bluuebunny/crossref_metadata_2025_split huggingface dataset:
Follow-Up of Clinical Manifestations and Platelet Function Test in Patients with Essential Thrombocythemia on Anagrelide Treatment.
Authors: Vassallu Patricia S. , Goette Nora P. , Glembotsky Ana C. , Heller Paula G. , Kornblihtt Laura I. , Lev Paola R. , Chazarreta Daniel C. , Salim Juan P. , Laguna Maria S. , Marta Rosana F. , Molinas Felisa C. 
Published: April 2020

Abstract
Introduccion: Patients with essential thrombocythemia (ET) may have abnormalities of platelet function (PF), the most frecuently are reduced response to epinephrine and ADP-induced aggregation and spontaneous aggregation. Anagrelide (A) has a phosphodiesterase inhibitor activity and may inhibit PF. Therefore, we evaluated the follow-up of clinical manifestations (CM) and the PF in a group of 42 ET patients before treatment and on hematological remission.
Materials and methods: 42 ET patients (33/9, F/M) who meet de PVSG criteria, were studied before treatment with A and on hematological remission. The CM were grouped into thrombosis (T), hemorrhages (H), microcirculation disturbances (MD) and combined (C), that included H plus MD or H plus T. Platelet function tests were perfomed with different agonists such as epinephrine, ADP, ristocetine, arachidonic acid and bovine vWF.
Results: are shown in Table I.
Abbreviations: T≠ (4 TIAs, 1 stroke, 1 AMI), H* (1 upper gastrointestinal bleeding, 4 minor hemorrhages) C± (1 mesenteric, portal and splenic vein thrombosis plus minor hemorrhage, 4 minor hemorrhages plus MCD), Tψ (1 TIA, 1 stroke), H§ (1 lower gastrointestinal bleeding) C† (1 AIT plus minor hemorrhage).
During hematological remission spontaneous platelet aggregation disappeared in all patients except one. Abnormal epinephrine response desappeared in 16 patients, was not corrected in 10 and this abnormality appeared in 2 whose response were previously normal. Abnormal ADP response disappeared in 4 cases during treatment and this abnormality was found in 5 previously normal cases. No correlation was found between spontaneous platelet aggregation and clinical manifestations p= NS. No correlation was found between both epinephrine and ADP response and hemorrhages. No significant abnormalities were found in the response to arachidonic acid, ristocetin, and bovine vWF before or during treatment with anagrelide.
Conclusions: In this group of ET patients platelet function was not affected by the phosphodiesterase inhibibitor activity of anagrelide. Similarly bleeding manifestations were not worsen during this therapy. As previously shown spontaneous platelet aggregation desappeared in all but one patient. We have not yet an explanation for the increased microvascular disturbances during treatment.
Table I Pre treatment On treatment Clinical Manifestations (42 patients ) Asymptomatic 16 18 Thrombosis 6≠ 2ψ Hemorrhages 5* 4§ Microvascular disturbances 10 17 Combined 5± 1† Platelet function Spontaneous aggregation 17/42 1/41 Abnormal epinephrine response 26/41 12/42 Abnormal ADP response 17/41 9/41
Function of CD39 (NTPDase-1) in Chronic Lymphocytic Leukemia.
Authors: Pulte E. Dianne , Olson Kim , Islam Naziba , Broekman M. Johan , Ballard Harold , Marcus Aaron J. 
