bluuebunny/crossref_metadata_2025 huggingface dataset:
Elevated Endothelial Progenitor Cells during Painful Sickle Cell Crisis
Authors: Nur Erfan , van Beem Rachel T. , Zwaginga Jaap Jan , Landburg Precious , van Beers Eduard J. , Duits Ashley , Schnog John-John , Lommerse Ingrid , Van der Schoot C. Ellen , de Boer Hetty C. , Brandjes Dees P.M. , Biemond Bart J.
Published: April 2020

Background: Sickle Cell Disease (SCD) is characterized by microvascular occlusion leading to an ischemia-induced pro-angiogenic response. Circulating endothelial progenitor cells (EPCs) play an important role in neovascularisation and decreased EPC numbers are associated with poor outcome in cardiovascular disease.

Aims: To determine the numbers of circulating EPCs in patients with SCD during steady state and painful crisis and to assess the association of EPC numbers with serum levels of humoral growth factors (stromal derived growth factor-1α (SDF-1α), erythropoietin (EPO), vascular endothelial growth factor (VEGF), placenta-induced growth factor (PIGF), interleuking-8 (IL-8), soluble vascular adhesion molecule-1 (sVCAM-1)).

Methods: Numbers of circulating EPCs, defined as CD45dim/CD34+ cells expressing vascular endothelial growth factor receptor-2 (VEGF-R2), and serum levels of humoral factors were measured consecutively in 66 patients in steady state, 23 patients during painful sickle cell crisis and 13 age- and race-matched healthy controls.

Results: While circulating EPC numbers were comparable between healthy controls and sickle cell patients in steady state, they were significantly increased in patients during sickle cell crisis. EPC numbers correlated significantly only to serum SDF-1α levels during painful crisis.

Conclusion: Sickle cell crisis is characterized by elevated circulating EPC numbers further supporting a pro-angiogenic state in SCD. SDF-1 may play a role of importance in EPC mobilisation during sickle cell crisis.
Identification of Multiple Oxidative Post-Translational Modifications of Plasma Albumin in Patients with Pulmonary Hypertension of Sickle Cell Anemia
Authors: Odhiambo Adam , Perlman David , Steinberg Martin , Costello Catherine E. , McComb Mark E. , Klings Elizabeth
Published: April 2020

Background: Pulmonary hypertension (PH) in sickle cell anemia (SCA) is characterized by decreased nitric oxide bioavailability; which may, in part, be related to increased oxidative stress. It is possible that via protein post-translational modifications, oxidants are able to affect both protein structure and function. We hypothesized that, in patients with SCA and PH, oxidative post-translational modifications (PTMs) occur on plasma proteins and are important in disease pathogenesis. We previously reported the identification of one PTM, a malondialdehyde adduct on peptide 146–159 of albumin and sought to do a more extensive evaluation of this protein to determine the presence of other abnormalities.

Methods: Plasma was obtained from subjects with:

SCA and PH (n=5); SCA steady-state without PH (n=4); Pulmonary Arterial Hypertension (PAH) (n=4); no evidence of cardiopulmonary disease (n=4).

Platelet-poor plasma was separated into albumin-enriched and albumin-depleted fractions. The albumin-enriched fraction was subjected to proteolytic digestion by trypsin and studied by matrix-assisted-laser desorption/ionization (MALDI) mass spectrometry (MS) and liquid chromatography (LC)-MS/MS tandem mass spectrometry. Proteomic analyses were performed on all samples and post-translational modifications characterized by MS/MS.

Results: We have characterized several additional peptides of albumin from patients with PH of SCA that bear lipid peroxidation and glycation adducts. Our comprehensive LC-MS/MS results have allowed us to identify 4-hydroxynonenal (HNE), hexose (HEX) and malonyl adduction at numerous, distinct cysteine, histidine, lysine, and threonine albumin residues.

Conclusion: Increased oxidant burden and altered redox biology is characteristic of PH in SCA. Plasma proteins, such as albumin, are a target for oxidants and changes in their structure may play a role in disease pathogenesis.
Benzocaine Induced Methemoglobinemia a Potentially Life Threatening Complication after Transesophageal Echocardiogram
Authors: Sreenivasappa Shylendra B , Yim Barbara , Catchatourian R.
Published: April 2020

Methemoglobinemia is a potentially life threatening medical condition that occurs due to increased methemoglobin production or decreased elimination. Methemoglobin is a derivative of hemoglobin in which the normal ferrous (Fe2+) iron state is oxidized to the ferric (Fe3+) state. This oxidized hemoglobin is unable to bind oxygen, hence reducing he oxygen carrying capacity of blood. The clinically presentation is characterized by cyanosis, low pulse oximeter reading and normal arterial PO2. Topical use of benzocaine a local anesthetic widely used in endoscopic procedures is one of the incriminated causes of acquired methemoglobinemia.

We present two cases of patients (pts) who developed methemoglobinemia after benzocaine spray for a transoesophageal echocardiogram. One pt was 50 year old male and other 30 year old female. Both had a methacilline resistant Staphylococcus Aureus bacteremia secondary to septic arthritis, they under went a transesophageal echocardiogram to rule out endocarditis. Topical local anesthesia used was benzocaine 20%. About 10–15 min after local anesthesia they desaturated to the mid 80’s and they developed cynosis, they were intubated. On Blood gas analysis methemoglobin. was elevated 38.4% (male pt) and 18.9% (Female pt). They were given methylene blue at the dose of 2mg/kg intravenously over 10 minutes. Cyanosis corrected with in an hour. Methemoglobin level corrected to normal after 3 hours after administration of methylene blue. There were no adverse events with administration. The Glucose 6 Phosphate Dehydrogenase enzyme level was tested and was negative.

