation. Results: We compared the DT values of the patients that failed TFR with those from the patients who maintained their molecular response at last follow-up. The DT values at 2 months were much shorter in patients who failed TFR after IM cessation (median 8.32 days) compared to those who maintained molecular response (median 20.7 days; p The DT value of 12.75 days was defined as the optimal value for DT at 2 months with the NPV, PPV, accuracy and F1 score of 80.90%, 96.43%, 84.62% and 0.75, respectively, as the -log10(p-value), accuracy and F1 score reached a plateau at a DT of 12.75 days as presented in the Figure A. At a DT value of 12.75 days at 2 months after IM discontinuation, patients were stratified into 3 groups: high- (n=26), intermediate- (n=16) and low-risk groups (n=71; Figure B). With respect to mRFS rate, the high-risk group showed 7.7% mRFS rate at 12 months compared to 53.6% in the intermediate-risk group or 90.0% in the low-risk group (p Conclusion: Monthly assessment of DT based on the monthly BCR-ABL qPCR is useful to identify the patients with an imminent risk of molecular recurrence after IM cessation for TFR. Figure Disclosures Bence-Bruckler: Merck: Membership on an entity's Board of Directors or advisory committees. Keating:Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees; Hoffman La Roche: Membership on an entity's Board of Directors or advisory committees; Sanofi: Membership on an entity's Board of Directors or advisory committees; Seattle Genetics: Consultancy; Merck: Membership on an entity's Board of Directors or advisory committees; Janssen: Membership on an entity's Board of Directors or advisory committees; Servier: Membership on an entity's Board of Directors or advisory committees; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Shire: Membership on an entity's Board of Directors or advisory committees; Taiho: 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. Busque:Novartis: Honoraria; BMS: Honoraria; Pfizer: Honoraria. Delage:Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding. Lipton:BMS: Consultancy, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; Bristol-Myers Squibb: Honoraria; Ariad: Consultancy, Research Funding; Pfizer: Consultancy, Honoraria, Research Funding; Novartis: Consultancy, Research Funding. Leber:Pfizer: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Abbvie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; BMS/Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees; Treadwell: Honoraria, Membership on an entity's Board of Directors or advisory committees; Takeda/Palladin: Honoraria, Membership on an entity's Board of Directors or advisory committees; Alexion: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Lundbeck: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Otsuka Pharmaceutical: Honoraria, Membership on an entity's Board of Directors or advisory committees.

The long term effects of fish oil (FO) substitution by increasing the levels of vegetable oils (VO), 0% (CTR), 50% (VO50) and 100% (VO100), in diets for Senegalese sole were evaluated in terms of skeletal muscle cellularity and expression of related genes. After 140 days of feeding, all fish had similar body weight and length. The inclusion of 50% VO did not result in differences in muscle cellularity, but dorsal muscle cross-sectional area and fast-twitch fibre diameter increased in fish fed total FO substitution, whilst fibre density was reduced (P < 0.05) in relation to CTR. The total number of fibres was similar in all treatments. FO substitution did not affect the transcript levels of myogenic genes (myf5, mrf4, myog, myod1, myod2), but resulted in a two-fold increase of fgf6 transcript levels compared to CTR (P < 0.05). The relative expression of igf-I was higher in VO100 than in VO50, but was similar to CTR. FO substitution resulted in cellularity changes related to the stimulation of muscle hypertrophic growth, but not hyperplastic growth, and associated with a nutritional modulation of fgf6 by dietary VO. This study indicates that 50% VO does not affect the muscle phenotype, but total FO substitution stimulates muscle hypertrophy. © 2017 The Royal Society of Chemistry. This work was partially supported by NORTE-07-0124-FEDER-000038, in the context of the North Region Operational Programme (ON.2-O Novo Norte), under the project Sustainable Aquaculture and Animal Welfare (AQUAIMPROV) and the FCT research strategic funding UID/Multi/04423/2013.

Precisely determining the postmortem interval (PMI) is crucial to civil, criminal and forensic cases. A technique to exploit the postmortem RNA transcript level was developed to increase the accuracy and practicality of PMI estimation. For this purpose, lung tissues and muscle tissues were removed at twelve time points (0–144 h) from rat corpses that had been stored at three different temperatures (10, 20 and 30 °C). Human tissues were collected at autopsy from twelve real cases with known PMI values and other parameters. After the RNA was extracted from all these samples, the transcript levels of nine biomarkers were analyzed by real-time quantitative PCR (RT-qPCR). With the assistance of geNorm, miR-195 , miR-200c , 5S , U6 and RPS29 were selected as reference biomarkers for lung specimens; miR-1 , miR-206 , 5S and RPS29 were chosen as control markers for muscle tissues. On the contrary, ACTB and GAPDH were significantly correlated with the PMI. The mathematical models using these target biomarkers were constructed to describe the characteristic relationship between △Ct values (normalized to reference biomarkers) and the observed PMI for each temperature group. Following validation, the relatively low error rates (7.4% and 12.5% for rat and human samples, respectively) demonstrated the accuracy and reliability of the mathematical model. We believe these results indicate that the multi-parametric mathematical model can become a practical tool for PMI estimation.

