M1 Cases Research Articles

First Description Of a t(10;11)(q22;q23)/MLL-TET1 Reciprocal Translocation In a T Lymphoblastic Lymphoma With Subsequent Lineage Switch To Acute Myelomonocytic Myeloid Leukemia

TET1 genomic breakpoints and clinical features of MLL-TET1 rearrangements have been described in 13 acute leukemia cases, 11 in AML, 2 in B cell-precursor ALL. The incidence of this rare translocation was evaluated to 0.3% of MLL rearranged AML cases (5 out of 1590 MLL [Meyer C. Leukemia 2013]). Although those cases are very uncommon, their study can improve our current understanding of leukemogenesis. We report here the first t(10;11) MLL-TET1 positive case of lymphoblastic T lymphoma occurring in a 31 years old male patient, with a subsequent transformation to AML.The patient was referred for a large mediastinal mass and right pleural effusion. Mediastinal and bronchus biopsies led to the diagnosis of a precursor-T cell lymphoblastic lymphoma, expressing CD3, CD5, CD4, CD8, CD10 antigens, without any expression of CD34 or CD79.Molecular analyses of the malignant T-cells showed a clonal TCR gamma-chain gene rearrangement together with HOXA10 overexpression. FISH analysis showed a MLL breakage. The partner gene, TET1, was identified using RP11-9E13 and RP11-314J18 BACs, corresponding to the recurrent translocation t(10;11)(q22;q23). MLL-TET1 fusion transcript was detected (intron 8 of TET1 fused to exon 8 of MLL), as well as its reciprocal transcript.The patient was treated following the (GELA) LL03 protocol, and was considered in complete remission after induction and consolidation phases. Fourteen months after the diagnosis, a bone marrow examination was performed for thrombopenia (6 G/L) which revealed a myelomonocytic acute leukemia with trilineage dysplasia.At this time, The MLL-TET1 fusion transcript as well as HOXA10 overexpression was still present but the TCR rearrangement was not detected. A non-familial donor allogeneic bone marrow transplant was performed in CR after intensive chemotherapy, that was complicated by a grade IV acute graft-versus-host disease. The patient died 54 days after the transplant of bacterial sepsis leading to multi-organ failure.MLL-TET1 fusions have been described in 13 cases in the literature, mainly in AMLs (11/13 cases, mostly AML M4 or M5 cases) and in B-ALLs (2/13 cases). The case reported here presents two interesting features. Firstly, this patient harbors the first MLL-TET1 fusion reported to date in T-ALL, and secondly this case presented a lymphoid to myeloid phenotype switch during the time course of the disease.This strongly suggests that the translocation occurred very early during hematopoietic differentiation, prior to the lymphoid/myeloid commitment. As described for the Ph1 chromosome rearrangement, the t(10;11) translocation may arise in hematopoietic stem cell rather than in committed progenitors. These features are also described in 8p11 stem cell syndrome that involves FGFR1. In both cases, genetic rearrangements arise in myeloid or lymphoid neoplasms with possible subsequent transformation.TET family enzymes convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) and play a key role in active DNA demethylation. TET1 and TET2 are also the key enzymes responsible for the presence of 5hmC in mouse embryonic stem cells (ESCs) (Koh KP., Cell Stem Cell 2011), and TET1 functions to regulate the lineage differentiation potential of ESCs. In addition to its role in DNA demethylation, TET1 interacts physically with NANOG and NANOG/TET1 co-occupy genomic loci of genes associated with both maintenance of pluripotency and lineage commitment in embryonic stem cells (Costa Y., Nature 2013). Taken together, these observations enlighten possible mechanism of the lineage switch observed in this case, and may be a rationale for using demethylating agents in MLL-TET1 neoplasms. Disclosures:No relevant conflicts of interest to declare.

Homoharringtonine In Combination With Cytarabine and Etoposide For Induction Therapy In Patients With De Novo Acute Myelogenous Leukemia

Homoharringtonine (HHT) is one of antineoplastic alkaloid extracted from Cephalotaxus of China. Its main function is to inhibit protein synthesis. Its cytotoxicity is cell cycle specificity, which mainly affects G1 and G2-cyle cells and can induce the apoptosis of acute myeloid leukemic cells and has no obvious cross resistance to the drugs like daunorubicin and cytarabine (Ara-C).This research has made a preliminary assessment to the efficacy and safety of HAE regimen's induction therapy to patients with de novo acute myeloid leukemia (AML). From March 2011 to June 2013, 35 patients with de novo AML have been enrolled, of which 18 are males and 17 females with median age as 42y (19∼60); there are three M0 and M1 cases respectively, thirteen M2 cases, six M4 cases and ten M5 cases. The myeloid leukemia cell is 71.5% (20.5%∼94%). According to the detection result of chromosome and gene mutation, the patients are divided into eight high-risk cases and 27 low-and-medium-risk cases. The chemotherapy program is HAE, HHT 2.5mg/m2/d, d1–7, Ara-C 100mg/m2/d, d1–7, Etoposide 60mg/m2/d, d1–7. If complete remission (CR) or CR with no recovery of hemogram (CRi) is obtained after one course of treatment, another one course of treatment will be given as consolidation therapy. If partial remission (PR) is obtained, another one course of treatment will be made and if CR or CRi is obtained then, another one course of treatment will be made for consolidation therapy, otherwise, exit the test. ResultCurative effect of one course of treatment, 23 (65.7%) cases obtaining CR or CRi and 4 (11.4%) cases obtaining PR, of which 3 cases obtained CR or CRi after another one course of treatment. The total effective rate of 2 courses of treatment was 74.3% (26/35), 6 (17.1%) cases to which the treatment achieved no effect (including 1 case obtaining PR in the first course of treatment), 3 (8.6%) cases encountered earlier death. The time of median follow-up is 11.2m (1.1∼28.7m). In the 26 patients achieved CR or CRi, 4 cases encounter recurrent of disease (15.4%), with the median recurrence free survival (RFS) not met, and the 2-years estimated RFS was 73%. In the 35 patients, 5 (14.3%) cases died, including 3 cases of earlier death and 2 cases die of recurrence of disease, with the median overall survival (OS) not met, the 2-years estimated OS was 83%. In the 26 patients obtaining CR or CRi, the average RFS without recurrence of low-and-medium-risk and high-risk patients was 24.9 ± 2.0m and 14.5 ± 0.4m (p = 0.465) respectively, and in the 35 patients, the average OS of low-and-medium-risk and high-risk patients was 25.5 ± 1.8m and 11.6 ± 3.7m (p = 0.236) respectively. Non-hematologic adverse reaction of HAE program includes infection (88.6%, 31/35), diarrhea (11.4%, 4/35), pulmonary fungal infection (20.0%, 7/35) and atrial fibrillation (2.9%, 1/35), of which grade 3∼4 adverse reaction is infection (37.1%, 13/35). ConclusionHAE program is tolerable and effective program for first-line treatment of AML. Disclosures:No relevant conflicts of interest to declare.