In conclusion benzocaine induced methemoglobinemia should be considered in pts under going Transesophageal Echocardiogram when cyanosis develops that is unresponsive to oxygen. Diagnosis can be confirmed by arterial blood gas analysis. If identified early this potentially life threatening condition can be treated with methylene blue with minimal complications and good outcomes.
Prognostic Analysis of Hodgkin Lymphoma (HL) in JAPAN
Authors: Okamoto Rumiko , Itoh K. , Kinoshita T. , Watanabe T. , Yoshimura K. , Chou T. , Ogura M. , Hirano M. , Asaoku H. , Kurosawa M. , Maeda Y. , Omachi K. , Moriuchi Y. , Kasai M. , Ohnishi K. , Takayama N. , Morishima Y. , Tobinai K. , Kaba H. , Yamamoto S. , Fukuda H. , Kikuchi M. , Yoshino T. , Matsuno Y. , Hotta T. , Shimoyama M.
Published: April 2020

Background: Since the incidence of HL in Japan is approximately one-third of that in western countries, there are few studies on validation of international prognostic score (IPS) in Japanese patients (pts) with HL.

Patients and Methods: JCOG-LSG conducted 2 multicenter phase II trials for advanced HL, including ABVd (JCOG9305) and ABV followed by involved-field radiotherapy (JCOG9705). Because dacarbazine was highly emetic, and not approved for HL in Japan in those days, the dose of dacarbazine was reduced to a two-thirds (250 mg/m2) of that in original ABVD regimen in ABVd, and in ABV with increased dose of doxorubicin, dacarbazine was not utilized. Among the whole 200 enrolled pts, histopathological specimens from 181 pts were reviewed by 6 hematopathologists and consensus diagnosis of HL was made in 167 (92.3%), according to the WHO classification. Major eligibility criteria of the trials were age between 15 and 69, ECOG performance status of 0 to 3, and clinical stage of II, III or IV in JCOG9305, and IB, IIB, III or IV in JCOG 9705.

Results: 5-year survival of the 167 patients was 88.3%. Histopathological distributions of 167 pts with HL were similar with those in western countries; 2 nodular lymphocytic predominance (1.2%), 115 nodular sclerosis (68.9%), 3 lymphocyte-rich (1.8%), 34 mixed cellularity (20.1%), 7 lymphocyte depletion (4.2%), and 6 unclassified (3.6%). IPS was poorly fitted for the overall survival (OS), mainly because of too good prognosis of patients with IPS-grade 6. Seven unfavorable prognostic factors for OS identified by the univariate analysis were male sex, high β2 microglobulin, B symptoms, high LDH, high alkaline-phosphatase, clinical stage of III or IV, and histopathological subtype (mixed cellularity or lymphocyte depletion). Excluding β2 microglobulin from analysis because of only 105 available data, male sex [HR 3.30 (95%CI: 1.15–9.52, p=0.027) ] and high LDH [HR 2.41 (95%CI: 1.07–5.43, p=0.034)] were significant independent risk factors in a multivariate analysis.

Conclusions: Our study suggests that male sex and high LDH might be important prognostic factors in OS of pts with HL. Simple prognostic model for HL, including sex and LDH, was suggested.
The Significance of the Variants Erythrovirus in Patients with Cytopenias of Unknown Origin
Authors: Garcia Sheila Oliveira , Pereira Juliana , Sanabani Sabri , Neto Walter Kleine , Sabino Ester Cerdeira
Published: April 2020

Erythrovirus are now classified in three groups: genotype 1 (prototype Pvbaua); genotype 2 (Lali prototype) and genotype 3 (V9 prototype). The role of genotypes 2 and 3 in the development of disease remains uncertain. We have recently showed that those genotypes may represent 50% of the infections in Brazil. The objective of this study is to verify if these viruses are associated with cytopenia of unknown origin. We have designed a case control study in which the prevalence of erythrovirus in the bone marrow sample was compared among cases of cytopenia of unknown origin and 2 controls: bone marrow donors (C1) and, individuals with chronic leukemia (C2). For detection of the virus a nested PCR was performed in bone marrow and plasma samples. Table 1 describes the results obtained in 68 C0, 35 C1 and 67 C2 collected during the year of 2007. The prevalence of the virus in bone marrow was the same in the 3 groups. This result suggests that erythrovirus are common in bone marrow and probably not are responsible for most of cytopenias of unknown origin. A detection of this agent only in bone marrow has little clinical relevance.

Table 1. Results obtained during the year of 2007

Cases (C0) Cytopenias Control 1 (C1) Bone marrow donors Controle 2 (C2) Chronics Leukemias Bone marrow Plasma Bone marrow Plasma Bone marrow Plasma positive total positive total positive total positive total positive total positive total 6 (9%) 68 1 (1%) 68 3 (8%) 35 0 35 5 (7%) 67 0 67
Effect of the Absolute Lymphocyte Count on Induction Chemotherapy Outcomes in Adult Patients with Acute Leukemia
Authors: Daily Karen C , Hsu Jack W , Subbiah Sukanthini , Aman Shazia , Hou Wei , Wingard John R
Published: April 2020

Abstract 4853 was withdrawn on April 3, 2009 by authors' request.
Molecular Events of T-Cell Lymphoblastic Leukemia in Early Childhood
Authors: Mansur Marcela Braga , Emerenciano Mariana , Brewer Lilian , Candian Kátia Cristina da Cruz Machado , Pombo-de-Oliveira Maria S.
Published: April 2020