Published: April 2020

Abstract
Chronic lymphocytic leukemia (CLL) is characterized by accumulation of large numbers of mature lymphocytes. However, thrombotic events in CLL are relatively rare. CD39, the main control system for blood fluidity, was identified on the surface of CLL cells in about 80% of patients in two recent studies. This suggests that CD39 could play a role in prevention of thrombotic events in CLL. Lymphocytes were prepared from peripheral blood of an initial 13 patients with CLL by histopaque isolation and purified to at least 95%. The ADPase and ATPase activities in these lymphocytes was measured by our radio-thin-layer chromatographic assay. Additionally, cells were stained with antibodies to CD3, CD19, and CD39, and analyzed by FACS. Remaining lymphocytes were pelleted and frozen for semi-quantitative RT-PCR analysis of the CD39 transcipt. Lymphocytes from patients with CLL showed increased ATPase and ADPase activities compared with lymphocytes from normal donors. Average ADPase activity per 50,000 cells from CLL patients was 57pmol/min, compared with 14pmol/min in lymphocytes from normal donors. Average ATPase activity was 43.6pmol/min in CLL patients, and 10.6 pmol/min in normal lymphocytes. Semi-quantitative RT-PCR of CLL lymphocytes showed upregulation of CD39 RNA compared with normal lymphocytes. FACS data showed that the percentage of cells expressing CD39 was greater in CLL cells (62.5%) than in normal lymphocytes (11%). Additionally, some CLL cells appear to express CD39 at higher densities than normal lymphocytes. Interestingly, one patient in whom lymphocytes were isolated before and during an accelerated phase of the disease showed a decrease in CD39 expression on his lymphocytes as the disease progressed. Two other patients with advanced disease, requiring chemotherapy, showed relatively low CD39 levels as well. We conclude that CD39 is present and functions as an ecto-nucleotidase on the surface of most malignant lymphocytes in CLL. The quantities are much higher than observed in normal lymphocytes. CD39 may be protective from thrombotic events in patients with CLL. In addition, lymphocyte CLL may be a marker of earlier or less aggressive disease and a decrease of CD39 on the cell surface may signify a worsening of the disease.
Toll like Receptors (TLRs) in B Cell Chronic Lymphocytic Leukemia (B-CLL).
Authors: Manoussaka Maria S. , Memon Azka , Nathwani Amit , Porakishvili Nino , Clark Edward , Lydyard Peter M. 
Published: April 2020

Abstract
Introduction: B-CLL is characterised by the accumulation of long-lived CD5+ B cells. Mature B cells can be activated via TLRs to produce immunoglobulin and proliferate. We have previously reported differential surface expression of CD180, TLR-4 associated receptor, by B-CLL cells in relation to IgVh gene status (Porakishvili N et al., 2004; Blood (ASH annual meeting abstracts) 104: 2807). To further characterise subsets of leukemic cells, we assessed B-CLL cells for TLR mRNA expression.
Method: B cells from untreated (n=24) and treated (CHOP n=3, Thalidomide n=1 and chlorambucil and prednisolone n=2) CLL patients, and healthy aged matched controls, (n=14) were isolated by CD19+ selection using iMACs columns (Miltenyi Biotec), total RNA extracted (Qiagen) and primers for TLR2, TLR4, TLR9, CD180 and the adaptor protein MYD88 were used for TaqMan PCR analysis (ABI). GAPDH was used for normalization. Data were expressed as mean delta cycle number (ct) ± standard deviation, giving a value inversely related to the level of expression of the specific gene. Patients were separated into groups according to IgVh gene mutation status or level of Zap 70 expression. Statistical analysis was performed using by Mann-Whitney U test and Spearman rank correlation or Pearson’s correlation coefficient.
Results: Overall, there was a heterogeneous pattern of TLR expression in patients and controls with no significant differences between them. Treated patients were characterised by higher mRNA levels for the all TLRs measured (TLR9 p=0.018) and MYD88 (MYD88 p=0.026) compared with untreated patients, despite no significant difference of their GAPDH levels (p&gt;0.05). Significant correlations were seen between mRNA levels for the TLRs within the age matched control group (all p&lt;0.05). Groups of patients were further analysed according to mutated (mutated &gt;2%) or unmutated IgVh genes (unmutated &lt;2%). No significant correlation in the TLR expression levels was found within the mutated patient group (n=8) or Zap 70 low expression group (&lt;0.7 level) (n=11). Preliminary data from patients with an unmutated IgVh gene status (n=4) or according to levels of ZAP 70high expression (n=5) showed correlation of mRNA levels for TLRs and MYD88 expression (all p&lt;0.05).