On the structure of positive maps. II. Low dimensional matrix algebras

We use a new idea that emerged in the examination of exposed positive maps between matrix algebras to investigate in more detail the differences and similarities between unital positive maps on \documentclass[12pt]{minimal}\begin{document}$M_{2} (\mathbbm {C})$\end{document}M2(C) and \documentclass[12pt]{minimal}\begin{document}$M_{3}(\mathbbm {C})$\end{document}M3(C). Our main tool stems from classical Grothendieck theorem on tensor product of Banach spaces and is an older and more general version of Choi-Jamiołkowski isomorphism between positive maps and block positive Choi matrices. It takes into account the correct topology on the latter set that is induced by the uniform topology on positive maps. In this setting, we show that in \documentclass[12pt]{minimal}\begin{document}$M_{2}(\mathbbm {C})$\end{document}M2(C) case a large class of nice positive maps can be generated from the small set of maps represented by self-adjoint unitaries, 2Px with x maximally entangled vector and \documentclass[12pt]{minimal}\begin{document}$p\otimes \mathbb {1}$\end{document}p⊗1 with p rank 1 projector. We indicate problems with passing this result to \documentclass[12pt]{minimal}\begin{document}$M_{3}(\mathbbm {C})$\end{document}M3(C) case. Among similarities, in both \documentclass[12pt]{minimal}\begin{document}$M_{2}(\mathbbm {C})$\end{document}M2(C) and \documentclass[12pt]{minimal}\begin{document}$M_{3}(\mathbbm {C})$\end{document}M3(C) cases any unital positive map represented by self-adjoint unitary is unitarily equivalent to the transposition map. Consequently, we obtain a large family of exposed maps. Furthermore, for \documentclass[12pt]{minimal}\begin{document}$M_{3}(\mathbbm {C})$\end{document}M3(C) there appear new non-trivial class of maps represented by Choi matrices with square equal to a projector. We examine this case. We also investigate a convex structure of the Choi map, the first example of non-decomposable map. As a result the nature of the Choi map will be explained.

The Overall Prevalence of Metastasis in T1 Esophageal Squamous Cell Carcinoma

T1 esophageal squamous cell carcinoma (ESCC) has a low, but still present, risk of lymph node (LN) metastasis. Endoscopic mucosal resection (EMR) or endoscopic submucosal dissection (ESD) is often applied for T1 ESCC. To achieve successful treatment by EMR/ESD, the risk of LN metastases, LN recurrence, and hematological recurrence need to be better understood. The aim of this study was to determine the precise risk for metastasis in T1 ESCC. We divided 295 patients with T1 ESCC who underwent surgery and/or ESD/EMR into 6 categories (m1, m2, m3, sm1, sm2, and sm3). Their risks of LN metastasis, LN recurrence, hematological recurrence, and the outcome were determined. The rates of LN metastasis and LN recurrence were 0% in m1 and m2, 9% in m3, 16% in sm1, 35% in sm2, and 62% in sm3 cases. The incidence of hematological recurrence was 0% in m1, m2, m3, and sm1 cases; 9% in sm2 cases; and 13% in sm3 cases. The overall risk of metastasis was 9% in m3, 16% in sm1, 38% in sm2, and 64% in sm3 patients. The 5-year disease-specific survival rates were 100% in m1, m2, and m3; 90.9% in sm1; 78.8% in sm2; and 68.6% in sm3 patients. Statistically, both lymphatic and venous invasion were selected as predictive markers for metastasis. In m3 patients, positivity for either of these had an odds ratio for metastasis of 7.333 (P = 0.093). Our study provides a precise assessment of the comprehensive risk of metastasis and feasible predictive markers for T1 ESCC.