T-cell acute lymphoblastic leukemia (T-ALL) predominantly in children > 5 years-old, which accounts for approximately 15% of pediatric ALL in age range. Recent studies demonstrated that an accumulation of chromosomal rearrangements and other genetic anomalies is necessary for T-ALL development. However causal factors and time latency for childhood T-ALL remains enigmatic. Unlike B-cell precursor ALL that involves genetic events prenatally or in early infancy, T-ALL early childhood have controversial results regarding prenatal origin. Therefore, we conducted a molecular screening in infants with T-ALL searching for the most common genetic markers associated with this malignancy. This series of cases was selected from the incident cases included in the Brazilian Collaborative Study for IAL. The diagnosis of T-ALL was made according to morphology and EGIL classification, followed by conventional karyotyping. Sixteen cases of T-ALL IL were analyzed either by PCR-RFLP (FLT3), RT-PCR (MLL, SIL/TAL1 and HOX11L2) or PCR-DHPLC (NOTCH1), tracing the genetic alterations commonly found in these genes. Mean age at diagnosis was 14.6 months (6–24), there was a predominance of high white blood cells (WBC) count, and a poor survival in this series of cases. Cytogenetic aberrations included del(12p) (#2), t(11;14) (#3), del(6)(q23) and del(7) (q32) (#11), and 49, XY,+6,+8,+19 (#15). Details of all molecular markers are presented in Table. Then molecular markers were identified in 8 out of 16 (50%) T-ALL cases. The observed markers included MLL rearrangements (n=2), FLT3-ITD (n=1), SIL/TAL1 (n=2), HOX11L2 (n=1), and NOTCH1 (n=4). SIL/TAL1 and NOTCH1 abnormalities were found concomitantly in one case (#7) in the diagnostic sample. By the fact that the onset of T-ALL arose in a short time latency from birth to diagnosis we suggested that T-ALL might originate prenatally in these cases.

Table. Laboratorial and Molecular Features of Patients

ID Age(months) Gender WBC(×109/L) EGIL MLL FLT3 SIL/TAL1 HOX11L2 NOTCH1 Outcome #1 24 F 86.0 T-I Neg WT Neg Neg WT Dead #2 20 F 240.1 T-I Neg WT Neg Neg WT Dead #3 22 M 380.1 T-II Neg ITD Pos Neg WT Dead #4 24 M 440.1 T-IV Neg WT Neg Neg WT Dead #5 20 F 330.0 T-IV Pos1 WT Neg Neg WT Alive #6 12 M 80.1 T-IV Neg WT Neg Neg WT Dead #7 13 F 71.3 T-I Neg WT Pos Neg Mut Lost #8 9 M 56.2 T-IV Neg WT Neg Neg Mut Alive #9 14 M 57.1 T-IV Neg WT Pos Neg WT Lost #10 7 M 110.0 T-IV Pos2 WT Neg Neg WT Alive #11 17 M 180.1 T-I Neg WT Neg Neg WT Alive #12 20 F 78.6 T-I Neg WT Neg Neg WT Dead #13 9 M 71.0 T-IV Neg WT Neg Neg WT Dead #14 9 F 50.0 NA Neg WT Neg Neg Mut Dead #15 8 M 154.0 T-IV Neg WT Neg Neg Mut Alive #16 6 F 131.6 T-IV Neg WT Neg Pos WT Alive

Abbreviations: F, female; M, male; WBC, white blood cells count; NA, not available; Neg, negative; Pos, positive; WT, wild-type; ITD, internal tandem duplication; Mut, mutated; 1MLL/AF4; 2MLL/ENL.
Long-Term Follow up of Chinese Children with Acute Lymphoblastic Leukemia Based on Minimal Residual Disease Detection
Authors: Tang Yongmin , Xu Xiaojun , Shen Hongqiang , Song Hua , Shi Shuwen , Yang Shilong , Wei Jian , Xu Weiqun
Published: April 2020

Introduction and objective: Recent studies from resource-rich countries have showed that minimal residual diseases (MRD) were the most important independent prognostic factor for children with acute lymphoblastic leukemia (ALL). However, experience from Chinese children with this disease has not been reported in the English literature. In this study, we reported our experience in small series of children with ALL based on multi-parameter flow cytometric detection of MRD to study the relationship between MRD levels and the long term survivals in Chinese children with ALL.

Methods: Using multi-parameter flow cytometry and at least one patient specific leukemia associated immunophenotype (LAIP) to determine the MRD levels in 71 Chinese children with ALL expressing leukemia associated immunophenotype (LAIP) from January 2002 to December 2003. Bone marrow aspirates were collected on day 15 and the 1st, 3rd, 6th, 12th, 18th, 24th, 30th month after chemotherapy. Leukemia associated Immunophenotype for each patient was identified according to the patients’ initial immunophenotypes determined by multi-parameter flow cytometry using a panel of more than 30 antibodies.

Results: The one, two and three year disease free survival (DFS) for patients with a MRD level less than 10−2 on day 15 were (88.3±6.4) %, (84.3±7.2) % and (76.3±8.5) %, respectively, whereas those for patients with MRD levels ≥10−2 on day 15 were 100%, 0% and 0%, respectively (log rank test, P=0.004). The 5-year DFS for patients with MRD levels <10−2 and ≥10−2 on day 15 were (83.8±6.1) % and 0%, respectively (P<0.01). The 5-year DFS were significantly different between patients with MRD levels <10−4 and those ≥10−4 on the 6th or 12th month. Out of 71 patients, the 5-year DFS for patients with MRD levels <10−3 was 100%, however, the 1-year, 3-year and 5-year DFS for patients with MRD levels ≥10−3 were (87.3±5.3) %, (71.4±7.3) % and (68.0±7.7) %, respectively (log rank test, P=0.004). And the 1-year, 3-year and 5-year relapse rates for the latter group were 10.5%, 13.5% and 15.8%, which showed a tendency of difference in terms of relapse rate when compared to patients with MRD levels <10−3 (5-year relapse rate: 15.8% vs. 0%, P=0.08).