Conclusion: Our data indicates a direct relationship between MYD88 and TLRs transcription levels in healthy control individuals. Lack of significant correlation amongst TLR mRNA levels in CLL patients with mutated IgVh genes and Zap70low expression levels suggests aberrant regulation of TLRs in this group.
Characteristics and Outcome of B-Cell Chronic Lymphocytic Leukemia in HCV-Positive Patients.
Authors: Molica Stefano , Mirabelli Rosanna , Misuraca Demetrio , Battaglia Caterina 
Published: April 2020

Abstract
HCV-associated B-cell non-Hodgkin’s lymphomas (NHL) show distinctive clinico-pathological features such as older age, liver damage, presence of monoclonal gammopathy, increased incidence of autoimmune disorders, extranodal localizations and restricted histological subtypes. As far as B-cell chronic lymphocytic leukemia (CLL) is concerned, information dealing with either characteristics or outcome of HCV-associated CLL are limited. With this background we compared clinico-hematological features and outcome of 34 HCV-positive patients diagnosed at our institution as having immunologically typical B-cell CLL (i.e., CD5+/CD23+/CD79b-/SmIg dim) with 161 unselected CLL HCV-negative patients followed-up in the last 10 years. The two groups were alike with respect to main clinico-hematological features such as age (P=0.780), sex (P=0.650), absolute lymphocyte count (P=0.788), platelet count (P=0.362), haemoglobin level (P=0.704), β2-microglobulin (P=0.192), Binet stage distribution (P=0.224) and lymphocyte doubling time (LDT)(P= 0.620). As expected either ALT or AST serum levels at the time of CLL diagnosis were significantly higher in HCV-positive patients in comparison to HCV-negative ones (P&lt;0.0001 for both). In contrast, no difference was found in the incidence of monoclonal gammopathy between HCV-positive and HCV-negative patients (10.3% versus 7.7%; P=0.708). The same applied for autoimmune disorders which were homogeneously distributed in the two subgroups (P=0.711) and accounted, more frequently, for autoimmune emolytic anemia (AEA)(HCV-negative subgroup, 5.5%; HCV-positive subgroup, 9.0%). The proportion of severe infections registered did not reflect the HCV-status (HCV-negative subgroup, 9.6%; HCV-positive subgroup 6.4%; P= 0.510). Also second tumours were equally distributed among HCV-positive and HCV-negative subgroups (10% versus 6.8%; P=0.655). Survival curves projected at 10 years did not show any statistical in terms overall survival (Hazard Risk, 0.690; 95% CI: 0.216–1.304; P=0.167). Finally, the short term hepatic toxicity of chemotherapy did not increase among HCV-positive patients (P=0.671).
In conclusion, HCV-positive patients with B-cell CLL do not differ from other patients both for presentation and clinical outcome. The need to activate specific protocols of antiviral therapy appears less urgent in comparison to NHL, however, younger CLL patients HCV-positive who are eligible for therapies at higher immunosuppressive potential (i.e., chemo-immunotherapy) should be taken in special consideration.
Polymorphisms in the Promoter Region of the Mcl-1 Gene Are Associated with Ex Vivo Cytotoxicity but Not with Response to Treatment in Advanced Chronic Lymphocytic Leukemia (CLL).
Authors: Villamor Neus , Marce Silvia , Bosch Francesc , Ferrer Anna , Pérez-Galán Patricia , López-Guillermo Armando , Giné Eva , Campo Elías , Montserrat Emili , Colomer Dolors 
Published: April 2020

Abstract
High levels of Mcl-1 have been related to a poor response to therapy in patients with CLL. Genetic polymorphisms in the promoter region of the Mcl-1 gene consisting in insertions of 6 (+6) or 18 (+18) bp have been described in 30% to 60% of patients with CLL. The biological and clinical translation of such polymorphisms is controversial. We have analysed this issue in a cohort of newly diagnosed patients with CLL (14 F/ 46 M; 80% in B or C Binet’s stage), treated with a combination of fludarabine (F), cyclophospamide (C) and mitoxantrone (M). Polymorphisms were correlated with:
Mcl-1 RNA expression as assessed by quantitative PCR, ex vivo cytotoxicity against F, C, and M, alone or in combination, using the MTT assay, and clinical response to treatment.
