bluuebunny/crossref_metadata_2025_split huggingface dataset:
Increased Stability of Bcl-2 mRNA in B-Cell Chronic Lymphocytic Leukemia (CLL).
Authors: Kio Ebenezer A. , Pineda-Roman Mauricio , Otake Yoko , Stuart Robert K. , Fernandes Daniel J. 
Published: April 2020

Abstract
B-cell CLL is characterized by the accumulation of mononuclear B cells that are resistant to apoptosis as a result of bcl-2 oncogene overexpression. Nucleolin has recently been identified as a bcl-2 mRNA stabilizing protein that binds specifically to a 139 base AU-rich instability element (ARE) (Sengupta et al., J. Biol. Chem.279:10855–10863, 2004). Thus, studies were done to address the question whether the increased levels of bcl-2 mRNA in CLL are related to stabilization of bcl-2 mRNA by nucleolin. B cells were isolated from the blood of 9 patients with CLL and 5 normal volunteers by density gradient centrifugation followed by positive selection with CD19 immuno-magnetic microbeads. Flow cytometric analysis indicated that greater than 90% of the CLL and normal B cells were CD19 positive and CD3 negative. Western blotting revealed that cytoplasmic nucleolin and total cellular bcl-2 protein levels were elevated 18-fold (p&lt;0.001) and 5-fold (p&lt;0.001) respectively, in CLL compared to normal B cells. To directly examine the ability of nucleolin to stabilize bcl-2 mRNA, in vitro RNA decay assays were carried out using capped and polyadenylated bcl-2 mRNA transcripts. The average half-life of the bcl-2 mRNA transcript containing the ARE instability element was 12.5 min in S100 extracts of normal B cells, but was increased to 39.7 min in CLL cell extracts. Purified recombinant nucleolin (280 nM) stabilized bcl-2 mRNA when added to S100 extracts of normal B cells, but had no significant effect on bcl-2 mRNA stability in CLL cell extracts. The results are consistent with the hypothesis that resistance of CLL cells to apoptosis is related to overexpression of cytoplasmic nucleolin and increased stability of bcl-2 mRNA. These findings warrant further investigation of nucleolin as a potential therapeutic target in CLL.
Assessment of Nuclear Homogenicity in Chronic Lymphocytic Leukemia by Image Analysis.
Authors: Stein Gideon Y. , Djaldetti Meir , Zeidman Aliza , Merkel Drorit , Cohen Amos M. 
Published: April 2020

Abstract
Maturation of progenitor cells involves gradual chromatin condensation. We have shown that B-CLL cells represnet specific specrtral patterns by multipixel spectral imaging (J. Histocem. Cytohem. 46:113,1998). We further used image analysis techniques to quantify the alteration of chromatin homogenicity in lymphocytes from healthy subjects and B-CLL patients, as well as during maturation of myeloid and erythroid series, using light microscopic digital images of May-Grünwald Giemsa-stained (MGG) cells (B-CLL n=60, control n=22) and transmission electron microscopy (TEM). Nuclei were outlined by mouse cursor. Light microscopy images were converted to grayscale and designated as vector of the red, green and blue channels (sqrt(R*R+G*G+B*B)). Average color intensity (0–255), entropy (0–8), standard deviation, area diversity (0–100) and positive percent area (PPA) measurements were obtained from each nucleus. The entropy and area diversity of B-CLL lymphocyte nuclei was significantly lower than control lymphocytes (entropy 5.6±0.19 vs. 5.44±0.2, respectively, p&lt;0.0001 and area diversity 33.2±4.3 vs. 30.5±2, respectively, p=0.006). In addition, we analyzed MGG-stained and TEM myeloid and erythroid cells at various stages of differentiation (n=155). Both the myeloid and erythroid cell lines showed gradual reduced color intensity, increased entropy and area diversity. Furthermore, MGG-stained megaloblastic proerythroblasts showed significantly higher levels of entropy and area diversity as compared with control proerythroblasts (n=73). Thus, assessment of nuclear entropy and area diversity may help to in the differentiation between normal lymphocytes and B-CLL cells.
R-Flurbiprofen, a Non-Steroidal Anti-Inflammatory Agent (NSAID) with Anti-Tumor Activity, Inhibits the Stromal-Induced Upregulation of Interleukin-8 in Chronic Lymphocytic Leukemia Cells.
Authors: Lilly Michael B. , Berdeja Jesus , Chen C.S. , Bashkyrova Svetlana , Zemskova Marina 
Published: April 2020