The aim of this study was to determine the influence of two types of UV-A LEDs on the growth and accumulation of phytochemicals in kale (Brassica oleracea var. acephala). Fourteen-day-old kale seedlings were transferred to a growth chamber and cultivated for 3 weeks. The kale plants were subsequently subjected to two types of UV-A LEDs (370 and 385 nm) of 30 W/m2 for 5 days. Growth characteristics were all significantly increased in plants exposed to UV-A LEDs, especially at the 385 nm level, for which dry weight of shoots and roots were significantly increased by 2.22 and 2.5 times, respectively, at 5 days of treatment. Maximum quantum efficiency of photosystem II photochemistry (Fv/Fm ratio) began to decrease after 3 h of treatment compared to the control. The total phenolic content of plants exposed to the two types of UV-A LEDs increased by 25% at 370 nm and 42% at 385 nm at 5 days of treatment, and antioxidant capacity also increased. The two types of UV-A LEDs also induced increasing contents of caffeic acid, ferulic acid, and kaempferol. The reactive oxygen species (ROS) temporarily increased in plants exposed to the two types of UV-A LEDs after 3 h of treatment. Moreover, transcript levels of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), and flavanone 3-hydroxylase (F3H) genes and PAL enzyme activity were higher in plants treated with UV-A LEDs. Our results suggested that short-term UV-A LEDs were effective in increasing growth and improving antioxidant phenolic compounds in kale, thereby representing a potentially effective strategy for enhancing the production of phytochemicals.

Background: Baricitinib (bari) is an oral selective Janus kinase (JAK)1 and JAK2 inhibitor. In the Phase 2 study, JAHH (NCT02708095), treatment with bari resulted in significant improvements in patients (pts) with active systemic lupus erythematosus (SLE) receiving standard background therapy, compared with placebo (PBO).1 Objectives: To use global gene expression to characterize baseline (BL) gene expression among SLE pts in JAHH and to describe the mechanism of action (MoA) of bari during treatment in active SLE. Methods: A total of 314 pts were randomized 1:1:1 to PBO, bari 2-mg, or bari 4-mg once daily for 24 weeks (Wks) in JAHH. RNA isolated from whole blood at BL and Wks 2, 4, 12, and 24 was analyzed using HTA2.0 arrays, possessing over 925,000 specific individual probes. Data were summarized to transcript level and analyzed using a mixed effects model on a log2 transformed response with multiplicity correction. Results: Baseline gene expression analysis comparing healthy controls to SLE pts revealed elevated expression of genes involved in innate and adaptive immune pathways and biologic processes thought to be key to SLE pathogenesis. Baseline findings from the ILLUMINATE trial of tabalumab (1760 SLE pts at BL)2 were independently replicated, including individual gene-specific changes in the SLE populations versus controls. Treatment-induced changes were predominantly observed in the bari 4-mg versus PBO group at early time points (Wks 2 and 4; bari 4-mg > 2-mg). Statistically significant changes were observed for type I and II IFN responsive genes, and in the JAK/signal transducer and activator of transcription (STAT) canonical and noncanonical signaling pathways, in the bari 4-mg versus PBO group comparisons. There were biologically notable and statistically significant bari-associated decreases in gene expression from BL, compared to PBO, for interleukin (IL)-3, -5, -6, -7, and granulocyte macrophage colony-stimulating factor pathways, attributable to inhibition of JAK/STAT signaling. Conclusion: These results build upon previous studies of gene expression in ILLUMINATE, comparing SLE at BL with healthy controls; they advance our understanding of lupus pathogenesis, and further delineate the MoA of bari in SLE. These data independently confirm the marked elevations of specific type I and II IFN genes at BL in clinical trial pts on standard of care treatment. Changes in gene expression with bari impacted IFN responsive genes as well as JAK/STAT signaling pathways. Interpretation of results is limited by sample size and statistical power, and presence of concomitant medications. Importantly, gene expression may not translate directly to protein expression; however, ongoing proteomic studies (including specific assays for type I IFN, IFN-γ, IL-12, IL-17, and IL-23) will help extend these gene expression results and further describe the MoA of bari in SLE. References: [1] Wallace D, et al. Lancet. 2018;392:222-231 [2] Hoffman R, et al. Arthritis Rheumatol. 2017;69:643-654 Disclosure of Interests: Thomas Dorner Grant/research support from: Eli Lilly, Janssen, Roche, UCB Pharma, Consultant for: Eli Lilly, Janssen, Roche, UCB Pharma, Speakers bureau: Eli Lilly, Janssen, Yoshiya Tanaka Grant/research support from: Abbvie, Astellas, Bristol-Myers Squibb, Chugai, Daiichi-Sankyo, Eisai, Mitsubishi-Tanabe, MSD, Ono, Taisho-Toyama, Takeda, Speakers bureau: Abbvie, Asahi-kasei, Astellas, Bristol-Myers Squibb, Chugai, Daiichi-Sankyo, Eli Lilly, Eisai, Glaxo-Smithkline, Janssen, Mitsubishi-Tanabe, Novartis, Pfizer Japan Inc, Sanofi, Takeda, UCB, YL Biologics, Michelle A Petri Shareholder of: Pfizer, Merck, Grant/research support from: AstraZeneca, Exagen, Consultant for: Eli Lilly, GSK, Merck EMD Serono, Janssen, Amgen, Novartis, Quintiles, Exagen, Inova Diagnostics, AstraZeneca, Blackrock, Glenmark, UCB, and the Annenberg Center for Health Sciences, Josef S. Smolen Grant/research support from: AbbVie, Eli Lilly, Janssen, MSD, Pfizer Inc, Roche, Consultant for: AbbVie, Amgen, AstraZeneca, Astro, Celgene, Celtrion, Eli Lilly, GlaxoSmithKline, ILTOO, Janssen, Medimmune, MSD, Novartis-Sandoz, Pfizer Inc, Roche, Samsung, Sanofi, UCB, Speakers bureau: AbbVie, Amgen, AstraZeneca, Astro, Celgene, Celtrion, Eli Lilly, GlaxoSmithKline, ILTOO, Janssen, Medimmune, MSD, Novartis-Sandoz, Pfizer Inc, Roche, Samsung, Sanofi, UCB, Ernst Dow Shareholder of: Eli Lilly and Company, Employee of: Eli Lilly and Company, Richard E Higgs Shareholder of: Eli Lilly and Company, Employee of: Eli Lilly and Company, Guilherme Rocha Shareholder of: Eli Lilly and Company, Employee of: Eli Lilly and Company, Robert Benschop Shareholder of: Eli Lilly and Company, Employee of: Eli Lilly and Company, Maria Silk Shareholder of: Eli Lilly and Company, Employee of: Eli Lilly and Company, Stephanie de Bono Shareholder of: Eli Lilly and Company, Employee of: Eli Lilly and Company, Robert Hoffman Shareholder of: Eli Lilly and Company, Employee of: Eli Lilly and Company

