bluuebunny/crossref_metadata_2025_split huggingface dataset:
Validation of JAK2 and New Clinical Criteria for the Diagnosis of Polycythemia Vera (PV).
Authors: Silver Richard T. , Jones Amy V. , Feldman Eric J. , Roboz Gail J. , Ritchie Ellen K. , Cross Nicholas C.P. 
Published: April 2020

Abstract
Although the pervasive presence of JAK2 in patients (pts) with PV has been noted in at least 7 published studies, its frequency has ranged from 66 to 97%. Although part of the reason for this wide range is false negative results due to the low sensitivity of sequence analysis, varying criteria used for the clinical diagnosis of PV are also likely to be an important factor. In more than 300 studies, the criteria of the Polycythemia Vera Study Group has been used, although many authors, including ourselves, have commented on their inadequacy. Alternative classifications have been the subject of debate mostly centered about using the rbc-Cr51 mass. Our criteria included the use of CR51 labeled red cells to establish first, an increase in red blood cell volume and second, I125 to establish an increase in plasma volume. Exception was made for men with a HCT ≥60%, and women ≥56%. If hypervolemic, the major causes of secondary polycythemia were excluded: congenital polycythemias and those secondary to impaired oxygenation, or benign or malignant tumors. This implied a serum erythropoietin level of ≤5u/ml. The aforementioned and 3 of the following 5 were required: splenomegaly, WBC, ≥12,000/ul, platelet count ≥600,000/ul, abnormal marrow histology. We used them to clinically diagnose 61 patients subsequently undergoing therapeutic trials whose blood was analyzed for the JAK2 V617F mutation by allele specific PCR from 679 patients with myeloproliferative diseases, including normal controls. Of 61 patients, with the phenotypic characteristics of p. vera, 59 (97%) were positive for the JAK2 mutation. The 2 remaining patients, although phenotypically characteristic of PV, were negative on repeat examinations. One pt was newly diagnosed and untreated and the other had a 6 year remission after rIFNα followed by a 1 year remission on imatinib. We conclude that determination of JAK2 is consistent with PV in virtually all cases clinically diagnosed appropriately. Our data, to be presented, indicate that the combination of increased red cell mass, demonstration of JAK2 and increased platelet count establish the diagnosis of PV in virtually all cases.
Coexistence of Mutated and Unmutated IgVH Forms in Patients with Chronic Lymphocytic Leukemia.
Authors: Sheikholeslami M. , Ma W. , Uyeji J. , Tornyos K. , Cone L. , Mamus S. , Keating M. , Albitar M. 
Published: April 2020

Abstract
The mutation status of the immunoglobulin heavy chain variable region (IgVH) gene is an independent prognostic indicator in patients with chronic lymphocytic leukemia (CLL). Unmutated IgVH is associated with rapid disease progression and shorter survival. The assay for determining IgVH mutation status depends on specific amplification of the mRNA of the expressed clonal IgVH. The presence of more than 1 clone in patients with CLL has been speculated and can be documented when the 2 clones express different light chains. Determining the IgVH gene family that is expressed in subclones also allows the confirmation of the presence of two clones in some cases. We describe 3 patients with two clones of CLL discovered in the process of determining the IgVH mutation status. PCR amplification of the expressed IgVH mRNA yielded 2 distinct bands. Sequencing each band separately revealed 2 different clones in each patient. Interestingly, each clone had a different mutation profile. In 2 of the 3 patients, the 2 clones were from completely different IgVH gene families (VH1-2 with VH2-5 and VH4-34 with VH5-51); both of these patients showed kappa light chain restriction. In the third patient the expressed IgVH gene family was VH3-21 in both clones, but 1 was mutated (6.8%) and the second was unmutated. Two of the 3 patients had evidence of aggressive disease with hepatosplenomegaly and lymphadenopathy; 1 of these patients expressed the VH3-21 gene family, which is known to be associated with aggressive disease irrespective of its mutation status. The other patient had a 9-year history of indolent disease without therapy. The intensity of the expressed IgVH clones as determined by PCR indicated that the mutated clones were dominant in 2 patients (approximately 80%). In the patient with VH3-21 expression, the unmutated clone accounted for 80% of total expressed IgVH. These data suggest that the presence of more than 1 clone should be considered when testing for IgVH mutation status and, more importantly, that evolution of a more aggressive second clone should be suspected. The presence of 2 clones—1 arising in naïve cells and the second in memory cells—suggests the possibility that the first hit occurred very early in the ontogeny of lymphocytes and that a second hit occurred at a later stage. However, the presence of naïve and memory clones in the same family raises the possibility that the naïve leukemic cells might still go through the germinal center programming process.
The VH3-21 Gene Status Correlates with Elevated β2-Microglobulin Serum Levels and Shorter Overall Survival of Patients with Chronic Lymphocytic Leukemia.
Authors: Lech-Maranda Ewa , Robak Tadeusz , Szmigielska-Kaplon Anna , Makuch-Lasica Hanna , Solarska Iwona , Jamroziak Krzysztof , Balcerczak Ewa , Mirowski Marek , Majewski Miroslaw P. , Warzocha Krzysztof 
Published: April 2020