Conclusions: MRD detected by multi-parameter flow cytometry is also an independent prognostic factor in Chinese children with ALL.
Short Time Latency in Infant Leukemia with ETV6/RUNX1 fusion Gene
Authors: Emerenciano Mariana , Bungaro Silvia , Cazzaniga Giovanni , Dorea Maria Dolores Fonseca , Coser Virginia Maria , Magalhaes Isis Quezado , Biondi Andrea , Pombo-de-Oliveira Maria S.
Published: April 2020

Acute lymphoblastic leukemia (ALL) in infants is characterized by a high frequency of MLL gene rearrangements. By contrast, the ETV6/RUNX1 (TEL/AML1) fusion gene is usually detected in children older than 2 years. In a series of Brazilian infant leukemia cases collected in an epidemiological study, a screening was performed to identify fusion genes found in ALL, we have identified four cases harboring ETV6/RUNX1 in blast cells of infants with 2, 3, 5 and 7 months-old, and common-ALL. To underlie additional genomic hits required to accelerate the emergence of a frank leukemia in these c-ALL cases, we applied the SNP-based genomic copy number analysis, comparing t(12;21) infants with other children with c-ALL in older age-range groups. The presence of ETV6/RUNX1 fusion gene was demonstrated by reverse transcription polymerase chain reaction (RT-PCR) and confirmed by fluorescence in situ hybridization (FISH). All cases were negative for MLL rearrangements, as assessed both by RT-PCR and FISH analyses. As control group (non-leukemic children), we selected 5 samples from children younger than 2 years-old already included in a previous epidemiological study. Infant cases described here present high WBC count (on average 131 × 109/L), and adverse prognosis: two of them relapsed, and one patient only (relapsed) is still alive four years after diagnosis. Recurrent deletions, including 9p21.3 (CDKN2A/2B and MTAP), 11p13 (CD44), 12p13.2 (ETV6) and regions affected by copy number neutral loss of heterozigosity (LOH) were observed. The frequency of genomic deletions and amplifications detected by 100K SNP arrays varied significantly between age-related c-ALL subgroups. The infant leukemia group showed the lower number of chromosome imbalances, whereas the mean number of deletions was similar in the <2 years and 2–10 years age groups, consistent with the observation that the number of gains was largely increasing in older children. It has been postulated that infant leukemia might be caused by transplacental exposures during pregnancy. Then, with these unusual results we have reviewed the data of their mothers’ interviews. Mothers of infants were effectively exposed to a non-steroidal anti-inflammatory drug, and other chemical substances. It may be hypothesize that the pre-leukemic phase in these age-related subgroups of ETV6/RUNX1 positive patients may be evolved with different latencies due to the occurrence of a combination of genetic and environmental factors.
Identification of New Markers Discriminating Between Myeloid and Lymphoid Acute Leukemia
Authors: Haouas Houda , Haouas Samira , Uzan Georges , Hafsia Aicha
Published: April 2020

The heterogeneity of acute myeloid leukemia (AML) with respect to biology and clinical course resides in the fact that patients belonging to the same group show marked differences in their response to chemotherapy, which would necessitate a refinement of AML classification. In order to contribute to define molecular markers for AML we realized microarray assays on two M5 AMLs and selected four differentially expressed genes to validate their expression by RQ-PCR. We have shown that two down-regulated genes in AML, Guanine nucleotide binding protein (G protein) gamma 11 (GNG11) and Amphiregulin are also down-regulated in B-ALL and T-ALL patients. We have found a gene, Ceruloplasmin, which is up-regulated in AML but not in B-ALL and T-ALL. The level of expression of these genes varies from one patient to another. Since the group of patients studied is limited, further studies must carry on with a larger series of patients to be able to make subdivision according to the expression of GNG11, Amphiregulin and Ceruloplasmin. Our study is the first to analyze these genes in AML, B-ALL, T-ALL and CL patients by quantitative real time PCR. This rapid and sensitive method could be used to screen these genes in different types of leukemia.
Acute Lymphoblastic Leukemia with Mature Appearing Lymphocytes: First Chinese Case Report and Literature Review
Authors: Zhang QiGuo , Ouyang Jian , Li Jianyong
Published: April 2020

Objective: To increase the knowledge and understanding of acute lymphoblastic leukemia with maturation(ALLm).

Method: One ALLm case with clinical manifestation, bone marrow morphology, immunophenotype and cytogenetic results were presented and related literatures were reviewed.

Result: The patient was a fifty-five year old women, the haematological feature was Pancytopenia. There were 12% lymphoblasts and 82.5% mature appearing lymphocytes in the bone marrow smear. The mature appearing leukemic cells could not be separated clearly by gating. However, the immunophenotypes of mature-appearing leukemic cells(Low FSC and Low CD45) and lymphoblasts were the same. Both results were CD33+CD34+CD19+CD22+HLA−DR+CD5−CD7−CD10−CD13−CD14−CD15−CD20−CD25−CD45−CD71−CD11b−CD103−CD117−. Bone marrow biopsy showed hypercellularity with diffuse infiltration of lymphocytes, The results were CD34++,TdT++,Pax-5+++,CD43++,CD3+(scatter),CD5+(scatter),CD20−,CD10+,CyclinD1−, lymphoblasts Ki67+, mature-appearing leukemic cells ki-67-. FISH analysis of the bone marrow revealed about 1% cells with a signal pattern suggesting loss of one copy of chromosome 8. After VDP, MA, AAG and HAG regimens Complete remission was not achieved.

Conclusion: ALLm is a special morphological variant of acute lymphoblastic leukemia, most mature appearing cells are in resting G0 phase and this could be the reason why ALLm has a poor response to chemotherapy.
Long-Term Increases in Lymphocytes and Platelets in Human T-Lymphotropic Virus Type II Infection
Authors: Bartman Melissa T , Kaidarova Zhanna , Hirschkorn Dale , Sacher Ronald A. , Fridey Joy , Garratty George , Gibble Joan , Smith James W , Newman Bruce , Yeo Anthony E , Murphy Edward L
Published: April 2020