Gallbladder carcinoma (GBC) is a malignant tumor in gastrointestinal system. In this review, we mainly focus on three molecular levels to illuminate the potential molecular mechanisms in gallbladder carcinoma. First, we review genes with mutation and methylation associated with occurrence and development of GBC. Second, we review non-coding RNA and key differential genes at the transcript level, especially for their interactions. Third, we review crucial proteins in GBC. Moreover, we also discuss the challenges of these molecular signatures in clinical applications. Finally, we discuss potential application of these crucial genes in prevention, diagnosis and treatment of GBC.

Background. The current approach to treatment of acute myeloblastic leukemia (AML) includes the achievement of maximum tumor reduction and, therefore, eradication of a leukemic clone. The goal of the therapy is to achieve undetectable levels of the target gene, except an isolated molecular rearrangement of RUNX1-RUNX1T1. Aim. To estimate the dynamics of the RUNX1-RUNX1T1 level and relevant clinical manifestations during the monitoring of various stages of the program therapy and after its completion. Methods. The article presents a description of 10 cases of AML with isolated RUNX1-RUNX1T1 expression (n = 4) and the expression in combination with different molecular and cytogenetic abnormalities (n = 6). In addition, a long-term monitoring of the gene expression by quantitative determination of RUNX1-RUNX1T1 using a real-time PCR was presented. Results. The incidence of relapses in a group with a decreased RUNX1-RUNX1T1 expression level of >2 log is 75 % as compared to patients with a less significant reduction of the transcript level (with the relapse incidence equal to 0 %) (p = 0.05). The increase of the RUNX1-RUNX1T1 level against the background of bone marrow remission by more than 1 log coincided with a bone marrow relapse within 5-18 weeks. In addition, long-term persistence of a certain transcript level after the completion of a program therapy without relapse is possible. Conclusion. The study analyzed possible molecular background of different clinical outcomes of long-term persistence of the RUNX1-RUNX1T1 transcript that might lead to an individualized approach to AML patients.

The present study was performed to determine whether the adherence to regular follow-up assessments using standardized real-time quantitative polymerase chain reaction (qPCR) and/or cytogenetic tests in Lebanese patients with chronic myeloid leukemia (CML) meet the European LeukemiaNet recommendations. The present study was a retrospective analysis of 34 patients diagnosed with chronic phase CML who had been treated with tyrosine kinase inhibitors and monitored with regular cytogenetic tests and/or measurement of the BCR-ABL transcript level at 3, 6, and 12 months from 2006 until 2015 in 3 university hospitals in Lebanon. All patients were included and monitored in an adherence program (SAWA program). The male/female ratio was 3:1. The median age was 50 years, and the mean age was 50 years. As frontline treatment, 29 patients started imatinib and 5 patients received second-generation tyrosine kinase inhibitors. We defined compliance to the monitoring tests as regulary realizing the qPCR at 3, 6, and 12 months. Of the 36 patients, 15 underwent the recommended tests at 3, 6, and 12 months, representing a compliance rate of 41.6%; 28 of the 34 patients underwent the recommended tests only twice in the first follow-up year. Only 14 patients underwent qPCR at 3 months. We believe that despite the inclusion of our patients in an adherence program, the compliance rate is still low. We also believe that greater effort is required to improve the adherence to regular follow-up examinations.

Homoharringtonine combined aclarubicin and cytarabine (HAA) has been demonstrated to achieve a high remission rate and provide a survival advantage in acute myeloid leukemia (AML). To investigate whether HAA is an ideal induction regimen for t(8;21)AML, we retrospectively analyzed the data of 140 patients from the last 8 years in our center. When achieving complete remission (CR), the post-remission treatment was administered as a minimal residual disease-directed risk-stratification treatment protocol. The RUNX1/RUNX1T1 transcript level was assessed by RT-qPCR. The last follow-up was conducted in October 2015. In total, thirty patients received an HAA regimen as the induction treatment. The CR rate after one cycle of the HAA regimen was 93.3% (28/30). One patient achieved partial remission, and one had no response. No patients died during induction treatment. The median fold decrease of the RUNX1/RUNX1T1 transcript level was 200 (1-358000), and 16.7% (5/30) patients achieved >3 log decrease after one cycle of the HAA regimen. The estimated 4-year disease-free survival and overall survival were 89.9% and 90.8%, respectively. We concluded that the HAA regimen is highly effective as the first course of induction therapy for t(8;21) AML, and this needs to be confirmed in a large population in the future.

The aim of this study was to clarify the clarithromycin resistance mechanisms of β-lactamase-nonproducing ampicillin-resistant Haemophilus influenzae strains. In all clarithromycin-resistant strains, the transcript level of acrB was significantly elevated, and these strains had a frameshift mutation in acrR . Introduction of the acrR mutation into H. influenzae Rd generated a clarithromycin-resistant transformant with the same MIC as the donor strain. Our results indicate that the acrR mutation confers clarithromycin resistance by the increasing the transcription of acrB .