Abstract
Introduction: The prognostic role of immunoglobulin variable heavy chain (IgVH) region somatic hypermutation status in B-cell chronic lymphocytic leukemia (B-CLL) is now well established. However, an overrepresentation of the VH3-21 gene segment has been recently reported to influence clinical course of B-CLL regardless of the IgVH mutational status in different geographic populations. It might suggest the role of a common antigen epitope in the pathogenesis of some B-CLL cases.
Aims: We therefore investigated the frequency of the VH3-21 gene usage in IgVH rearrangements in Polish population of B-CLL patients, and assessed whether it might influence B-CLL clinical characteristics and prognosis.
Patients and Methods: The study comprised of 97 B-CLL patients treated at the Department of Hematology, Medical University of Lodz in Poland. Peripheral blood mononuclear cells were separated on Ficoll and DNA was isolated using column method. The monoclonal population of lymphocytes with VH3-21-JH rearrangement was tested using framework 3 region primer and consensus JH primer described by Davies et al. (Leukemia 2004, 18: 872). The PCR reactions were followed with heteroduplex analysis of PCR products and separation in polyacrylamide gel. The positive VH3-21-JH samples were also confirmed by sequencing.
Results: The frequency of the VH3-21 gene segment in VH-JH rearangements in B-CLL patients reached 9% in Polish population. Beyond an elevated β2-microglobulin serum levels (p=0.03, Mann-Whitney test), no association were found between the VH3-21 gene status and clinical characteristics including CD38 expression at diagnosis The mortality rate was significantly higher in patients carrying VH3-21 (p=0.03, χ2 test), and majority of deaths were related to CLL. With a median follow-up of the surviving patients of 55,5 months (range 2–209 months), the subgroup of patients with the VH3-21 had significantly shorter overall survival (p=0.025, log-rank test). However, this genetic marker was not associated with freedom from progression survival.
Conclusions: We conclude that the frequency of the VH3-21 gene segment in VH-JH rearrangements in Polish population is similar to those previously published for British and Belgian populations and seems to be lower than in Mediterranean area populations (Blood2005, 15: 1678). The presence of VH3-21 gene segment correlates with increased β2-microglobulin levels at presentation and predicts shorter overall survival of B-CLL patients.
Complex Karyotype in Patients with Chronic Lymphocytic Leukemia.
Authors: Yañez Lucrecia , Cuadrado Maria Angeles , Insunza Andres , Mesones Belen Gonzalez , Bermudez Arancha , Conde Eulogio , Bureo Encarna , Iriondo Arturo 
Published: April 2020