Human T-lymphotropic viruses types I and II (HTLV-I and HTLV-II) cause chronic infections of T-lymphocytes, leading to adult T-cell lymphoma (HTLV-I) and myelopathy (both types) in a minority of infected humans. However, their long-term effects on blood counts and hematopoiesis are not fully understood. We followed 151 HTLV-I and 387 HTLV-II seropositive former blood donors, and 799 HTLV seronegative donors from five US blood centers prospectively for a median of 14.0 years. Complete blood counts were performed every 2 years on fresh anticoagulated blood at licensed clinical laboratories near each center. Multivariable repeated measures analyses were conducted to evaluate the independent effect of HTLV infection and potential confounders on 9 hematologic measurements. HTLV-II subjects had significant (p<0.05) increases in their adjusted lymphocyte counts (+126 cells/mm3; approx +7%), hemoglobin (+0.2 gm/dL) and mean corpuscular volume (MCV; 1.0 fL) compared to seronegative subjects. Both HTLV-I and -II subjects had higher adjusted platelet counts (+16,544 and +21,657 cells/mm3; p<0.05) than seronegatives. Among all subjects, time led to decreases in platelet count (−6,750 to −6,600 cells) and lymphocyte counts (−67 to −66 cells), and to increases in MCV (+0.4 fL) and monocytes (+19 to +20 cells per uL; all per two-year interval). Women had lower hemoglobin but higher platelet and white blood cell (WBC) counts than men. Blacks had lower hemoglobin, MCV and neutrophil counts, but higher platelet and lymphocyte counts than whites. Heavy drinking was associated with higher MCV but lower WBC counts, whereas cigarette smoking was associated with higher WBC counts of all lineages. These results suggest that HTLV-I and -II infection can produce significant, independent and long term changes in lymphocytes, platelets and red blood cells. The finding of increased lymphocytes in HTLV-II infection is novel and may be related to viral transactivation or immune response. HTLV-I and –II associations with higher platelet counts suggest viral effects on hematopoietic growth factors or cytokines.
Relationship Between Galectin-10 Expression and Severity of Celiac Disease Abolished in the Presence of Gammadelta-Positive T-Cell Clonal Expansion
Authors: Simula Maria Paola , De Re Valli , De Zorzi Mariangela , Canzonieri Vincenzo , Spina Michele , Cannizzaro Renato
Published: April 2020

Celiac disease (CD) patients are prone to develop T-cell lymphoma throughout a progressive accumulation of aberrant, clonal gd+ T-cells. It is supposed that high production of pro-inflammatory cytokines, and chronic antigenic stimulation, due to gluten ingestion, plays a key role in inducing inflammation, resistance to apoptosis and the emergence of these T-clones. Regulatory T-cells maintain immunological self-tolerance by active suppression of auto aggressive T-cells. Among them, Tr1 subset plays a role in the suppression of naïve and memory T-cells. The role of Tr1 in human diseases is not well understood, even because there are no specific markers able to identify these cells, however recently, Galectin-10 has been proposed as a marker for functional Tr1. To better understand pathogenetic mechanisms associated with CD and clonal T-cell proliferations we investigated galectin-10 expression from gut epithelium by 2D-DIGE approaches. Patients were selected and grouped for histological inflammatory degree and for gd+T pattern defined by g-TCR genescan analysis. Groups consisted of 7 individuals with Marsh-0 (4/7 oligoclonal), 3 with a Marsh-1 or -2 (3/3 polyclonal) and 5 with Marsh-3 (2/5 clonal gd+T). Control consisted of 4 individuals with excluded CD. We found, a parallel increase in galectin-10 levels and Marsh index in individuals with polyclonal gd+T cells (p=0.0092), while reduced levels were evidenced from patients with clonal gd+T cells and Marsh-3 (p=0.017). We assume that galectin-10 up-expression is induce to an attempt to extinguish inflammation. If these clones are those more susceptible to malignant progression reserve further exploration.
CD3-Zeta Gene Expression in Workers Benzene-Exposed and Benzene- Poisoned Workers
Authors: Li Bo , Li Yangqiu , Chen Shaohua , Yang Lijian , Yu Wei , Liu Weiwei , Chen Jiayu
Published: April 2020

Benzene is an industrial chemical and component of cigarette smoke, gasoline, and automobile emissions. Benzene’s toxic effects on the blood and bone marrow can induce aplastic anemia and leukemia. Benzene is known to be highly toxic to a variety of cell types including lymphocytes. Our previous study showed that skewed distribution and clonal expansion of TCR Vα subfamily T cells had been found in benzene-exposed workers, indicating that the disorder of T cell immune function might relate to benzene-exposed. The TCR expressed on the surface of T cells is associated with an invariant structure-CD3 and composed the TCR/CD3 complex. The CD3ζ chain plays an important role in the complex which involved in signal transduction. Little is known about the feature of CD3 ζ chain expression in benzene-exposed workers. To further identify the expression level of CD3 ζ gene in benzene-exposed workers, real-time PCR with SYBR Green±technique was used for detecting CD3 ζ gene expression level in peripheral blood mononuclear cells from 29 benzene-exposed workers, 42 benzene-poisoned workers and 20 normal individuals. β2- microglobulin gene (β2M) was used as an endogenous reference. Relative changes in CD3 ζ gene expression level was used by the 2−ΔCt×100% method (ΔCt=Ct(ζ) −Ct(β2M) ). CD3ζ gene could be detected in all of normal individuals, however, only 21 out of 29 benzeneexposed workers could be detected the CD3ζ gene with a mean expression level of 15.0 ±24.9, and in 33 of all 45 benzene-poisoned workers with mean value of 19.8 ±29.7. The detectable CD3 ζ gene expression level in both benzene-exposed and benzene-poisoned groups increased significantly compared with that in normal individuals (3.00±2.11, P< 0.05). This is, to our knowledge, the first description of the effect of benzene-exposed on the expression of the CD3 ζ gene. The abnormality expression of CD3 ζ gene might lead to immune dysfunction in benzene-exposed and benzene-poisoned workers. In addition, CD3ζ gene could not be detected in a part of samples, whether the absence of CD3ζ gene might related to dysfunction of T cells in workers with benzene-exposed and benzenepoisoned, it remains an open question.
A Prospective, Observational Study on the Incidence of Chemotherapy-Induced Neutropenia in Lymphoma Patients
Authors: Salar Antonio , López Andrés , Torres Juan Pío , López María Dolores , Prieto Elena , Caballero Dolores , Giraldo Pilar , Batlle Montserrat , Benedit Patricia , Garrido Teresa
Published: April 2020

Background. Neutropenia is a common toxicity in patients (pts) with lymphoma (lymph) who receive myeloablative chemotherapy (CT). It frequently leads to CT delays and dose reductions, potentially compromising the clinical outcome. Granulocyte colonystimulating factors (G-CSFs) represented a major development in the prevention of this disorder. Current European and US guidelines (2006) recommend primary prophylaxis with G-CSF for patients at overall ≥20% risk of febrile neutropenia (FN) due to CT and patient-related factors.