In Patients with Colorectal Cancer, Preoperative Serum Interleukin-6 Level and Granulocyte/Lymphocyte Ratio Are Clinically Relevant Biomarkers of Long-Term Cancer Progression

Objective: To investigate the clinical significance of the serum inflammatory cytokines and the blood granulocytes/lymphocytes (G/L) ratio in patients with colorectal cancer. Methods: Forty-six patients underwent surgery for colorectal cancer. The G/L ratio and serum inflammatory cytokines including interleukin (IL)-1β, IL-6, IL-8, tumour necrosis factor-α (TNF-α), granulocyte colony-stimulating factor and macrophage colony-stimulating factor (M-CSF) were measured before surgery and correlated with the Tumour Node Metastasis classification and overall survival. Results: Serum IL-6 in T3-4, N1-2, M1 cases and cancer stages III-IV sub-groups was significantly higher than in Tis-2, N0, M0 and cancer stages 0-II sub-groups. The G/L ratio, serum M-CSF and TNF-α in T3-4 cases were significantly higher than in Tis-2 cases. Furthermore, the G/L ratio and serum IL-6 showed a significant inverse correlation with the overall survival, while the G/L ratio showed a significant positive correlation with serum IL-6, TNF-α and M-CSF. Multivariate analysis showed that the serum IL-6 level and G/L ratio were independent risk factors for poor prognosis. Conclusions: In this investigation, a pre-operative high level of serum IL-6 and the G/L ratio appeared to be significant predictive factors for cancer progression and poor prognosis. Accordingly, these variables might be clinically relevant biomarkers in patients with colorectal cancer.

Clinical and molecular biologic characteristics of childhood acute leukemia with mixed lineage leukemia gene rearrangement

Objective To analyze the frequency of mixed lineage leukemia (MLL) gene rearrangement, the frequent types of fusion genes and clinical characteristics of childhood acute leukemia (AL) with MLL gene rearrangement. Methods Morphological and molecular characteristics of 87 AL patients with MLL gene rearrangement were studied and analyzed.MLL fusion gene was detected by way of reverse transcription polymerase chain reaction (RT-PCR). Results Eighty-seven cases with MLL gene rearrangement were found in 1209 AL patients with incidence of 6.41% and 9.36% respectively in ALL and in acute myelocytic leukemia (AML) respectively.Fifty-eight cases of ALL were all B-ALL, 28 cases of AML included 17 cases of M5, 5 cases of M4, 4 cases of M2, 1 case of M3 and 1 case of M6.While there was 1 case of mixed of lineage leukemia and myeloid and T-lymphoblastic antigen presentation.The clonal chromosomal aberration was detected in 45 out of 76 cases(59.21%), and chromosome 11q23 aberration were observed in 28 cases(36.84%). There were 7 different kinds of fusion genes, including MLL-AF9 in 25 cases, dupMLL in 25 cases, MLL-AF4 in 17 cases, MLL-AF10 in 9 cases, MLL-ENL in 8 cases, MLL-AF1q in 2 cases, and MLL-AF6 in 1 case.Among the cases of MLL-AF4, MLL-AF9, MLL-AF10, MLL-ENL and dupMLL, there were statistical differences in lineage, age and initial white blood cell count (WBC) (all P<0.05). Conclusions In childhood AL with MLL gene rearrangement, B-ALL is more common in ALL, whereas M5 and M4 are more common in AML.The common types of fusion genes are dupMLL, MLL-AF9 and MLL-AF4.Patients with the different kinds of MLL fusion gene may present different clinical characteristics. The most common ALL cases are those with MLL/AF4 and MLL/ENL who may be younger with higher WBC than the others. Key words: Leukemia, acute; Mixed lingeage leukemia; Fusion gene; Nested reverse transcription polymerase chain reaction; Child

Preoperative Serum Lactate Dehydrogenase Levels in Colorectal and Gastric Cancer: A Hospital-Based Case–Control Study

To reveal correlations between serum lactate dehydrogenase (LDH) levels and various clinicopathological parameters in colorectal and gastric cancer. A hospital-based case-control study was conducted by measuring serum LDH levels in 140 patients with colorectal cancer, 40 patients with gastric cancer and 20 hospital controls. LDH levels did not differ between patients with colorectal or gastric cancer and hospital controls. In colorectal cancer, LDH values were significantly higher in T4, N2 and/or M1 cases, with high specificities and negative predictive values, but low sensitivities and positive predictive values. On the other hand, there were no significant associations in gastric cancer. Serum LDH levels are increased in T4, N2 and/or M1 colorectal cancer. Most patients with early-stage cancer have normal LDH values, whereas in advanced stages, some patients have increased LDH values and others have normal values.

The horizontal branch in the UV colour–magnitude diagrams – II. The case of M3, M13 and M79†

We present a detailed comparison between far-UV/optical colour Magnitude Diagrams obtained with high-resolution Hubble Space Telescope data and suitable theoretical models for three Galactic Globular Clusters: M3, M13 and M79. These systems represents a classical example of clusters in the intermediate metallicity regime that, even sharing similar metal content and age, show remarkably different Horizontal Branch morphologies. As a consequence, the observed differences in the colour distributions of Horizontal Branch stars cannot be interpreted in terms of either first (metallicity) or a second parameter such as age. We investigate here the possible role of variations of initial Helium abundance (Y). Thanks to the use of a proper setup of far-UV filters, we are able to put strong constraints on the maximum Y (Y_{max}) values compatible with the data. We find differences Delta Y_{max} ~ 0.02-0.04 between the clusters with M13 showing the largest value (Y_{max} ~ 0.30) and M3 the smallest (Y_{max} ~ 0.27). In general we observe that these values are correlated with the colour extensions of their Horizontal Branches and with the range of the observed Na-O anti-correlations.