Abstract
Chronic Lymphocytic Leukemia (CLL) is heterogeneous and there are two clinical forms with different outcome, in which initial stage and parameters of tumor burden, expression of CD38 and /or ZAP 70, chromosome abnormalities such as 11q23 and 17p13 are associated with a more aggressive course of disease.
The aim of this study is to analyse clinical and biological characteristics and outcome of patients with complex karyotype, and to asses if there are differences with other genomic aberrations.
Fifty patients diagnosed of CLL between 1980 and 1994 were studied (median age: 71 yrs; female/male: 18/32; median follow up 6.6 yrs.). Biological studies with flow cytometry and fluorescence in situ hybridization were done on cryopreserved blood cells. Clinical stages at diagnosis were: Rai stage 0: 56%; I: 20%; II: 16%; III: 6%; IV: 2%. Binet A: 72%; B: 20%; C: 8%. At the time of diagnosis 37% of patients needed treatment and 61% presented disease progression. At the time of this study, 10% of patients are alive with a median follow up of 15.3 yrs.
Seven patients (28%) presented complex karyotype. At the time of diagnosis 43% patients needed treatment and clinical stages were: Rai stage O: 29%, I: 14%, II: 43%, III: 14%, IV: 0%. Binet A: 43%; B: 43%; C: 14%. Median overall median survival was 72 months.
The most frequent genomic aberrations associated with complex karyotype were: del (17p13) and trisomy 12 (60 %). Cellular expression of CD38 and ZAP 70 was not able to separate this group.
Compared with other chromosome abnormalities, there were not important differences in clinical stage at diagnosis or expression of CD38 or ZAP 70, but overall median survival could distinguish three groups: del 11q22-23 or del 17p13 (34 months, p &lt; 0,05) complex karyotype (72 months), and trisomy 12 or del 13q14 (79 months, p &gt; 0,05).
In conclusion, complex karyotype is frequently formed by del 17p13 and trisomy 12 and identifies a subgroup of patients with better prognosis compared with isolated p53 deletion. More studies have to be realized to determine if trisomy 12 is a cell defence mechanism.
Involvement of PKC-δ in the Regulation of Notch2 in B-CLL.
Authors: Hubmann Rainer , Duechler Markus , Hilgarth Martin , Schnabl Susanne , Schwarzmeier Josef D. , Jaeger Ulrich , Shehata Medhat 
Published: April 2020

Abstract
We have recently shown that NOTCH2 signaling is involved in the overexpression of CD23 in B-cell chronic lymphocytic leukemia (B-CLL) cells (Hubmann et al., BLOOD 2002). There is an increasing evidence that NOTCH2 plays a determining role in the development/homeostasis of self-reactive CD5+ B-cells, suggesting a potential function of NOTCH2 in B-cell leukemogenesis. Here we study the regulation of NOTCH2 signaling in B-CLL cells and its possible function in B-lymphocytes using NOTCH2 transduced BL41 cells as a model system. Cultured B-CLL samples (n=30) lose their nuclear NOTCH2 (N2IC) activity within one day as demonstrated by electrophoretic mobility shift assays (EMSA). However, DNA-bound NOTCH2 complexes could be maintained in culture by exposure to the phorbolester TPA and is accompained by increased cell viability. The effect of TPA is prevented by the PKC-δ inhibitor Rottlerin indicating that PKC-δ is involved in the regulation of NOTCH2 signaling in B-CLL cells. The activity of N2IC in the leukemic cells appeared to be resistant to the γ-secretase inhibitors DAPT and compound E, two substances known to block ligand mediated release of the NOTCH2 intracellular domain (N2IC). Since B-CLL cells are locked in an anergic state, we next asked whether NOTCH2 modulates B-cell receptor (BCR) signaling and found that retrovirally transduced N2IC rescues the B-cell line BL41 from surface immunoglobulin M (sIgM) mediated apoptosis, a mechanism proposed to prevent the uncontrolled expansion of self-reactive CD5+ B-cells. In summary, our data suggest that B-CLL cells express an activated form of NOTCH2 which might be involved in the protection of the malignant clone from peripheral negative selection.
Interphase Fluorescence In Situ Hybridization Detection of Cytogenetic Abnormalities in B-Cell Chronic Lymphocytic Leukemia.
Authors: Xu Wei , Li Jianyong , Pan Jinlan , Li Li , Qiu Hairong , Xue Yongquan , Ruan Changgeng 
Published: April 2020

Abstract
The most frequent chromosomal abnormalities in B-cell chronic lymphocytic leukaemia (B-CLL) are deletions on 13q14 and 17p13, trisomy 12 and 14q32 rearrangement. Conventional metaphase cytogenetic analysis underestimates the frequency of specific chromosome aberrations in B-CLL due to the low rate of spontaneous mitoses and poor response to mitogen stimulation. The aim of this study was to investigate the incidence of chromosomal changes in bone marrow or peripheral blood cells (or both) of B-CLL patients using a molecular cytogenetic method, interphase fluorescence in situ hybridization (I-FISH). Probes for 13q14 (D13S319), 17p13 (P53 gene), the centromere of chromosome 12 (D12Z3) and 14q32 (Ig10 and Y6) were applied to detect chromosomal aberrations on bone marrow and peripheral blood smears from 83 B-CLL patients (60 male, 23 female,). Molecular cytogenetic aberrations were found in 60 (72.3%) cases, and 8 (9.6%) patients showed two kinds of abnormalities. The most frequent abnormalities detected in our patients was deletions of 13q14 in 34 cases (41.0%), followed by trisomy of chromosome 12 in 16 patients (19.3%), deletions of 17p13 in 10 patients (12%) and 14q32 rearrangement in 8 patients (9.6%). Statistical analyses were performed to correlate the molecular cytogenetic findings with Binet stages. No apparent differences in distribution were noted for anomalies del(13q14), del(17p13), +12 or 14q32 rearrangement among patients with various Binet stages. FISH was found to be a more rapid, exact and sensitive technique for the analysis of chromosome aberrations in CLL. FISH could provide accurate information of molecular cytogenetics for CLL.
Quantitative RT-PCR Assessment of Cannabinoid Receptor CB2 and Fatty Acid Amine Hydrolase in Neoplastic Cells of Patients with B-Cell Chronic Lymphocytic Leukaemia.
Authors: Piszcz Jaroslaw A. , Kloczko Janusz S. , Ziarko Slawomir , Radziwon Piotr , Mazgajska-Barczyk Katarzyna Z. 
Published: April 2020