Methods. A multicentre, prospective, observational study, in adult pts with lymph initiating a new CT regimen with at least 4 planned cycles, assessing the incidence of grade 3–4 neutropenia (G3–4N) [defined as absolute neutrophil count <1.0 × 109/L] over the first four cycles of CT regimens with high or intermediate FN risk (FN risk ≥20% or 10–20%, respectively).

Results. This interim analysis contains data from 270 consecutive lymph pts (300 pts per protocol) from 31 Spanish centres from November 2005 to November 2007. Pts were 53.3% male, median age 57.5 years (range: 19–85), 87.0% ECOG 0-1 and 60.9% III–IV stage (43.6% IV stage). 71.8% of lymph pts were treated with CHOP-based CT (83.0% R-CHOP). G-CSF was used in 83.9% of pts (76.8% primary prophylaxis (PP), 23.2% secondary prophylaxis (SP)). The G-CSF received was 49.1% filgrastim and 50.9% pegfilgrastim. Global incidence of G3-4N over the first four cycles was 39.9%. The G3-4N incidence was 39.6% in pts treated with pegfilgrastim while it was 52.3% in pts treated with filgrastim. Pts treated with PP had an FN incidence of 15.4% while the incidence was 22.0% in those receiving SP. Full dose on schedule (FDOS) [defined as ≤ 15% dose reduction and ≤ 3 days dose delay] was achieved in 65.1% of pts treated as PP and 60.8% of pts treated as SP.

Conclusion. This study of clinical practice suggests that current guideline recommendations on the use of G-CSF PP with CT are becoming widely adopted in Spain. In patients receiving CT with intermediate/high FN risk, G-CSF PP and pegfilgrastim prophylaxis seemed to reduce neutropenia events compared with SP and Filgrastim. PP also improved the delivery of CT at full dose on schedule.
Intrapleural Rituximab (R) Is Effective in Refractory Pleural Effusion Due to Non- Hodgkin’s Lymphoma
Authors: Rivas-Vera Silvia , Cadena-Eumaña Carlos , Chalapud-Revelo Juan , Aguilar-Navarro Adriana , Sobrevilla-Calvo Pedro , Lievano-Torres Cristina
Published: April 2020

We report the case of a 65 years old woman with a NHL, CD20 +, BCL-2 negative, Grade III, CS: IVBX, FLIPI High Risk, diagnosed on September 2005. The patient was treated with several chemotherapy regimens (CHOP, chlorambucil/prednisone, cyclophosphamide/prednisone), while on treatment a large pleural effusion was found on a chest X-ray. On pleural fluid cytology we observed numerous abnormal lymphocytes. We did repeated thoracocentesis without success, finally a pleural tube was inserted and intrapleural bleomycin instilled, again with no success (Table 1). Six weeks later, after signing informed consent, we administered 50 mg of intrapleural R (50 mg in 50 cc of SS 0.9%) without immediate or delayed adverse events. On day +4 a new chest X-ray showed marked improvement. We planned to repeat the dose every week × 4, unfortunately the patient could not afford it. After seven months the patient is asymptomatic and without recurrence of the pleural effusion.

Table 1

Date (2007) Fluid drained (ml) Action September 12 Dyspnea September 21 1,500 October 4 1,500 October 6 570 October 8 400 October 12 20 Bleomycin 10 U November 6 Dyspnea/1,300 November 9 40 November 16 400 November 17 200 November 20 100 Rituximab 50 mg

Discussion Although the pharmacology of IV R is welll known, there is scarce information about dosage, penetration into the tissues and efficacy of intracavitary R. It is likely that the intrapleural instillation of R was effective in the control of this complication in a definitive way, without adverse effects, Our case supports the results reported by Schmidt et al (Table 2). The mechanism of action at the local site is not known, but it could be the same as described with the systemic use. Pleural effusion secondary to refractory lymphoma is a rare complication, we propose the integration of an international registry of this cases to study the pharmacokinetics, pharmacodynamics, efficacy and safety of intrapleural rituximab.

Table 2

Features Our patient Schmidt Patient Age age 65 years 57 years Diagnosis Follicular NHL, Grado II Lymphoplasmacytic NHL No. Previous Therapy 3 1 Effusion therapy Thoracocentesis and drainage by pleurostomy Thoracocentesis Rituximab Dose 50mg, single dose 50mg day 1 200mg day 2 400mg day 3 Time to Response 4 days 3 days
Increased ABCG2 Expression Could Be Responsible for Resistance to Imatinib Mesylate in Patients with Chronic Myeloid Leukemia Who Do Not Have Mutations in BCR-ABL Kinase Domain
Authors: Balasubramanian Poonkuzhali , Markose Preetha , Chendamarai Ezhilarasi , Mathews Vikram , Srivastava Vivi , Velayudhan Shaji R , Lakshmi Kavitha M , Viswabandhya Auro , George Biju , Srivastava Alok , Chandy Mammen
Published: April 2020