Associations of adiponectin and leptin with stage and grade of PSA-detected prostate cancer: the ProtecT study

Obesity has been associated with an increased risk of advanced and fatal prostate cancer; adipokines may mediate this association. We examined associations of the adipokines leptin and adiponectin with the stage and grade of PSA-detected prostate cancer. We conducted a nested case-control study comparing 311 men with mainly locally advanced (≥T3, N1, or M1 cases) vs. 413 men with localized (T ≤2 & NX-0 & M0 controls) PSA-detected prostate cancer, recruited 2001-2009 from 9 UK regions to the ProtecT study. Associations of body mass index and adipokine levels with prostate cancer stage were determined by conditional logistic regression and with grade (Gleason score ≥7 vs. ≤6) by unconditional logistic regression. Adiponectin was inversely associated with prostate cancer stage in overweight and obese men (OR 0.62; 95 % CI 0.42-0.90; p = 0.01), but not in normal weight men (OR 1.48; 0.77-2.82; p = 0.24) (p for interaction 0.007), or all men (OR 0.86; 0.66-1.11; p = 0.24). There was no compelling evidence of associations between leptin or leptin to adiponectin ratio and prostate cancer stage. No strong associations of adiponectin, leptin, or leptin:adiponectin ratio with grade were seen. This study provides some evidence that adiponectin levels may be associated with prostate cancer stage, dependent on the degree of adiposity of the man. Our results are consistent with adiponectin countering the adverse effects of obesity on prostate cancer progression.

Comparable Toxicity and Outcome of Acute Myeloid Leukemia (AML) Fit Patients Therapy Independently If Daunorubicin or Idarubicin Were Added to Cytarabine/Cladribine Induction Regimens – the Single Centre Study

AbstractAbstract 4337This trial is a continuation of the earlier studies concerning optimalization of induction chemotherapy in acute myeloid (AML) patients using purine analogues (Ho≥owiecki J., Grosicki S., Robak T. et al. Leukemia 2004, Holowiecki J, Grosicki S, Giebel S. et al. J Clin Oncol. 2012).The goal of the study was to evaluate the toxicity and efficacy of combination including idarubicin – IAC (idarubicin 10 mg/m2/d iv, d 1–3; cytarabine 200 mg/m2/d ci, d 1–7, cladribine 5 mg/m2/d d 1–5) in comparison with DAC (daunorubicin 60 mg/m2/d iv d 1–3 in place of idarubicin).The primary endpoints were complete remission rate (CR) and toxicity, the secondary one was overall survival (OS).Patients who achieved CR received two courses of subsequent intensive consolidation: HAM (HD AraC, mitoxantrone) and HD AraC. Reduction of consolidation was accepted in patients treated with early alloBMT. In case of partial remission (PR) after the first induction course the same regimen was repeated. Patients with no remission (NR) or with PR after 2 induction courses were withdrawn from the study.Between 03.2009 and 03.2012, 52 adult untreated patients aged median 57 years (20–72), treated in one center - Department of Hematology of City Hospital in Chorzow/Poland were included to this study; 46% of study participants were male, 54% female and they were evaluated as fit with possibility of intensive chemotherapy including HSCT. PML/RAR alfa positive – FAB M3 cases were excluded.DAC regimen was a standard induction in this group of AML patients. When there was no possibility to apply DAC regimen because of not having access to daunorubicin in our center, the IAC regimen was used.Material and the results are summarized in the table.OutcomeDAC (n = 30)IAC (n = 22)pAge (median)20–72 (55)38–70 (59)NSSex (%)K-53, M-47K-55, M-45NSWBC at dgn. × 103/L median (range)27,7 (2,3–257)11,0 (0,9–210)NSCytogenetic risk (%)Low00NSintermediate5064NSHigh75NSNo data4331NSHSCT (n) auto/allo2/71/5NSCR rate (%)7059NS3-yr OS (%)2623NS2-yr LFS (%)2836NSThe study groups were well balanced with respect to age, sex, WBC and cytogenetics. The entire CR rates in both groups did not differ, and they were similar to previously published results after DAC induction regimen. All patients developed WHO grade IV thrombocytopenia and agranulocytosis. The frequency and severity of infections, mucositis, vomiting, diarrhea, alopecia, polyneuropathy as well as of cardiac, liver or kidney dysfunctions were comparable in particular arms. There were no significant differences in the OS and LFS rates.In conclusion, the results of our study authorize the replacement of daunorubicin to idarubicin in DAC induction regimen without risk of additional toxicity, and with similar outcome. Disclosures:No relevant conflicts of interest to declare.