The influence of cadmium sulphate (100 µM) on GS3 gene expression encoding glutathionesynthetase – one of the main enzymes of glutathione synthesis, in wheat seedlings roots and leaves was investigated. It was showed that after 1 hour of cadmium effect on wheat seedlings its accumulation was observe with increase level of GS3 transcripts. In leaves the mRNA content rapidly increased (after 30 min) after beginning of experiment but the cadmium accumulation was after 1 day. The higher transcripts level of GS3 in leaves and roots saved for 7 days of experiment. The analysis of MDA content in roots showed that its accumulation in roots didn't observed but in leaves the MDA content increased under long cadmium treatment (3–7 days) on seedlings. It may suggest about activation and effective antioxidant defense system, including glutathionesynthetase in cadmium presence. Also the increased level electrolyte leakage in leaves wasn't observed, in consequence the membrane permeability didn’t damage. Taking all results together we can conclude that cadmium in concentration 100 µM didn't lead to damage on wheat seedlings and accumulation of transcript level of GS3, encoding glutathionesynthetase is the part of adaptation, what support to survive and save to life-sustaining activity of wheat under cadmium presence in environment.

The basic helix-loop-helix (bHLH) family proteins, a group of functionally diverse transcription factors (TFs), control a series of plant biological processes. However, the involvement of bHLH TFs in fruit ripening still rarely reported. In this study, a banana fruit bHLH TF, named as MabHLH7, was identified and characterized. MabHLH7 was ethylene-inducible and nuclear-localized, and its transcript level was increased during banana fruit ripening. More importantly, MabHLH7 can directly bind to and activate the promoters of several cell wall-modifying-related genes, including MaXTH12, MaEXP2/21, MaPME4/5, MaPG4, and MaPL1/2, which were significantly enhanced in the ripening stage. Overall, our findings establish a transcriptional regulatory network during banana fruit ripening, in which MabHLH7 activates a small subset of cell wall-modifying-related genes through directly binding to their promoters.

Indoleamine 2,3-dioxygenase (IDO) is a negative regulator of the immune system. To approach reasons of variability in the generation of anti-HBs antibodies in response to HBV vaccination among hemodialysis (HD) subjects, we aimed to investigate whether the IDO gene (IDO1) transcript levels are associated with post-vaccination anti-HBs production and IDO1 polymorphic variants.The IDO1 transcript was determined by qRT-PCR analysis in 110 HD patients. IDO1 (rs3739319, rs9657182) genotyping was carried out by HRM analysis.The relative IDO1 transcript levels were not associated with IDO1 polymorphic variants. There were 16 non-responders (not able to produce anti-HBs10 IU/L), 74 patients with anti-HBs 10-999 IU/L, and 20 hyperactive responders (anti-HBs ≥1000 IU/L). IDO1 transcript levels were different among these groups (0.832, 0.423-4.373; 1.114, 0.317-6.582; 0.680, 0.164-3.014; respectively, Kruskal-Wallis P = 0.024). Significance in IDO1 transcript was shown between anti-HBs titers 10-999 IU/L and ≥1000 IU/L (P = 0.020). IDO1 transcript level0.743 indicated 3.38 (1.17-9.72) higher probability of hyperactive immunization (adjusted P = 0.005).In HD patients, ability to generate anti-HBs is not associated with IDO1 transcript levels. Hyperactive anti-HBs responses occur in patients showing lower IDO1 transcript. The latter cannot be predictable by genotyping IDO1 rs3739319 or rs9657182.

For patients with chronic myeloid leukaemia (CML), treatment guidelines recommend monitoring response to treatment with tyrosine kinase inhibitors (TKIs) by testing the BCR-ABL1 fusion gene transcript level using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). Despite recent efforts to standardise protocols for BCR-ABL1 testing, some variability remains among laboratories in the UK regarding the techniques used and the approach to reporting results. This increases the risk of misinterpretation of results by both clinicians and patients. An expert panel met to discuss current issues surrounding BCR-ABL1 testing in the UK and to develop guidance for laboratories, with emphasis on the optimal approach to reporting laboratory results. Topics included the minimum required information to include in the laboratory report, units of measurement, test sensitivity, and BCR-ABL1 transcript variants. To aid communication between laboratories and clinics, standard forms were generated that could be used by 1) clinics when submitting samples to laboratories, and 2) laboratories when reporting results to clinics. Standardising the way in which BCR-ABL1 test results are reported from laboratories to clinics should help to improve communication, interpretation of results, and patient care.

The pathogenesis of diabetes mellitus is a complicated process involving much gene regulation. The molecular mechanism of mulberry (Morus alba L.) leaf in the treatment of diabetes is not fully understood. In this study, we used the Illumina HiSeq™ 2,500 platform to explore the liver transcriptome of normal mice, STZ-induced diabetic mice, and mulberry leaf-treated diabetic mice, and we obtained 52,542,956, 52,626,414, and 52,780,196 clean reads, respectively. We identified differentially expressed genes (DEGs) during the pathogenesis of diabetes in mice. The functional properties of DEGs were characterized by comparison with the GO and KEGG databases, and the results show that DEGs are mainly involved in the metabolic pathway. qRT-PCR was used to analyse 27 differential genes involved in liver expression in different groups of diabetic mice. Among the DEGs, the expression of Scube1, Spns3, Ly6a, Igf2, and other genes between the control (C) and diabetic control (DC) groups was significantly upregulated; the expression of Grb10, Mup2, and Fasn was significantly downregulated; the expression of the Sqle, Lss, and Irs2 genes between the C group and diabetic group treated with mulberry (DD) was significantly upregulated; the expression of Fabp2, Ly6a, and Grb10 was significantly downregulated; and the expression of Sqle and Lss was significantly upregulated in the DC and DD groups, but Tap1, Igf2, and Spns3 were significantly downregulated. The results of Western blot validation showed that dynamic changes in proteins, such as IGF2, Ly6a, Grb10, and UBD, occurred to regulate the incidence of diabetes by influencing the insulin receptor substrate (IRS) signaling pathway.