Mutation in the Bcr-Abl kinase domain is one of the most common mechanism of resistance to imatinib mesylate (IM), seen in 30– 90% of patients with chronic myeloid leukemia (CML). A total of 57 patients with CML, 41 in chronic phase, 10 in accelerated phase and 6 in blast crisis, were analyzed for bcr-abl kinase domain (KD) mutations using reverse transcriptase polymerase chain reaction (RT-PCR) amplification of the bcr-abl KD followed by direct sequencing. Twelve different mutations including three novel mutations were identified in the bcr-abl KD in 18 patients. Resistance to IM was considered if the patient had rising white blood cell counts or did not achieve hematological remission at 6 months; or had rising bcr-abl levels (as determined by fluorescence in situ hybridization or real-time quantitative PCR) after the start of IM treatment. In order to evaluate other potential mechanisms of resistance in patients without mutations in the bcr-abl kinase domain, we tested the expression levels of transporter genes known to be involved in IM influx and efflux. mRNA expression levels of efflux transporters, ABCG2 and ABCB1, and influx transporter, hOCT1 were measured by real time quantitative PCR using ABL to normalize the expression levels of the same. Serially diluted cDNA made from HepG2 RNA was used to make the standard curve and amplification efficiencies of the target and the house keeping genes were similar. Each sample was analyzed in duplicate and the experiment was repeated twice. In the patients without mutations in the bcr-abl KD, significantly higher ABCG2 mRNA levels were observed compared to patients with mutations (Table). Transcript levels of ABCB1, hOCT1 or bcr-abl were not significantly different between the two groups. This study suggests that over expression of ABCG2 may be one of the mechanisms of resistance to imatinib in patients without mutations in bcr-abl. Future studies should not only compare the expression of these transporters at diagnosis (before the start of IM treatment) but also at the time of clinical resistance. This will help understand the influence of expression levels of these transporters in achieving haematological or molecular response to increased IM doses.

Bcr-abl mutation positive (n=18) Bcr-abl mutation negative (n=39) p value ABCG2: median (range) 0.0126 (0.0004–0.45) 0.051 (0.0041–0.51) 0.034 ABCB1: median (range) 0.1842 (0.00349–0.51) 0.1819 (0.0463–0.68) NS hOCT1: median (range) 488 (3.4–3394) 486 (62–3059) NS BCR-ABL: median (range) 53.96 (5.73–179.1) 51.3 (2.36–129) NS
The Effect of Phenylhexyl Isothiocyanate on Drug Resistance of K562/A02 Cell Line
Authors: Chen Bao-An , Yuan Peng , Cheng Jian , Gao Feng , Ding Jia-Hua , Gao Chong , Sun Yun-Yu , Wang Jun , Zhao Gang , Song Hui-hui , Bao Wen , Shao Ze-ye , Xia Guo-hua , Lu Quan-yi , Ma Xu-dong , Liu De-long , Wang Jue-qiong , Pei Xiao-ping , Chen Ning-na
Published: April 2020

Objective: To investigate the effect of 6-phenylhexyl isothiocyanate (PHI) on drug resistance and sensitivity on K562/A02 cell line to ADM and elucidate the probable mechanisms.

Methods: We measured growth inhibition of ADM on K562/A02 cell line by MTT assay. Apoptosis rate of K562/A02 cell line, the change of intracellular ADM and MRP1 protein level were detected by Flow Cytometry. Intracellular deoxidized GSH by spectrometric enzyme assay and MRP1 mRNA by RT-PCR semiquantitative assay were observed anteroposterior using PHI.

Results: PHI can enhance the sensitivity of K562/A02 cell line to ADM, survival rate of K562/A02 cell line decreased with the increasing concentration of PHI and ADM. Apoptosis rate increased with treating by combination of two above drugs, and multiple of drug resistance had statistical significance (P<0.05) when the concentration of PHI was more than 20μmol/l. Intracellular GSH of K562/A02 cell line reduced 5% when 1μg/ml ADM was single used, and when more than 10μmol/l PHI was used it increased slightly at first then decreased. When more than 20μmol/l PHI and 1μg/ml ADM were used combination, intracellular GSH of K562/A02 cell line decreased progressively with increasing the concentration of PHI. Protein and gene level of MRP1 have no statistical significance (P>0.05) no matter after or before PHI was used on different concentration.

Conlusion: The depletion effect of PHI on the intracellular GSH can not only partly enhance the reverse effect of ADM, but also enhance the sensitivity of K562/A02 cell line to ADM. Such depletion may diminish side effect and treatment dosage of ADM. It provides a new view to the therapy of leukaemia.
Treatment of Lower Risk MDS with Del 5q by Lenalidomide, with or without G-CSF: Current Results of the French Patient Named Program (ATU)
Authors: Lebras Fabien , Sebert Marie , Eclache Virginie , Lamy Thierry , Delaunay Jacques , Visanica Sorin , Dreyfus Francois , Banos Anne , Blanc Michel , Besson Caroline , Thomas Xavier , Rodon Philippe , Vey Norbert , Gardembas Martine , Bauduer Frederic , Ramee Jean-Francois , Delmer Alain , Rea Delphine , Turlure Pascal , Beyne-Rauzy Odile , Guerci Agnes , Ceccaldi Joel , Dugay J. , Fenaux Pierre , Ades Lionel
Published: April 2020

Background: LEN is very effective for the anemia of lower risk MDS with del 5q. A patient named program was launched by the French health agency (AFSSAPS) to use LEN in low and int 1 risk MDS with del 5q and transfusion dependence (> 2 RBC units/2 months).

Methods: Patients received 10 mg of LEN/day, 3 weeks on, 1 week off. Hematological response (IWG 2006) was assessed after 8, 16 and 32 weeks of treatment. G-CSF use was recommended in case of grade 3 neutropenia, in order to avoid dose reduction, especially during the first 16 weeks of treatment.