Dysregulation of the Imprinted DLK1-DIO3 Locus in Promyelocytic Leukemia

Abstract 3500Small non-coding RNAs (sncRNAs) have emerged as important regulators of multiple cellular processes, with a growing body of evidence suggesting their dysregulation and possible pathogenic role in many malignancies, including acute myeloid leukemia (AML). Previous high throughput sncRNA transcriptome sequencing studies have focused on the 15–30 nucleotide fraction that primarily consists of microRNAs (miRNAs), thus excluding many larger, potentially important sncRNAs, such as small-nucleolar RNAs (snoRNAs) and piwiRNAs. To address this “sequencing gap”, we extended our transcriptome sequencing to include RNA species of 15–75 nucleotides in length. Here, we report the small RNA transcriptome sequencing of 28 cases of AML. As a control, we also sequenced the small RNA transcriptome in CD34+ progenitors (n = 4) and promyelocytes (n=2) from healthy donors. The most common class of sncRNA detected in all samples was small-nuclear/snoRNAs (∼40% of reads), followed by miRNAs (∼17% of reads), ribosomal/transfer RNAs (15% of reads), unannotated/miscellaneous sncRNAs (∼15% of reads), and piwiRNAs (∼10% of reads). Partek Genomics Suite was used to identify significantly dysregulated sncRNAs using a p-value<0.05, false discovery rate of 5%, and a minimum 2 fold-change from normal CD34+ cells. Compared to normal CD34+ cells, expression of 158 sncRNAs was significantly increased and 29 were significantly decreased in de novo AML samples.The most striking example of dysregulation was observed in acute promyelocytic leukemia (M3 AML). The DLK1-DIO3 locus at 14q32.2 contains 41 snoRNA genes belonging to the SNORD112–114 family, in addition to a large number of miRNAs. Compared with normal CD34+ cells or non-M3 AML, 31 of the 41 snoRNAs in this locus were massively upregulated (6–2,000 fold) in M3 AML samples. Concordantly large increases (10–1,000 fold) of 20 different miRNAs in this locus were also observed in M3 AML. We extended these findings to an independent cohort of M3 and non-M3 AML cases (n = 187) for which RNA transcriptome sequencing was available (RJW and TJL, personal communication, on behalf of the TCGA AML analysis group). Analysis of this data confirmed massive up-regulation of sncRNAs in the DLK1-DIO3 locus that was restricted to M3 AML. We considered the possibility that the increase in sncRNAs in this locus was secondary to a promyelocyte-specific expression pattern in M3 AML cells, but expression of sncRNAs in normal promyelocytes was lower than that observed in normal CD34+ cells, making this hypothesis unlikely.The DLK1-DIO3 locus is one of the best characterized imprinted regions in the human genome. The paternally-derived protein coding genes in this locus (DLK1, RTL1 and DIO3) showed no dysregulation in M3-AML in contrast to the aberrantly expressed, maternally-derived sncRNAs. While the function of the sncRNAs in this region is largely unknown, there is evidence they may contribute to stem cell pluripotency (Stadtfeld et al. Nature 2010, Liu et al. JBC 2010). Since imprinting at this locus is controlled by methylation of several differentially methylated regions (DMRs), we analyzed array-based methylation data from the TCGA cohort to determine if imprinting was disrupted. No difference in methylation status of the DMRs was observed between M3 and non-M3 AML. Moreover, based on expressed germline single nucleotide polymorphisms, mono-allelic expression of sncRNAs in this locus was preserved. Together, these data show that imprinting of the DKL1-DIO3 locus is not disrupted in M3-AML samples, and that dysregulation of the sncRNAs in this region occur through an imprinting-independent mechanism. In summary, extended small RNA transcriptome sequencing is a valuable new tool to analyze malignant cells; it successfully identified massive dysregulation of sncRNAs in the DKL1-DIO3 locus in M3 AML. The contribution of the aberrantly expressed sncRNAs within this locus to the pathogenesis of M3 AML will require additional study. Disclosures:No relevant conflicts of interest to declare.

Study on low-energy sputtering near the threshold energy by molecular dynamics simulations

Using molecular dynamics simulation, we have studied the low-energy sputtering at the energies near the sputtering threshold. Different projectile-target combinations of noble metal atoms (Cu, Ag, Au, Ni, Pd, and Pt) are simulated in the range of incident energy from 0.1 to 200 eV. It is found that the threshold energies for sputtering are different for the cases of M1 &amp;lt; M2 and M1 ≥ M2, where M1 and M2 are atomic mass of projectile and target atoms, respectively. The sputtering yields are found to have a linear dependence on the reduced incident energy, but the dependence behaviors are different for the both cases. The two new formulas are suggested to describe the energy dependences of the both cases by fitting the simulation results with the determined threshold energies. With the study on the energy dependences of sticking probabilities and traces of the projectiles and recoils, we propose two different mechanisms to describe the sputtering behavior of low-energy atoms near the threshold energy for the cases of M1 &amp;lt; M2 and M1 ≥ M2, respectively.

Personalized cancer therapy for stage IV non-small cell lung cancer: Combined use of active hexose correlated compound and genistein concentrated polysaccharide

Abstract This study aims to provide a strategy for personalized cancer therapy that results in increased survival time with better and more stable quality of life (QOL). Methods and patients Active hexose correlated compound (AHCC) and genistein concentrated polysaccharide (GCP) were administered to patients diagnosed with stage IV non-small cell lung cancer (NSCLC) with distant metastasis (M1). AHCC and GCP were consumed orally by concurrent or sequential use. Long-term follow-up study was conducted from January 1996 to January 2012 using the individualized evidence-based medicine (EBM) approach. Forty-two cases of NSCLC were enrolled in this study. Results The cumulative survival rate after 1, 2, 3, 4, and 5 years of treatment were 80.6%, 56.8%, 24.3%, 8.1%, and 5.4%, respectively. Median survival time (MST) was relatively longer at 28.26 months, as compared to that of current cancer therapies that solely involve chemotherapy. Evaluation of QOL showed that 40% of the 42 M1 cases achieved excellent (Class A) and good (Class B) QOL according to our 5-level QOL scoring. Conclusions AHCC and GCP resulted in increased patient survival time, better QOL, and reduced adverse effects related to cancer chemotherapy.