Abstract
NSAIDs have documented activity in the prevention or treatment of human epithelial cancers. Little information exists, however, to determine if similar anti-tumor effects can occur with hematologic malignancies. We have evaluated R-flurbiprofen (RFB), an enantiomer of an arylpropionic acid NSAID with known anti-tumor effects, as a potential treatment for CLL. PBMNCs from 7 CLL patients (WBC 30,000 –104,000/mcL; all CD5+/CD19+/CD38−) were exposed in short term culture to RFB. All samples showed caspase-dependent cell death following addition of RFB, with a median LD50 of approximately 90mcg/mL, a clinically achievable concentration. Addition of PGE2 or caffeic acid did not prevent RFB-induced cell death, demonstrating that the effect was not due to cyclooxygenase or lipooxygenase modulation. To identify molecular events resulting from RFB treatment, we performed microarray anaylsis (with Affymetrix U74v2A chips) on drug-treated CLL cells. PBMNCs were initially cultured in medium alone, or co-cultured with allogeneic primary marrow stromal cells (MSC). MSC co-culture increased or decreased (by &gt;2-fold) approximately 13% of the 12,000 genes/probe sets interrogated by these microarrays (2 independent experiments), but had little effect on RFB cell kill. Approximately 35 genes (fusion of 2 independent experiments) were reproducibly up- or down-regulated by RFB in the non-adherent CLL cells cultured with MSC, but not in CLL cultured alone. Among the down-regulated genes were several chemokines and adhesion molecules. These included the gene for interleukin-8 (IL8). The expression of IL8 was markedly increased in CLL cells by co-cultivation with MSC. Furthermore, IL8 expression showed reproducible and dose-dependent down-regulation in CLL cells by RFB treatment. In contrast RFB did not affect expression of IL8 in CLL cells cultured without MSC. Intracellular IL8 was measured in CLL cells by flow cytometry; RFB markedly decreased expression of IL8 protein in addition to its effects on IL8 mRNA. IL8 has been identified as a growth/survival factor for CLL cells, and is a potent tumor-derived stimulator of angiogenesis. Furthermore, increased blood levels of IL8 have been identified as a poor prognostic sign in patients with CLL. RFB may modulate leukemia cell/MSC interactions by blocking the expression of chemokines and other pro-angiogenic mediators. RFB may play a supporting role as part of a multi-modality therapy regimen for CLL.
Deletion of 13q14 Is Independent of CD38 Expression in Chronic Lymphocytic Leukemia (CLL).
Authors: Mavromatis Blanche H. , Cheson Bruce D. , Ozdemirli Metin , Hartmann Dan-Paul , Abbasi Salah , Zafer Emre , Zantek Nicole , Cooksey Jessica , Meck Jeanne 
Published: April 2020

Abstract
Cytogenetic abnormalities detected by fluorescent in situ hybridization (FISH) can predict differences in survival in patients with CLL. Patients (pts) with the 13q14 deletion experience a longer median survival that those with the 17p13.1 deletion. Overexpression of ZAP-70 and CD38 correlate with a poor prognosis. Data on 44 pts currently enrolled on a prospective CLL prognostic study were analyzed for cytogenetic abnormalities by FISH and CD38 by flow. Pt age ranged from 25 to 85 (median 59). 25/44 patients (57%) were followed without any therapy. Zap-70 expression was measured by immunohistochemistry using a monoclonal antibody to ZAP-70 (clone 2F3.2).
Results: 22/44 (50%) pts were found to have abnormal cytogenetics. Of the 22 abnormalities, 17 had a 13q14 deletion (including one with an additional 11q22.31 deletion, and two with nullisomy 13q), five patients had trisomy 12, one patient a 17 p13.1 deletion. 8/44 patients overexpressed CD38. Of the pts with an isolated 13q deletion, 12/16 (75%) were CD38 negative and 10/17 (59%) were followed without any therapy, compared to 20/27 (74%) and 14/27 (52%) in those with other cytogenetic features respectively. Of those with other abnormal FISH findings, 4/5 pts (90%) with trisomy 12 were CD38 negative, a patient with 17p13.1 deletion and another patient with both 13q14 and 11q deletion were also CD38 positive. Of the patients with normal FISH results 19/21 (90%) were CD38 negative. ZAP-70 overexpression was noted in 22/26 pts with available data including 8/10 (80%) patients with and 13/26 (50%) patients without the 13q del.
Conclusion: The presence of the 13q14 deletion appeared to be independent of CD38 expression. Further studies to determine the association of 13q14 deletion with ZAP-70 expression are warranted, as is the correlation with clinical data.
ZAP-70 Expression in Chronic Lymphocytic Leukemia (CLL) Is Independent of Rai Stage: A Retrospective Review of 38 Patients (pts).
Authors: Mavromatis Blanche H. , Cheson Bruce D. , Zantek Nicole D. , Hartmann Dan-Paul , Kessler Craig , Cohen Philip , Gehan Edmund , Ozdemirli Metin 
Published: April 2020