The frequency of the wild type (wt), +6, and +18 allel was 56%, 12%, and 32 %, respectively (table). Twenty-one of 60 (35%) patients were homozygous for wt allel. Quantification of Mcl-1 RNA expression was performed in 18 patients. Wt/wt patients (n=6) tended to have lower Mcl-1 gene expression than the remaining patients (1.0 AU ±0.4 vs 2.2 AU ±2.8). Ex vivo cytotoxicity was different according to the Mcl-1 genotypes for F, M, F+M, and C+F, but not for FCM. The highest cytotoxicity was observed in patients with +18/+18 genotype (n=7) while those with wt/wt (n=11) presented intermediate cytotoxicity and those with other genotypes (n=17) had the lowest cytotoxicity. Twenty-eight patients achieved CR, 22 PR, and 5 failed to respond. No relationship was found between Mcl-1 polymorphisms and response to therapy (table). Interestingly, however, the ex vivo response to F (58.2% ±16 vs 45.7% ±16, p=0.05) and F+M (87% ±9 vs 76% ±15.5, p=0.04) was greater in the 13 patients who achieved MRD-negative CR. After a median follow up of 28 months (range: 6–53), 16 of 38 patients have progressed. No relationship was found between Mcl-1 polymorphism and progression after treatment. In summary, in this series of patients with advanced CLL homogeneously treated the frequency of insertions in the promoter region of Mcl-1 gene was high. Although Mcl-1 polymorphisms were associated with the ex vivo response to antileukemic drugs, they did not correlate with response to treatment and disease progression.
Distribution of Mcl-1 Polymorphisms and Response to Treatment Allel Allelic Frequency Genotype Patients CR (MRD- CR) wt/wt 21 (35%) 9 ( 4 ) wt 68 (56%) +6/+6 2 ( 3%) 0 ( 0 ) +18/+18 7 (12%) 4 ( 3 ) +6 14 (12%) wt/+6 6 (10%) 4 ( 2 ) wt/+18 20 (33%) 9 ( 3 ) +18 38 (32%) +6/+18 4 ( 7%) 2 ( 1 )
Aggressive Cutaneous Squamous Cell Carcinoma (SCC) in Chronic Lymphocytic Leukaemia (CLL).
Authors: Lane Steven W. , Mollee Peter N. , Bird Robert , Porceddu Sandro , Gill Devinder 
Published: April 2020

Abstract
Aim
The aggressive nature of cutaneous SCC in CLL, specifically presentation with locally invasive, regional metastatic or loco-regionally recurrent disease, is not well documented. We describe a series of patients with CLL who have developed aggressive SCC.
Methods
Patients with CLL with aggressive SCC were identified from existing databases in the Department of Haematology and Oncology, Princess Alexandra Hospital, Australia from August 2001–May 2005. Patient demographics, tumour characteristics and treatment received were reviewed from the clinical records.
Results
We identified 12 patients with CLL with aggressive head and neck SCC: 11 locally invasive, 7 with lymph node metastases and 5 with loco-regional recurrence.
The median age was 73 (range 49–88) and 9 patients were male. 8 patients were Rai stage III or IV. 8 of 9 patients with documented IgG levels had hypogammaglobulinemia (median 3.4g/L). 143 patients with CLL were seen in the hematology department during this time. The median time from CLL diagnosis to SCC development was 55 months (range 0–169). Median time from chemotherapy to aggressive SCC was 48 months (range 22–110) with alkylating agent chemotherapy (seven patients) and 12.5 months (range 1–46) with purine analogue chemotherapy (six patients). Four patients had received no treatment for CLL. All patients had extensive surgical resection and 11 had, or are planned for, post-operative radiotherapy. Five developed loco-regional recurrence at a median of 12 months (range 4–14). 10 patients had numerous SCC (&gt;5) excised prior to the diagnosis of aggressive SCC.