Expression of coagulation factor XIII subunit A in acute promyelocytic leukemia

Leukemic cells often express markers, which are not characteristic of their particular cell lineage. In this study, we identified the "A" subunit of coagulation factor XIII (FXIII-A) in leukemic promyelocytes in de novo AML M3 cases. The cytoplasmic presence of factor XIII-A has previously been shown only in platelets/megakaryocytes and monocytes/macrophages. Furthermore, more recently we described the presence of FXIII-A in leukemic lymphoblasts. We studied 14 patients with this rare type of acute leukemia in a period of 4 years and investigated their bone marrow samples by 3-color flow cytometry upon diagnosis, mainly focusing on FXIII-A expression of leukemic cells. We detected FXIII-A also by ELISA, Western-blot, and confocal laser scanning microscopy. This was a homogenous group of AML M3 patients with translocation t(15;17)(q22;q21) detected by fluorescence in situ hybridization (FISH). In 10 out of 14 samples, FXIII-A was detectable by flow cytometry and was coexpressed with markers characteristic for leukemic promyleocytes (CD45dim/CD13+/CD33+/CD117+/cyMPO+ and HLA-DR-/CD34-/CD14-/CD15-). Staining for the markers GPIIb and GPIX were negative, and FXIII-A was identified in the cytoplasm of the cells by confocal microscopy in a relatively high quantity, as measured by ELISA. By Western blot analysis we could identify FXIII-A in the native 82 kDa form and in cleaved forms corresponding to cleavage products observed when purified FXIII-A was treated by human neutrophil elastase. This novel expression site of FXIII-A in AML M3 can be considered as a leukemia associated immunophenotype and may have pathophysiological significance.

Immunophenotypic characterization of acute leukemia at a public oncology reference center in Maranhão, northeastern Brazil.

The incidence of acute leukemia (AL) subtypes varies according to geographical distribution. The aim here was to determine the incidence of morphological and immunophenotypic AL subtypes in the state of Maranhão, Brazil, and to correlate the expression of aberrant phenotypes in children with acute lymphoblastic leukemia (ALL) with prognostic factors. Single prospective cohort study at a public oncology reference center in Maranhão. Seventy AL cases were diagnosed between September 2008 and January 2010. For the diagnosis, complete blood cell counts, myelograms (at diagnosis and at the end of the induction phase), cytochemical analysis and immunophenotyping were performed. Among adult patients (n = 22), the incidence of AL types was: ALL (22.7%) and acute myeloid leukemia (AML) (77.3%). The subtype AML M0 occurred most frequently (29.4%). In children (n = 48), the types were: AML (18.7%), most frequently subtype AML M4 (33.4%); biphenotypic acute leukemia (BAL) (4.2%); and ALL (77.1%), including the subtypes B-ALL (72.9%) and T-ALL (27.1%). Among the children with ALL, there were no statistically significant differences between patients with and without aberrant phenotypes, in relation to hematological parameters and treatment response. This work demonstrates that the frequencies of AML M0 cases among adults and T-ALL cases among children in Maranhão were high. This suggests that there may be differences in AML subtype incidence, as seen with ALL subtypes, in different regions of Brazil. No association was found between the expression of aberrant phenotypes and prognostic factors, in children with ALL.

Prostate cancer in the elderly: frequency of advanced disease at presentation and disease-specific mortality

BACKGROUND The objectives of this study were to determine the frequency of metastatic (M1) prostate cancer (PC) at presentation in different age groups, to examine the association of age with PC-specific mortality, and to calculate the relative contribution of different age groups to the pool of M1 cases and PC deaths. METHODS Records from 464,918 patients who were diagnosed with PC from 1998 to 2007 were obtained from the Surveillance, Epidemiology, and End Results (SEER) database. The patients were categorized according to age into groups ages <50 years, 50 to 54 years, 55 to 59 years, 60 to 64 years, 65 to 69 years, 70 to 74 years, 75 to 79 years, 80 to 84 years, 85 to 89 years, and ≥ 90 years. The cumulative incidence of death from PC was computed using the Gray method. RESULTS The frequency of M1 PC at presentation was 3% for the group aged <75 years, 5% for the group ages 75 to 79 years, 8% for the group ages 80 to 84 years, 13% for the group ages 85 to 89 years, and 17% for the group aged ≥ 90 years. The 5-year cumulative incidence of death from PC was 3% to 4% for all patients with PC in any category aged <75 years, 7% for patients ages 75 to 79 years, 13% for patients ages 80 to 84 years, 20% for patients ages 85 to 89 years, and 30% for patients aged ≥ 90 years. Although patients aged ≥ 75 years at PC diagnosis represented just over a quarter (26%) of all PC cases, they contributed almost half (48%) of all M1 cases and more than half (53%) of all PC deaths. CONCLUSIONS Compared with younger patients (aged <75 years), older patients were more likely to present with very advanced disease, had a greater risk of death from PC despite higher death rates from competing causes, and contributed more than half of all PC deaths. Awareness of this issue may improve future outcomes for elderly patients with PC.