BACKGROUND: In chronic phase (CP) chronic myeloid leukemia (CML) nilotinib (NIL) showed better efficacy compared to imatinib. The higher rates of deep molecular response with NIL may translate into more patients (pts) eligible for treatment discontinuation. On the other hand, cardiovascular toxicity may limit NIL use in selected groups of pts (e.g. elderly pts). Long follow-up is required for a proper evaluation of the benefit-risk ratio between efficacy (including treatment-free remission [TFR] rates) and toxicity. However, only few data are currently available on the very long-term outcome of NIL treated pts. AIM: To describe the very-long term outcome of patients treated with nilotinib first-line. METHODS: The GIMEMA CML WP initiated in 2007 a phase II study (NCT 00481052) with NIL first-line (400 mg BID). Seventy-three adult (≥18 years old) pts with newly diagnosed, CP-CML were enrolled at 18 GIMEMA Centers in Italy. Median age was 51 (18-83) years. ELTS risk score was low in 64.4% of pts, intermediate in 30.1% of pts, and high in 5.5% of pts. Early and mid-term results have been previously published. Here, we report the very long-term outcome of this study, focusing on survival measures, TFR rates and adverse events. The updated, final results (with 10-year follow-up) of the trial will be available for presentation at the meeting. RESULTS: After a median follow-up of 92 months, 70 (95.9%) pts are alive and progression-free. Three pts (4.1%) died (progression to blast phase, n=1; CML-unrelated death, n=2). Failures according to ELN 2013 recommendation occurred in 5 (6.8%) pts (progression to BP, n=1; BCR-ABL transcript level > 1% at 12 months, n=2; confirmed loss of major molecular response, n=2). At last contact, 45 (61.6%) pts were still on NIL (dose ≥ 600 mg/day in 29 pts; 400 or 450 mg/day in 12 pts; ≤ 300 mg in 4 pts). Reasons for permanent NIL discontinuation in the remaining 28 (38.4%) pts were: adverse events (n=17, 23.3%; two of these pts maintained a deep molecular response without treatment - TFR); successful TFR attempts (n=9, 12.3%); failures (n=2 pts, 2.7%). The median age at CML diagnosis of pts obtaining the TFR was 47 (21-70) years. Fifteen athero-thrombotic events (ATEs) occurred in 13 (17.8%) pts (peripheral arterial occlusive disease, n=5; acute myocardial infarction, n=5; others ATEs, n=5), after a total NIL exposure of 478 years (3.1 events/100 pt-years). The median age at CML diagnosis of these pts was 66 (43 - 83) years; 8/13 (61.5%) pts had at least a baseline cardiovascular risk factor (hypertension, diabetes, hypercholesterolemia, or BMI ≥ 30). SUMMARY: These data highlight the efficacy of first-line NIL in the long-term, with excellent overall survival. TFR was obtained in 15% of pts, and particularly in younger ones. Some safety concerns remain, with ATEs occurring in 17.8% of pts (mainly in elderly ones), although none of these events was fatal. Disclosures Gugliotta: Incyte: Honoraria; Pfizer: Honoraria; Novartis: Honoraria. Castagnetti:Incyte: Honoraria; Pfizer: Honoraria; Bristol Myers Squiib: Consultancy, Honoraria; Novartis: Honoraria. Breccia:Novartis: Honoraria; Incyte: Honoraria; BMS: Honoraria; Pfizer: Honoraria; Celgene: Honoraria. Levato:BMS: Honoraria; Incyte: Honoraria; Pfizer: Honoraria; Novartis: Honoraria. Stagno:Pfizer: Honoraria; Novartis: Honoraria; BMS: Honoraria; Incyte: Honoraria. Tiribelli:Incyte: 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; Pfizer: Membership on an entity's Board of Directors or advisory committees. Bocchia:Incyte: Honoraria; BMS: Honoraria; Novartis: Honoraria. Cavazzini:Novartis: Honoraria; Incyte: Honoraria; Pfize: Honoraria. Caocci:Novartis: Honoraria; Celgene: Honoraria. Albano:Incyte: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees. Martinelli:Ariad: Consultancy, Other: trial grant; Amgen: Consultancy, Other: trial grant; Novartis: Consultancy, Other: trial grant; Pfizer: Consultancy, Other: trial grant; Janssen: Consultancy, Other: trial grant; Abbvie: Consultancy, Honoraria, Other: trial grant; Incyte: Consultancy, Other: trial grant; Daiichi Sankyo: Consultancy, Honoraria; Celgene: Consultancy, Honoraria, Other: trial grant; Roche: Consultancy, Other: trial grant. Soverini:Incyte: Consultancy. Foà:Abbvie: Consultancy, Speakers Bureau; Celltrion: Membership on an entity's Board of Directors or advisory committees; Amgen Inc.: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Incyte: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Janssen: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Roche: Consultancy, Speakers Bureau; Shire: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Abbvie: Consultancy, Speakers Bureau; Pfizer: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Pfizer: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Incyte: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Amgen Inc.: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Janssen: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celltrion: Membership on an entity's Board of Directors or advisory committees; Roche: Consultancy, Speakers Bureau; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Shire: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau. Pane:GSK: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees, Other: research founding; BMS: Membership on an entity's Board of Directors or advisory committees; Amgen: Membership on an entity's Board of Directors or advisory committees; Janssen: Membership on an entity's Board of Directors or advisory committees. Cavo:amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel accommodations, Speakers Bureau; janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: travel accommodations, Speakers Bureau; bms: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; sanofi: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; novartis: Honoraria; takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees. Saglio:Celgene: Consultancy; BMS: Consultancy; Novartis: Consultancy; Ariad: Consultancy; Incyte: Consultancy; Pfizer: Consultancy; Jansen: Consultancy. Baccarani:Novartis: Consultancy, Speakers Bureau; Incyte: Consultancy, Speakers Bureau; Takeda: Consultancy. Rosti:BMS: Speakers Bureau; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Pfizer: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Incyte: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.