Conclusions
The risk of patients with CLL developing aggressive cutaneous SCC, with its attendent morbidity, is underappreciated. Cellular and humoral immunodeficiencies from CLL and its chemotherapy treatment may predispose patients to aggressive SCC.
The Molecular Basis for BCL-2 Oncogene Addiction in CLL.
Authors: Letai Anthony , Certo Michael , Brown Jennifer R. , Moore Victoria 
Published: April 2020

Abstract
CLL cells consistently express BCL-2 at a high level. Using a compound demonstrated to antagonize BCL-2 function, ABT-737, we show that CLL cells in short-term primary culture are uniformly sensitive to single-agent BCL-2 antagonism, with EC50’s in the 10 nM range. To understand the mechanism of CLL sensitivity to this compound, we studied mitochondria from these primary CLL cells. We employed a panel of peptides derived from the BH3 domain of pro-death BH3-only proteins, certain of which selectively inhibit BCL-2 function in vitro. We demonstrate that those with activity against BCL-2 in vitro induce cytochrome c release, a hallmark of the mitochondrial dysfunction during apoptosis. A non-malignant cellular model of BCL-2 dependence revealed that binding of the pro-apoptotic protein BIM to BCL-2 correlated with sensitivity to BCL-2 inhibitors. We likewise discovered that BCL-2 binds and sequesters BIM in CLL cells. In contrast to the BCL-2 dependent CLL cells, we examined myeloma cell lines with defined MCL-1 dependence and relative BCL-2 independence. These cell lines were 100 fold less sensitive to ABT-737. Furthermore, in mitochondrial assays only BH3 peptides which interact with MCL-1 induce cytochrome c release. These experiments reveal that cellular requirement for an anti-apoptotic protein function can be deduced from a functional mitochondrial requirement determined using our panel of BH3 peptides. In CLL, and perhaps other malignancies, it appears that BCL-2 maintains survival by sequestering pro-apoptotic proteins like BIM. Inhibition of this sequestration provokes cell death ex vivo, even with a single agent BCL-2 inhibitor. Reliance on BCL-2 for survival may be an Achilles heel for CLL and other cancers.
Hypercalcemia in B-Cell Chronic Lymphocytic Leukemia: Report of a Case and Review of the Literature.
Authors: Radhakrishnan Neetu , Hoffman Mark A. 
Published: April 2020

Abstract
A 53 year old man with Rai stage IV CLL was being treated with R-HyperCVAD when he presented between cycles with fatigue, lethargy and pancytopenia. Clinical examination revealed diffuse adenopathy and splenomegaly. Serum calcium was 13.6 mg/dl, phosphorus level was 2.7 mg/dl, and alkaline phosphatase was 54 U/L. PTH was 4 Units/L (10–65), PTHrP &lt; 0.2 pmole/L (0–1.9), 1–25 (OH) Vitamin D &lt; 10pgm/ml (22–67). Quantitative immunglobulins: IgA 10 mg/dl, IgG 252 mg/dl, IgM 50 mg/dl. Immunofixation revealed a faint IgG lambda paraprotein. There were no lytic lesions on skeletal survey. Bone marrow biopsy revealed focal large cell transformation (Richter’s syndrome). Cytogenetics revealed 3 metaphases with complex cytogenetic abnormalities, indicating clonal evolution. The hypercalcemia resolved with appropriate therapy, but despite subsequent treatment with CAMPATH, he died 2 weeks after diagnosis.
A review of reported patients with CLL and hypercalcemia in the literature was performed from 1980 onwards using MEDLINE and PubMed; only those cases in which clinical aspects, biochemistry, PTH levels, imaging studies and concurrent pathology (if obtained) were documented, are summarized in this analysis (n=13).