Abstract
Background:
Chronic lymphocytic leukaemia (CLL) is one of the diseases in which processes of proliferation and apoptosis are seriously altered. Endocannabinoid system can modulate these processes. Cannabinoid receptor CB2 mainly expressed on lymphocytes and fatty acid amine hydrolase (FAAH) constitute a part of endocannabinoid system responsible respectively for interaction and degradation of cannabinoids. Both the receptor and enzyme are membrane proteins that can influence the lymphocyte fate. It has recently been suggested that this system may be a new potential target of anticancer therapy.
Aims:
The aim of our study was the assessment of receptor CB2 and FAAH mRNA expression in the lymphocytes B derived from patients with newly diagnosed chronic lymphocytic leukaemia.
Material and methods:
We assessed 15 peripheral blood samples from the CLL patients and 5 samples from healthy donors as a control group. The expression of CB2 receptor and FAAH proteins were measured indirectly by determination of their specific mRNA. The isolation of mononuclear cells from peripheral blood was carried out with the use of density centrifugation method. The resolution of mononuclear cell population to lymphocytes B subpopulation was done with negative isolation method utilising magnetic microballs. Total RNA was isolated from B-lymphatic cells of which 10ng was used in each reaction. RT-PCR was carried out with the use of Light Cycler (Roche) and respective commercial kits. The nucleotides sequence and parameters of the amplification process were based on method previously described by Matias et al (2002).
Results:
We found significantly higher mRNA levels of CB2 receptor in the study group in comparison to the control group. The levels of FAAH mRNA were also higher in the study group in comparison to the controls. Furthermore the number of mRNA copies was higher in case of CB2 receptor than FAAH in the CLL patients. Those numbers were comparable in the healthy controls. The results are presented as a mean number of copies in figure below.
Conclusions:
The results of our study show that during the leukaemic transformation there might be an alteration in the expression of some constituents of the endocannabinoid system. Those findings suggest the potential role of the endocannabinoid system in the neoplastic processes. Thus the assessment of the CB2 and FAAH mRNA might help to understand the biological reactions underlying the complex etiology of neoplastic disorders. Furthermore the explanation of all aspects of the role of endocannabinoid system in lymphoid malignancies should help to make it a new target of antileukaemic treatment.
figure1 figure1.
Comparison of IgH Gene Rearrangement Configuration between Hairy Cell Leukemia (HCL) and Hairy Cell Leukemia Variant (HCL-v).
Authors: Gonzalez David , Morilla Ricardo , Morilla Alison , Else Monica , Osuji Nnenna , Catovsky Daniel , Dearden Claire , Morgan Gareth J. , Matutes Estella 
Published: April 2020