Objectives Tigecycline non-susceptible Acinetobacter nosocomialis (TNAN) has been discovered in clinical isolates. The resistance-nodulation-cell division (RND)-type efflux system plays a major role in tigecycline non-susceptible Acinetobacter baumannii, but the mechanism in A. nosocomialis remains unknown. Our aim was to analyse the contribution of efflux-based tigecycline resistance in clinical A. nosocomialis isolates collected from multiple medical centres in Taiwan. Methods A total of 57 A. nosocomialis isolates, including 46 TNAN and 11 tigecycline-susceptible A. nosocomialis (TSAN) isolates, were analysed. Of these, 46 TNAN isolates were clustered to ST410 (43 isolates) and ST68 (three isolates) by multi-locus sequence typing. Results The relationship between the RND efflux pump and tigecycline resistance was indirectly verified by successfully reducing tigecycline resistance with NMP, an efflux pump inhibitor. The three RND efflux systems (AdeABC, AdeIJK and AdeFGH) were detected in all clinical isolates. The transcript level of adeB gene increased significantly and was correlated with tigecycline resistance. Moreover, the AdeRS two-component system was further classified into four different types of AdeRS patterns considering the amino acid sequence. Further analysis showed that tigecycline resistance was related to the transcript level of adeB gene and the AdeRS pattern. Conclusion This study showed that the dissemination of TNAN isolates in Taiwan is attributable mainly to the spread of ST410. The AdeABC efflux pump appeared to play an important role in the tigecycline resistance of A. nosocomialis.

Calcium channels are necessary for cardiac excitation–contraction (E–C) coupling, but Ca 2+ channel composition of fish hearts is still largely unknown. To this end, we determined transcript expression of Ca 2+ channels in the heart of zebrafish ( Danio rerio ), a popular model species. Altogether, 18 Ca 2+ channel α-subunit genes were expressed in both atrium and ventricle. Transcripts for 7 L-type (Ca v 1.1a, Ca v 1.1b, Ca v 1.2, Ca v 1.3a, Ca v 1.3b, Ca v 1.4a, Ca v 1.4b), 5 T-type (Ca v 3.1, Ca v 3.2a, Ca v 3.2b, Ca v 3.3a, Ca v 3.3b) and 6 P/Q-, N- and R-type (Ca v 2.1a, Ca v 2.1b, Ca v 2.2a, Ca v 2.2b, Ca v 2.3a, Ca v 2.3b) Ca 2+ channels were expressed. In the ventricle, T-type channels formed 54.9%, L-type channels 41.1% and P/Q-, N- and R-type channels 4.0% of the Ca 2+ channel transcripts. In the atrium, the relative expression of T-type and L-type Ca 2+ channel transcripts was 64.1% and 33.8%, respectively (others accounted for 2.1%). Thus, at the transcript level, T-type Ca 2+ channels are prevalent in zebrafish atrium and ventricle. At the functional level, peak densities of ventricular T-type ( I CaT ) and L-type ( I CaL ) Ca 2+ current were 6.3±0.8 and 7.7±0.8 pA pF −1 , respectively. I CaT mediated a sizeable sarcolemmal Ca 2+ influx into ventricular myocytes: the increment in total cellular Ca 2+ content via I CaT was 41.2±7.3 µmol l −1 , which was 31.7% of the combined Ca 2+ influx (129 µmol l −1 ) via I CaT and I CaL (88.5±20.5 µmol l −1 ). The diversity of expressed Ca 2+ channel genes in zebrafish heart is high, but dominated by the members of the T-type subfamily. The large ventricular I CaT is likely to play a significant role in E–C coupling.

*Co-First Authors Background: Currently, patients diagnosed with breast carcinoma undergo HER2 testing to direct clinical treatment decisions. At present, immunohistochemical (IHC) and in-situ hybridization methodologies are employed in the clinical setting to ascertain HER2 status. While these tests represent the current standard, their interpretation and variability of results with respect to HER2 prognostic and predictive value remains an outstanding issue. Aim: In this comparative study we assessed the utility of testing HER2 gene expression by RT-qPCR and the Luminex Quantigene® Plex 2.0 (Affymetrix) methodology against the clinically accepted IHC assay. Methods: Local cases from 2008-2010 that were clinically evaluated for HER2 were identified and underwent further pathologist review. In cases where there was sufficient tumor, formalin fixed paraffin embedded samples were retrieved. A total of 207 cases were identified which met selection criteria. Tumour sections were stained for HER2 and scored 0-3, following ASCO/CAP guidelines. For molecular assessment total RNA was extracted from tumours and those samples with sufficient RNA yield and quality were assessed for Her2 transcript level expression by RT-qPCR (n=129) and Luminex Quantigene® Plex 2.0 assays (n=166). Results: Results for RT-qPCR are relative to two normal breast calibrator samples and reported as the mean relative quantification (RQ) value. For HER2 IHC negative cases (0/1+), the mean score was 0.13 (0.004-1.84, SD ±0.22); equivocal cases (2+), mean score was 0.19 (0.007-0.72, SD ±0.18); and positive cases (3+), mean score was 1.51 (0.03-6.78, SD ±1.61). Student’s t-test was performed to compare the means between groups and results are as follows: negative vs. equivocal p=0.17; equivocal vs. positive p=0.0002; and negative vs. positive p Citation Format: Elizabeth N Kornaga, John B McIntyre, Alexander C Klimowicz, Natalia Guggisberg, Don G Morris, Anthony M Magliocco. Evaluation of RT-qPCR and luminex-based methodolgies in HER2 breast cancer testing [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P5-10-16.