Results: 71 pts from 35 centers were evaluable for response as of Aug 1st, 2008. Median age was 75 [range 43–93], 68% females, median interval from diagnosis to LEN treatment 28.6 months (range 1–235). At inclusion, 32 pts had del 5q syndrome, 10 RA, 13 RARS, 2 RCMD-RS and 14 RAEB-1. IPSS was low in 43% and int-1 in 57%. Del 5q was isolated, with 1 additional and > 1 additional abn in 76%, 15% and 9% pts respectively. 23 pts were untreated, while 48 had previously received EPO. Median transfusion requirement was 4 units/2 months. 50/71 (69%) pts achieved erythroid response (IWG 2006), of whom 48 (67%) achieved transfusion independence (TI). Median time to TI was 12 weeks (range 8 – 32). TI was achieved in 76%, 62%, 68%, 70% and 50% pts with IPSS low, IPSS int 1, isolated del 5q, del 5q+1abn, del5q + >1abn, resp, and 70% and 65% of EPO pretreated and EPO naive pts (p=NS). Grade III-IV neutropenia and thrombocytopenia were seen in 86% and 26% pts resp, leading to transient discontinuation followed by reduction of LEN dosing in 55%pts. Of the 19 pts who received GCSF, 3 (16%) required transient LEN discontinuation due to neutropenia compared to 15 (28%) of the 52 who did not receive GCSF (p=0.3). Apart from neutropenia and thrombocytopenia, grade III–IV side effects were Lyell’s syndrome (n=1), deep venous thrombosis (n=3) Quincke’s oedema (n=1), No patient died from sepsis due to neutropenia or bleeding due to thrombocytopenia. With a still short median follow up on treatment (28 weeks), no relapse had occurred. 1 pt (RAEB 1 with 7% of blasts, isolated del 5q and IPSS INT-1 at inclusion who had achieved TI) progressed to AML at week 48.

Conclusion: Our results confirm on a very multicenter basis the efficacy of LEN on the anemia of lower risk MDS with del 5q,. The use of G-CSF during the early treatment phase may somewhat reduce the proportion of early LEN discontinuation due to neutropenia.
Development of a Multiplex PCR Assay for the Detection of Genomic Copy Number Changes in Myelodysplastic Syndromes
Authors: Stamatoullas Aspasia , Waultier Agathe , Jardin Fabrice , Callat Marie Paule , Parmentier Francoise , Penther Dominique , Tilly Herve , Bastard Christian
Published: April 2020

Background: Myelodysplastic syndrome (MDS) is a heterogeneous group of disorders characterized by ineffective hematopoiesis. Clonal cytogenetic aberrations are found in about 50% of primary MDS and 80% of secondary MDS. The International Prognostic Scoring System (IPSS) introduces cytogenetic abnormalities as an independent prognostic factor for both survival and progression to AML. However, conventional cytogenetics or FISH substitutes are not available for all myelodysplastic patients. In order to overcome this problem, we developed a multiplex PCR assay (quantitative multiplex PCR of short fluorescent fragments or QMPSF) measuring the copy number of genes located in regions involved in myelodysplastic syndromes on chromosome 7 and 8 and on the 5q, 12p, 17p and 20q regions.

Material and method: In order to select target region, high resolution CGH (Agilent, 44K) was performed in 12 patients (5 complex karyotypes, 2 isolated monosomy 7, 2 isolated trisomy 8, 1 isolated 17p deletion, 1 isolated 20q deletion and 1 normal karyotype) for which DNA extracted from CD34+ sorted cells was available. Twenty three primer pairs were selected on 6 target chromosomes (5, 7, 8, 12, 17 and 20) in regions either commonly deleted or involved in biallelic deletions. A QMPSF assay was developed and applied on the first 12 patients. This assay was then applied on a series of 60 patients for which DNA extracted from either CD34 + sorted cells or total bone marrow (BM) cells or peripheral mononuclear cells (PB) was available.

Results: As previously reported by several groups, the correlation between cytogenetics and CGH was very good although high resolution CGH was able to detect small cytogenetically not detected defects in most patients. For the 23 QMPSF targeted genes, the correlation between CGH log ratio and QMPSF ratio was excellent. Using QMPSF, genomic copy number changes were detected in 21/27 CD34+ samples, 16/20 BM samples and 15/20 PB samples. However, when different samples were available for the same patient (CD34, BM and/or PB) the correlation between samples was not perfect and mainly depends on the MDS subtype, the % of blasts and the presence of cytogenetics abnormalities.

Conclusion: These results demonstrate that a single multiplex PCR assay targeting 23 genomic regions located on the 6 chromosomes most frequently involved in MDS can be helpful for the prognostic evaluation of MDS patients for whom cytogenetic data are not available, and that DNA extracted from PB or total BM cells can be informative in a substantial number of cases.
Clonal Heterogeneity in the 5q- Syndrome: NPMc+ and p53 Expressing Progenitors Are Insensitive to Lenalidomide Treatment and Expand at Disease Progression
Authors: Jädersten Martin , Saft Leonie , Pellagatti Andrea , Göhring Gudrun , Fernández-Santamaria Cristina , Wainscoat James S , Boultwood Jacqueline , Porwit Anna , Schlegelberger Brigitte , Hellström-Lindberg Eva
Published: April 2020

Clonal heterogeneity has not been described in patients with myelodysplastic syndrome (MDS) of the 5q- syndrome subtype, for which lenalidomide has emerged as a highly potent treatment. Interestingly, transformation to acute myeloid leukemia (AML) occurs more frequently in patients without a cytogenetic response to lenalidomide. We describe two patients with classical 5q- syndrome with complete erythroid and partial cytogenetic response to lenalidomide, who evolved to high-risk MDS and AML, respectively, with complex karyotypes including del(5q). Immunohistochemistry of pre-treatment marrow biopsies revealed small fractions of progenitors with aberrant cytoplasmic nucleophosmin (NPMc+) expression and in one patient also a TP53 mutation. Both lesions may induce a state of genomic instability, and represent novel findings in 5q- syndrome. These subclones remained stable during response, but expanded at transformation, suggesting that pre-existing cells with more malignant genotype may be insensitive to lenalidomide and responsible for disease progression. We are currently investigating a larger patient material for the presence of subclones with molecular lesions in order to assess the potential impact on the risk of leukemic evolution.