Abstract
Hairy cell leukemia (HCL) is a rare B-cell disorder, with a variant form, which differs from the former in clinical behaviour, morphology and immunophenotype. The resemblance of these malignancies to splenic marginal zone lymphoma (SMZL), has led to the suggestion that they may share a common clonal progenitor that homes within the marginal zone of the splenic germinal centres. The immunophenotype and recent global gene expression results offer some support for this view, with a similar profile seen in HCL, and a subset of memory B-cells that are localized to the marginal zone. However, studies of the mutational status of the Ig genes are controversial. The majority of HCL cases are mutated, whereas up to 30% of SMZL cases are unmutated. Further investigation to understand these differences and to identify the cell of origin in these disorders is needed. In addition, there are no data on gene expression or VH mutation in HCL-variant (HCL-v), an entity with a poorer outcome and for which there is as yet no effective therapy.
We have analysed a series of 13 HCL and 23 HCL-v, for the configuration of the Ig heavy chain rearrangements. VDJH rearrangements were PCR-amplified and sequenced using an automated 3130xL sequencer (Applied Biosystems, Warrington, UK). The sequences obtained were compared to the closest germline segments using V-base (CRI, Cambridge, UK) in order to study the level of somatic hypermutation as well as gene segment usage in IgH rearrangements.
The majority of the HCL (92%) showed more than 2% deviation from the closest germline VH gene segment, compared to only 17/23 (74%) in the HCL-v cases. In the mutated cases, the percentage of mutation was similar for HCL and HCL-v (mean 5.8% and 5.9%, respectively), and none of the cases appeared to show ongoing hypermutation. Within the HCL-v group, 4 out of the 6 unmutated cases used VH4-34 segment, and the remaining 2 used VH1-03. Both genes are known to be associated with autoimmune diseases, and they are excluded from the normal and malignant memory plasma cell repertoires. Also, JH5 segment was found in 47% of the HCL-v, while it was absent in the HCL repertoire. This finding is particularly interesting, as JH5 segment is normally under-represented in most normal and malignant B-cells, except for a characteristic group of IgG-expressing chronic lymphocytic leukemias. Our data suggest that HCL-v is, unlike HCL, heterogeneous regarding IgVH mutations, but has restricted IgH repertoire that, at least in some cases, might be derived from activated, self-reactive B-lymphocytes that have not experienced the germinal-centre reaction.
Symptom Assessment in Patients with Chronic Lymphocytic Leukemia: Validation and Application of the M.D. Anderson Symptom Inventory.
Authors: Wang Xin Shelley , Ferrajoli Alessandra , Broadway Jackie B. , Mobley Gary M. , Mendoza Tito R. , Keating Michael J. , Cleeland Charles 
Published: April 2020

Abstract
CLL is the most prevalent form of adult leukemia. The life expectancy of patients affected by CLL varies significantly, with a large fraction of patients with survival expectation of 10 years or longer. Furthermore a number of patients are observed without treatment intervention. These patients have been labeled as “asymptomatic” and very little attention has been placed to the symptom burden that accompanies CLL. In order to measure the quality of life of patients with CLL, we explored the reliability and validity of the M. D. Anderson Symptom Inventory (MDASI). We enrolled 99 consecutive patients followed at our outpatient center. To assess and construct validity, we performed principal axis factor analysis with oblimin rotation. To assess reliability, we calculated separate coefficient alphas for the severity and interference subscale items. Patients were 63% male with a mean age of 59 years (range 29–79), with 43% age 60 years or older. 94% were White Non-Hispanic patients. 61% were employed. 83% had higher than high school education. 75% had Rai staging 0–1, and 53% had received prior treatment. Factor analysis of the MDASI items resulted in a two-factor solution (symptom severity and interferences) that satisfied criteria for interpretability and model fit in a confirmatory setting. Cronbach coefficient alphas for the MDASI were 0.91 for severity and 0.96 for interference. We also examined the criterion validity of the two MDASI subscales by investigating their correlation to similar subscales of the SF-12. MDASI “worst pain” was highly correlated with the “body pain” subscale of the SF-12 (r=−.84, p=.000). MDASI “fatigue” was significantly correlated with the SF-12 “energy” item (r=.79, p=.000). The patient self reported most severe symptoms were fatigue, sleeping disturbance, drowsiness, distress, and shortness of breath. There were no significant differences of symptom severity by age group (more or less than 60 years old) and by staging (Rai 0–1, 2–3). However, fatigue severity was significantly higher in patients who had more than 2 therapies than patients without prior therapy (4.26 vs 2.07 on 0 to 10 fatigue scale on MDASI, P=.003). The most burdensome symptoms were interference of working, activity and enjoyment of life. Moderate to severe symptoms that would be the targeted area of symptom control in CLL patients were fatigue (25%), drowsness, (13%), shortness of breath (11%), and sleeping disturbance (12%). In conclusion this preliminary data analysis showed that the MDASI is a reliable and valid tool for assessing symptom severity and interference with daily functioning in CLL patients. This versatile assessment tool can be used to measure CLL-related symptom burden in the clinical practice of symptom management as well as to measure changes in symptom burden in patients enrolled in therapeutic clinical trials.