Rai stage: I n=1, II n=4, III n=3, IV n=5. Immunoreactive PTH levels were low or normal in 100% of patients. In 5 cases in which it was measured, 1–25 (OH) Vitamin D levels were not elevated. PTHrP was normal in 2 cases and elevated in 1. In nine patients, multiple lytic bone lesions were present on skeletal radiology. Two patients had osteopenia without lytic lesions. Two had no lytic lesions.
Six of ten patients had evidence of transfomation on lymph node and/or bone marrow biopsy performed at the time of evaluation for hypercalcemia.
Prognosis was poor (range 0.5–12 months) with only one patient surviving post allo-transplant.
In conclusion, hypercalcemia in CLL is rare. Osteolytic lesions are present in the majority of cases. PTH levels are low, and thus this hormone is not mediating the hypercalcemia. The evidence is also against a role of elevated Vitamin D. Histological transformation is seen in half of the cases. Survival is poor after diagnosis of hypercalcemia. The mechanisms(s) of the osteolysis and hypercalcemia remain to be defined.
Influence of Genetic Single Nucleotide Polymorphisms in CYP3A4, CYP3A5, GSTP1, GSTM1, GSTT1 and MDR1 Genes on Survival and Treatment Related Toxicity in Patients with Multiple Myeloma Treated in the HOVON 24 Trial.
Authors: Sonneveld Pieter , Schilthuizen Celine , Lokhorst Henk , Vellenga Edo , Raymakers Reinier , de Knegt Yvonne , Kamst Eric , van der Holt Bronno 
Published: April 2020

Abstract
Cytochromes P450 (CYP 450) and gluthatione-S-transferases (GSTs) are drug metabolizing enzymes involved in the detoxification of numerous chemotherapeutic agents.The Multidrug Resistance gene 1 (MDR1, ABCB1) is a transmembrane drug transporter present in tumor cells and mucosal epithelium. Genetic single nucleotide polymorphisms (SNP’s) in these genes are frequent and may alter the metabolism of certain anti-cancer drugs. The CYP3A4*1B (A-290G) polymorphism has not yet been proven to alter enzymatic activity. Polymorphisms in the CYP3A5*3 gene (A6986G) and the GSTP1 gene (Ile105Val) lead to decreased enzymatic activity. Homozygous deletions in the GSTM1 and GSTT1 genes lead to complete absence of the enzyme. Genetic polymorphism of MDR1 (C3435T) are frequent and associated with enhanced efflux function. We hypothesized that SNP haplotypes of these drug metabolizing enzymes may determine the interindividual differences of patients to treatment response and toxicity. We have investigated the presence of SNP’s of 211/345 previously untreated patients with multiple myeloma from whom diagnostic material was available and who were treated according to the HOVON-24 protocol, a clinical trial comparing myeloablative therapy with stem cell rescue added to intensified chemotherapy to intensified chemotherapy alone. Peripheral blood or bone marrow samples were analyzed by PCR or multiplex PCR. Patients with different haplotypes of CYP3A4, GSTP1, GSTM1, GSTT1 genes did not have significant differences of overall survival (OS), time to progression (TTP), progression free survival (PFS), event free survival (EFS), partial remission (PR), complete remission (CR) as determined by multivariate analysis with all clinical variables. Patients with mutant GSTP1(Ile/Val) had significantly more toxicity following high-dose melphalan (p=0.003). Patients with MDR1 C3435T mutation or absence of GSTP1 Ile105Val mutations had a lower probability to achieve at least a partial response(p=0.01). Patients genotyped homozygously for the mutant allele CYP3A5 gene had increased toxicity (p=0.040), improved OS (p=0.01) and PFS (p=0.04) and a decrease in TTP (p=0.03).
This study is part of an ongoing analysis and the results will be validated in the next HOVON trial. We conclude that in patients with multiple myeloma who are treated with intensive chemotherapy and stem cell transplantation, analysis of genetic polymorphisms of metabolic enzymes may improve our understanding of treatment associated toxicity and treatment outcome.