Complete Sequencing and Comparison of 12 Normal Karyotype M1 AML Genomes with 12 t(15;17) Positive M3-APL Genomes

Abstract 404To characterize the genomic events associated with distinct subtypes of AML, we used whole genome sequencing to compare 24 tumor/normal sample pairs from patients with normal karyotype (NK) M1-AML (12 cases) and t(15;17)-positive M3-AML (12 cases). All single nucleotide variants (SNVs), small insertions and deletions (indels), and cryptic structural variants (SVs) identified by whole genome sequencing (average coverage 28x) were validated using sample-specific custom Nimblegen capture arrays, followed by Illumina sequencing; an average coverage of 972 reads per somatic variant yielded 10,597 validated somatic variants (average 421/genome). Of these somatic mutations, 308 occurred in 286 unique genes; on average, 9.4 somatic mutations per genome had translational consequences. Several important themes emerged: 1) AML genomes contain a diverse range of recurrent mutations. We assessed the 286 mutated genes for recurrency in an additional 34 NK M1-AML cases and 9 M3-AML cases. We identified 51 recurrently mutated genes, including 37 that had not previously been described in AML; on average, each genome had 3 recurrently mutated genes (M1 = 3.2; M3 = 2.8, p = 0.32). 2) Many recurring mutations cluster in mutually exclusive pathways, suggesting pathophysiologic importance. The most commonly mutated genes were: FLT3 (36%), NPM1 (25%), DNMT3A (21%), IDH1 (18%), IDH2 (10%), TET2 (10%), ASXL1 (6%), NRAS (6%), TTN (6%), and WT1 (6%). In total, 3 genes (excluding PML-RARA) were mutated exclusively in M3 cases. 22 genes were found only in M1 cases (suggestive of alternative initiating mutations which occurred in methylation, signal transduction, and cohesin complex genes). 25 genes were mutated in both M1 and M3 genomes (suggestive of common progression mutations relevant for both subtypes). A single mutation in a cell growth/signaling gene occurred in 38 of 67 cases (FLT3, NRAS, RUNX1, KIT, CACNA1E, CADM2, CSMD1); these mutations were mutually exclusive of one another, and many of them occurred in genomes with PML-RARA, suggesting that they are progression mutations. We also identified a new leukemic pathway: mutations were observed in all four genes that encode members of the cohesin complex (STAG2, SMC1A, SMC3, RAD21), which is involved in mitotic checkpoints and chromatid separation. The cohesin mutations were mutually exclusive of each other, and collectively occur in 10% of non-M3 AML patients. 3) AML genomes also contain hundreds of benign “passenger” mutations. On average 412 somatic mutations per genome were translationally silent or occurred outside of annotated genes. Both M1 and M3 cases had similar total numbers of mutations per genome, similar mutation types (which favored C>T/G>A transitions), and a similar random distribution of variants throughout the genome (which was affected neither by coding regions nor expression levels). This is consistent with our recent observations of random “passenger” mutations in hematopoietic stem cell (HSC) clones derived from normal patients (Ley et al manuscript in preparation), and suggests that most AML-associated mutations are not pathologic, but pre-existed in the HSC at the time of initial transformation. In both studies, the total number of SNVs per genome correlated positively with the age of the patient (R2 = 0.48, p = 0.001), providing a possible explanation for the increasing incidence of AML in elderly patients. 4) NK M1 and M3 AML samples are mono- or oligo-clonal. By comparing the frequency of all somatic mutations within each sample, we could identify clusters of mutations with similar frequencies (leukemic clones) and determined that the average number of clones per genome was 1.8 (M1 = 1.5; M3 = 2.2; p = 0.04). 5) t(15;17) is resolved by a non-homologous end-joining repair pathway, since nucleotide resolution of all 12 t(15;17) breakpoints revealed inconsistent micro-homologies (0 – 7 bp). Summary: These data provide a genome-wide overview of NK and t(15;17) AML and provide important new insights into AML pathogenesis. AML genomes typically contain hundreds of random, non-genic mutations, but only a handful of recurring mutated genes that are likely to be pathogenic because they cluster in mutually exclusive pathways; specific combinations of recurring mutations, as well as rare and private mutations, shape the leukemia phenotype in an individual patient, and help to explain the clinical heterogeneity of this disease. Disclosures:Westervelt:Novartis: Speakers Bureau.

TRIB2 Gene Overexpression Is Associated with Adult Minimally Differentiated Acute Myeloid Leukemia