To explore the factors for achieving early molecular responses (EMR; BCR-ABL1 ≤10% at 3 months, ≤1% at 6 months) by imatinib (IM), baseline characteristics including individual BCR-ABL1 transcript level, dose intensity, and IM trough level on day 29 were analyzed in 286 chronic phase chronic myeloid leukemia patients. Distinct predictive factors for achieving EMR at 3 months and 6 months were noted. Blast count at diagnosis and IM trough level on day 29 were significantly associated with an achievement of 3-month EMR. Early decline of BCR-ABL1 transcript, low Sokal risk, and mean daily dose (≥350mg/day) by 6 months were associated with an achievement of 6-month EMR. Understanding the predictive factors for EMR may provide additional information to guide clinical decisions on the changing therapies at each landmark.

Ixeris dentata var. albiflora is considered as a potential therapeutic agent against mithridatism, calculous, indigestion, pneumonia, hepatitis, and tumors as well as good seasoned vegetable in Far East countries. Phytoene synthase (PSY), phytoene desaturase (PDS) ξ-carotene desaturase (ZDS), lycopene β-cyclase (LCYB), lycopene ε-cyclase (LCYE), ε-ring carotene hydroxylase (CHXB), and zeaxanthin epoxidase (ZDS) are vital enzymes in the carotenoid biosynthesis pathway. We have examined these seven genes from I. dentata that are participated in carotenoid biosynthesis utilizing an Illumina/Solexa HiSeq 2000 platform. In silico analysis of the seven deduced amino acid sequences were revealed its closest homology with other Asteracea plants. Further, we explored transcript levels and carotenoid accumulation in various organs of I. dentata using quantitative real time PCR and high-performance liquid chromatography, respectively. The highest transcript levels were noticed in the leaf for all the genes while minimal levels were noticed in the root. The maximal carotenoid accumulation was also detected in the leaf. We proposed that these genes expressions are associated with the accumulation of carotenoids. Our findings may suggest the fundamental clues to unravel the molecular insights of carotenoid biosynthesis in various organs of I. dentata.

Previous studies showed that people in urban areas are possibly exposed to 60–110 dB of low frequency noise (LFN) defined as noise of ≤100 Hz in their daily life. Previous studies also showed increased health risks by exposure to high levels (130–140 dB) of LFN in animals. However, little is known about the health effects of exposure to an ordinary level of LFN. We biochemically and immunohistochemically assessed the effects of exposure to inaudible LFN for mice (12 h/day of 100 Hz LFN at 95 dB for 5 days), at a level to which people are possibly exposed in daily life, on a murine inner ear by targeting 9 stress-reactive molecules. There was more than a 5-fold increased transcript level of heat shock protein 70 (Hsp70) in the whole inner ear exposed to LFN. However, the transcript levels of the other 8 stress-reactive molecules including Hsp27 and Hsp90 were comparable in LFN-exposed and unexposed murine inner ears. Only the transcript level of Cebpβ among the previously reported 4 transcriptional activators for Hsp70 expression was more than 3-fold increased by LFN exposure. Hsp70 transcript expression levels in the inner ears 3 days after LFN exposure were comparable to those in unexposed inner ears. The protein level of Hsp70, but not the levels of Hsp27 and Hsp90, was also increased in the vestibule by LFN exposure. However, hearing levels as well as expression levels of Hsp70 protein in the cochleae were comparable in LFN-exposed mice and unexposed mice. Our results demonstrated that the inner ear might be one of the organs that is negatively affected by stress from inaudible LFN exposure. Moreover, LFN exposure might increase Hsp70 expression level via Cebpβ in the inner ear. Thus, Hsp70 and Cebpβ levels could be candidates of biomarkers for response to LFN exposure., ファイル公開：2019-06-01

Purpose Tyrosine kinase inhibitors (TKIs) are the cornerstones of treatment for patients with chronic myeloid leukaemia (CML). In recent years, several studies were conducted to evaluate the safety of TKIs discontinuation. We performed a systematic review of the literature to determine the incidence of CML relapse, to identify possible factors relapse rates and to evaluate the long-term safety in CML patients with stable undetectable BCR-ABL transcript level who discontinued TKIs. Design Studies evaluating TKIs discontinuation in CML patients with undetectable BCR-ABL transcript level were identified by electronic search of MEDLINE and EMBASE database until May 2015. Weighted mean proportion and 95% confidence intervals (CIs) of CML relapse was calculated using a fixed-effects and a random-effects model. Statistical heterogeneity was evaluated using the I 2 statistic. Results Fifteen cohort studies, for a total of 509 patients, were included. Nine studies were at low-risk of bias. All 15 studies included only patients on imatinib. Overall weighted mean molecular relapse rate of CML was 51% (95% CI 44–58%; I 2 = 55). Weighted mean molecular relapse rate at 6-month follow-up was 41% (95% CI 32–51%; I 2 = 78). Eighty percent of molecular relapses occurred in the first 6 months. All 509 patients were alive at 2-year follow-up and only one patient (0.8%, 95% CI 0.2–1.8%; I 2 = 0) has progressed to a blastic crisis. Conclusions Our findings suggest that imatinib discontinuation is feasible for the majority of CML patients with stable undetectable BCR-ABL transcript level. Approximately 50% of patients remain therapy-free after imatinib discontinuation. Restarting TKIs therapy was followed by a very high rate of molecular response, with no deaths 2 years after discontinuation.