Abstract
ZAP-70 is a tyrosine kinase protein involved in T cell signaling, overexpression of which is associated with a poor prognosis in CLL. Clinical information on 32 pts with CLL, and 6 pts with prolymphocytic transformation (PLT) followed at our institution was retrospectively reviewed for Rai stage, ZAP-70, CD38, FISH and response to treatment to determine the prognostic association among these factors. Ages ranged from 37 to 78 (median 59). 5/32 pts with CLL had received prior initial therapy, while 17/19 pts had not. They eventually were all treated with fludarabine based regimens either as salvage or upfront. The remaining 8/32 had not required therapy. Response to therapy was correlated with ZAP-70 expression as measured by immunohistochemistry using a monoclonal antibody to ZAP-70 (clone 2F3.2).17/32 patients (53%) were ZAP-70 positive, as were 3/6 (50%) of the patients with PLT. Of the patients with CLL who were ZAP-70 positive, 5 (30%) were stage 0-II, and 12 (70%) were stage III/IV, while18% did not require therapy. Of the 13 who were treated there were 4 (31%) CR, and 5 (38%) PR. Of the patients who were ZAP-70 negative, 50% (7/14) had stages 0-II and 50% had stage III/IV disease. 5/14(36%) were not treated. Of the 9 who were treated there were 2 CR 22%, and 7 PR 78% (p=0.01). 3 PLT patients were previously treated for CLL with fludarabine-based combinations. When they required therapy for PLT, two achieved a CR (one to rituximab/fludarabine/cytoxan_ZAP-70 positive patient, and one to R-CHOP combination_ZAP-70 negative), and another a PR (to R-EPOCH combination_ZAP-70 negative patient). Of the previously untreated PLT patients, two ZAP-70 positive patients achieved a CR (one to rituximab/fludarabine/cyclophosphamide, and one to R-CHOP). One ZAP-70 negative patient was refractory to fludarabine/rituximab combination.
Conclusion: ZAP-70 positive CLL is more likely to be refractory to treatment, while ZAP-70 negative pts achieved a higher ORR (p=0.01). Although pts who were ZAP-70 positive were more likely to have advanced stage, 30% of these pts had early stage disease. As such, ZAP-70 did not closely correlate with stage, and may be an independent prognostic factor in these pts. Response to treatment could also not be predicted by ZAP-70 expression.
ZAP-70 Number of Pts Previous Treatment Number of Pts Response Positive 16 No 10 3CR, 4PR Yes 3 1CR, 1PR Negative 14 No 7 2CR, 5PR Yes 2 2PR
Hepatitis B Virus Reactivation and Alemtuzumab Therapy.
Authors: Iannitto Emilio , Minardi Viviana , Calvaruso Giuseppina , Mulè Antonino , Ferraro Donatella , Ammatuna Emanuele , Craxì Antonio , Abbadessa Vincenzo , Di Stefano Rosellina 
Published: April 2020