Abstract 4628 Background.Tribbles homolog gene 2 (TRIB2) is a pseudokinase gene belonging to a three member gene family. Tribbles gene was first identified in Drosophila melanogaster where is involved in the regulation of morphogenesis and mitosis. Likewise, mammalian homologous genes of Tribbles (TRIB1, TRIB2, TRIB3) promotes the degradation of specific transcription factors and interacts with several cell signaling mediators and modulators. Recently it has been demonstrated that TRIB2 was able to induce AML in bone marrow transplanted mouse by inducing C/EBPα proteasome-dependent degradation. Moreover, gene expression profiles data from AML patients revealed that patients carrying C/EBPα mutations clustered with those patients with up-regulated TRIB2.Aims. To test the hypothesis that TRIB2 expression was related to a differentiation degree in AML, we evaluated several AML cases based on FAB cytotype. Patients and Methods.We performed quantitative Real Time-PCR (qRT-PCR) analysis on AML patients (15 AML-M0, 14 AML-M2, 5 AML-M3, 2 M4 and 3 M5b bone marrow aspirate samples) to measure the expression level of TRIB2 in different FAB AML subtype. Four healthy bone marrows were used as reference samples. qRT-PCR was conduct using SYBR green chemistry and specific primers for TRIB2 transcript and for two housekeeping genes (B2M and IPO8) previously tested for their stability and efficiency. All AML sample were tested by conventional cytogenetic analysis and by FISH with TRIB2 specific probe. We performed, also, methylation analysis of a CpG island located in the TRIB2 promoter region by methylation sensitive restriction enzyme (MSRE) and qPCR. Briefly, DNA samples were digested with MSRE and were quantified by qPCR using specific primer pairs surrounding restriction enzymes recognition sites. Results.qRT-PCR experiments revealed that TRIB2 expression was higher (from 3 to 20 fold) in AML-M0 respect to the AML-M2 (p= 0.02), AML-M3 (p=0.01), AML-M5 (p=0.006) FAB subtypes, and references (p=0.003), respectively. Moreover, the AML-M0 cases showed a TRIB2 overexpression compared to that observed in the two AML-M4 cases (about 4 fold changes) but this difference there was not statistically significant probably because of M4 cases paucity in our series. Therefore qRT-PCR displayed a progressive decreasing TRIB2 expression along FAB subtypes. Conventional cytogenetic and FISH analysis did not show any kind of rearrangement involving TRIB2 gene. MSRE experiments revealed an higher methylation degree of a CpG island located in the TRIB2 promoter region in AML-M0 cases respect to the others FAB subtypes; moreover, methylation was significantly correlated with TRIB2 expression (r = 0.9; p = 0.0002). Conclusions.Our data showed that the TRIB2 gene overexpression was associated to AML-M0. In our cases TRIB2 dysregulation was not due to gene amplification as in other cancers. Surprisingly we observed that AML-M0 samples showed an higher methylation degree in the TRIB2 promoter region. Usually methylation cause gene silencing but our results showed that TRIB2 promoter region methylation was associated to the overexpression of the gene. Maybe methylation inhibits binding of some unknown transcriptional repressor as already seen for hTERT gene in others tumors cells. In conclusion, our data revealed that TRIB2 dysregulation seems to be linked to AML-M0 phenotype. Further studies are needed to clarify the reasons for this association. Disclosures:No relevant conflicts of interest to declare.

Prostate cancer in the elderly

AbstractBACKGROUND:The objectives of this study were to determine the frequency of metastatic (M1) prostate cancer (PC) at presentation in different age groups, to examine the association of age with PC‐specific mortality, and to calculate the relative contribution of different age groups to the pool of M1 cases and PC deaths.METHODS:Records from 464,918 patients who were diagnosed with PC from 1998 to 2007 were obtained from the Surveillance, Epidemiology, and End Results (SEER) database. The patients were categorized according to age into groups ages &lt;50 years, 50 to 54 years, 55 to 59 years, 60 to 64 years, 65 to 69 years, 70 to 74 years, 75 to 79 years, 80 to 84 years, 85 to 89 years, and ≥90 years. The cumulative incidence of death from PC was computed using the Gray method.RESULTS:The frequency of M1 PC at presentation was 3% for the group aged &lt;75 years, 5% for the group ages 75 to 79 years, 8% for the group ages 80 to 84 years, 13% for the group ages 85 to 89 years, and 17% for the group aged ≥90 years. The 5‐year cumulative incidence of death from PC was 3% to 4% for all patients with PC in any category aged &lt;75 years, 7% for patients ages 75 to 79 years, 13% for patients ages 80 to 84 years, 20% for patients ages 85 to 89 years, and 30% for patients aged ≥90 years. Although patients aged ≥75 years at PC diagnosis represented just over a quarter (26%) of all PC cases, they contributed almost half (48%) of all M1 cases and more than half (53%) of all PC deaths.CONCLUSIONS:Compared with younger patients (aged &lt;75 years), older patients were more likely to present with very advanced disease, had a greater risk of death from PC despite higher death rates from competing causes, and contributed more than half of all PC deaths. Awareness of this issue may improve future outcomes for elderly patients with PC. Cancer 2012;118: 3062–70. © 2011 American Cancer Society.

Cytoplasmic nucleophosmin (cNPM) in acute myeloid leukaemia: Relation to disease characteristics

BackgroundNucleophosmin/B23 (NPM) is a 38 kDa molecular phosphoprotein involved in ribosome assembly and transport. Findings have revealed a complex scenario of NPM functions and interactions, pointing to proliferative and growth-suppressive roles. NPM appears to be more abundant in tumour cells than in normal cells. ObjectiveThe aim of the work was to identify cytoplasmic localization of NPM using bone marrow clot biopsy and correlate it with disease and patient characteristics and the known prognostic factors, induction chemotherapy response and survival after 12 months of follow up. MethodsThe present work was undertaken on 50 cases of normal karyotype acute myeloid leukaemia classified according to modified FAB system. Bone marrow aspirates were obtained, clotted and a formalin-fixed, paraffin embedded cell block was prepared. Sections were immunostained for NPM. ResultsTwenty-six cases (52%) had cytoplasmic positivity (cNPM+) while the remaining 24 cases (48%) had nuclear restricted NPM (cNPM−). cNPM positivity was significantly variable among the different FAB subtypes being the most prevalent among M5 and M2 cases, was associated with high blast and white blood cell count at diagnosis. It was significantly correlated with CD34 negativity, CD14 positivity but not with FLT3 mutations. Clinically no relation between cNPM positivity and age, sex nor extramedullary involvement of the studied patients was proven. The cytoplasmic positivity for NPM was significantly correlated with increased survival and better outcome after cycles of chemotherapy. So it can be regarded as a good prognostic marker. FLT3 positivity affected the survival of the cNPM negative group of patients remarkably, denoting the importance of combining both their statuses to predict outcome of therapy and survival. ConclusionOur data confirm that cytoplasmic NPM1 immunoreactivity is predictive of NPM1 mutations and can be included in the routine diagnostic and prognostic workup of AML.