Background Cell therapies have transformed the field of hematologic cancers, but treatment of solid tumours remains challenging. Emerging evidence suggests that T cell receptor (TCR)- engineered T cells targeting NY-ESO-1 hold promise for patients with solid tumours. NY-ESO-1 (CTAG1B) and LAGE1A (CTAG2) are tumour associated antigens (TAA) that share the SLLMWITQC peptide bound to human leukocyte antigen HLA-A*02 and are expressed in various cancers. GSK3377794 are autologous T cells engineered to express an affinity enhanced NY-ESO-1c259 TCR recognizing the SLLMWITQC/HLA-A*02 complex. Cell therapy trials using NY-ESO-1c259 TCR have shown promising efficacy in patients with synovial sarcoma, metastatic melanoma and multiple myeloma. Recently, a separate study using T cells targeted to NY-ESO-1 resulted in a partial response in a patient with advanced lung adenocarcinoma. This study aims to assess: 1) the prevalence of NY-ESO-1 and LAGE1A in various malignancies 2) GSK3377794 function against various tumours and 3) means to selectively modulate TAA expression in tumours to increase potential patient benefit. Methods Gene and transcript level RNAseq data for NY-ESO-1 and LAGE1A respectively, were extracted from TCGA B38 using ArrayStudio v10. A solid tumour biobank was established. HLA-A*02 expression was confirmed by flow cytometry and NY-ESO-1/LAGE1A levels were measured via qRT-PCR. Epigenetic modifiers were applied to increase TAA expression in tumours in vitro and in vivo. GSK3377794 anti-tumour function was determined by IFN-γ secretion. Results NY-ESO-1 and LAGE1A expression varied across malignancies and disease stages. Prevalence is high in synovial sarcoma and multiple myeloma (60-70%) and lower in lung cancer (12-15%). IFN-γ production by GSK3377794 positively correlates with antigen expression. Treatment with epigenetic modifiers lead to increased NY-ESO-1/LAGE1A expression in nonimmunogenic lung tumours, enhancing specific targeting by GSK3377794. Conclusions To date this is the most comprehensive analysis, using a set of clinically relevant sample specimens, that correlates antigen expression levels with functional responses of GSK3377794 in solid tumors. Epigenetic modifiers can improve GSK3377794 anti-tumour effect in lung cancer. Legal entity responsible for the study GlaxoSmithKline. Funding GlaxoSmithKline. Disclosure I. Eleftheriadou: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. S. Brett: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. A. Domogala: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. L. Patasic: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. M.A. Kijewska: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. K. Soor: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. M. Georgouli: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. J. Dopierala: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. P. Fisher: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. J. Jing: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. J. Euesden: Full / Part-time employment: GlaxosmithKline. K.R. Auger: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. R. Roberts: Full / Part-time employment: GlaxosmithKline. S. O’Sullivan: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. L. Castelletti: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. M. Damm: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. D. Pankov: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. L.A. Johnson: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. A. Shalabi: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline. C. Britten: Shareholder / Stockholder / Stock options, Full / Part-time employment: GlaxosmithKline.

RT-qPCR is the standard method for studying changes in relative transcript level in different experimental and clinical conditions and in different tissues. No validated reference genes have been reported for the normalization of transcript level in platelets. The very low level of platelet RNA and the elimination of leukocyte contamination represented special methodological difficulties. Our aims were to apply a simple technique to separate platelets for transcript level studies, and select the most stable reference genes for platelets from healthy individuals and from patients with the history of myocardial infarction. We developed a simple, straightforward method of platelet separation for RNA isolation. Platelet activation was inhibited by using acid-citrate-dextrose for anticoagulation and by prostaglandin E1. Leukocyte contamination was eliminated by three consecutive centrifugations. Samples prepared by this method were free of leukocytes, showed no inhibition in PCR reaction and no RNA degradation. The assay demands low blood volume, which complies with the requirements of everyday laboratory routine. Seventeen potential reference genes were investigated, but eight of them were excluded during optimization. The stability of the remaining genes, EEF2, EAR, ACTB, GAPDH, ANAPC5, OAZ1, HDGF, GNAS, and CFL1, were determined by four different descriptive statistics. GAPDH, GNAS, and ACTB were shown to be the most stable genes in platelets of healthy individuals, while HDGF, GNAS, and ACTB were the most stable in platelets of patients with the history of myocardial infarction. The results confirm that data normalization needs assessment of appropriate reference genes for a particular sample set.

© 2016 The Japanese Society for Regenerative Medicine. RNA therapeutics mainly control gene expression at the transcript level. In contrast to conventional gene therapy which solely increases production of a protein, delivered RNAs can enhance, reduce or abolish synthesis of a particular protein, which control its relevant activities in a more diverse fashion. Thus, they hold promise to treat many human diseases including myocardial infarction (MI). MI is a serious health burden that causes substantial morbidity and mortality. An unmet clinical need for treating MI is the recovery of cardiac function, which requires regeneration of the functional tissues including the vasculature, nerves, and myocardium. Several classes of RNA therapeutics have been investigated in preclinical MI models, and the results have demonstrated their benefits and encourage their future development. In this review, we summarize the common RNA therapeutic approaches and highlight their application in MI therapy.

Identification of prognostic factors and risk-based post-remission therapy was proposed to improve the outcomes of acute myeloid leukemia (AML) and a mutation of FLT3 has been reported to be a risk factor, especially for pediatric patients. Recently, FLT3 expression level was implicated to have prognostic significance in adults, but little is known for childhood AML. To define the prognostic significance, transcript level of FLT3 was analyzed in 52 pediatric AML patients. The median copy number of FLT3 was 4.6 x 10 3 (40-5.9 × 10 7 copies)/1.0 x 10 6 GAPDH copy, and the relapse free survival of patients with high transcript level of FLT3 (>10 6 copy number) (0%) was significantly lower than that of the others (53.2%). High transcript level of FLT3 was associated with a markedly high risk of relapse. The development of new therapeutic scheme such as a frontline allogeneic stem cell transplantation or administration of FLT3 inhibitor is needed to improve outcomes.