Abstract
Reactivation of hepatitis B virus infection in subjects receiving cytotoxic treatment for haematological malignancies occurs in 21–53% of chronic HBsAg carriers and in an unknown number of HBsAg negative subjects harbouring occult HBV infection. Immunotherapy with alemtuzumab, a humanized monoclonal antibody against CD52 on lymphoid cells, produces deep immunosuppression. We describe two subjects with chronic lymphocytic leukaemia and occult HBV infection who developed a virological and biochemical flare of hepatitis B following immunotherapy with alemtuzumab. One of them developed a full blown hepatitis with seroreversion from anti-HBs to HBsAg after four weeks of alemtuzumab. Lamivudine (100 mg die) achieved a complete clinical recovery and HBV-DNA clearance from blood within 8 weeks. The second patient (HBsAg and HBV-DNA seronegative, anti-HBs and anti-HBc positive before treatment) was kept under prophylaxis with lamivudine up to three months after alemtuzumab. Two months after withdrawal of lamivudine, clinical and laboratory features of acute hepatitis B developed. Lamivudine therapy was restarted and obtained a prompt recovery with HBsAg and HBV-DNA clearance.
These cases suggest that alemtuzumab, as all other immune response modifiers, should be used cautiously when HBV infection is present. While a recommendation for universal pre-emptive treatment with lamivudine of all HBsAg positive patients undergoing effective immunosuppressive treatment can be made it is more difficult to establish a pattern for patients who have serological markers of previous HBV infection (anti-HBs and/or anti-HBc) and even more for those with no HBV markers. Testing for HBV-DNA in serum by highly sensitive PCR may discover up to 50% of these cases, allowing preemptive lamivudine therapy. A substantial proportion would however go undiscovered in the absence of a liver biopsy, which is clearly an unfeasible proposition as a screening procedure. Once a reactivation has been documented, nucleoside analog therapy should last for a few months after HBV-DNA clearance.
A Phase II Study of Denileukin Diftitox (ONTAKR) in Patients with Fludarabine Refractory B-Cell Chronic Lymphocytic Leukemia.
Authors: Kuriakose Philip , Turturro Francesco , Berdeja Jesus G. , Kerr Robert , Surendranathan Asha , Black Jennifer , Frankel Arthur E. 
Published: April 2020

Abstract
Therapeutic options for chronic lymphocytic leukemia (CLL) at relapse are limited because of myelosuppressive toxicity. Denileukin diftitox (ONTAK®, Ligand Pharmaceuticals) is a genetically engineered fusion protein comprising the enzymatically active domain of diphtheria toxin and the full length sequence of interleukin-2 (IL-2) targeting malignancies expressing the medium and high affinity IL-2 receptors. We designed a phase II study to evaluate the efficacy of ONTAK® in patients with fludarabine-refractory CLL, which is a follow-up to the previously published study (Frankel, et al, Clin. Cancer Res.2003; 9:3555). Denileukin diftitox was administered at a dose of 18μg/kg IV daily for 5 days every 3 weeks, for a maximum of 8 cycles. Thirteen patients have been treated so far, with 10 patients being evaluable for response (completed ≥ 3 cycles). Median age was 59 years (range 44–84), and 62% (8/13) were Rai stage III-IV, with a median of 3 prior therapies (range 1–6). The overall response was 40%, with 1 CR (10%, duration of response 5+ months) and 3 PR (30%, duration of response 3+, 3+ and 4+ months). Two responding patients (both PR) are still on study, while two (1 CR, 1 PR) were removed from study because of toxicities after 7 and 5 cycles, respectively. Four patients (40%) had progressive disease after cycles 3, 4, 4, and 7, respectively. One patient has completed four cycles and restaging studies are pending. Of the 3 patients not evaluable for response, two are still on study (having not completed 3 cycles), while one refused further treatment after 4 doses of cycle one. The grade 3/4 toxicities encountered were: neutropenia 4/13, thrombocytopenia 4/13, vascular leak syndrome 3/13, left ventricular cardiac dysfunction 1/13, hypotension 2/13, tachyarrhythmia 3/13, elevated PT 1/13, fatigue 1/13, rash 1/13, SIADH 1/13, constipation 1/13, vomiting 2/13, petechiae 1/13, transient elevation of GGT 1/13, transient elevation of AST/ALT 7/13, hyperglycemia 4/13, electrolyte imbalance 8/13, infection and/or febrile neutropenia 4/13, insomnia 1/13, visual disturbance 1/13, dyspnea 2/13, hypoxia 2/13. We conclude that denileukin diftitox has activity in CLL, with toxicities that can be managed with adequate premedication and close monitoring.
Anti-Tumour Effect of Zoledronate in Cells from B Chronic Lymphoytic Leukemia and Low-Grade Lymphoma Patients.
Authors: Vasconcellos Deborah Vidal , Silva Karina Lani , de Paula Castro Eric Delfraro , Moellman Arthur Coelho , Pombo-de-Oliveira Maria do Socorro , Maia Raquel Ciuvalschi 
Published: April 2020

Abstract
Recent studies demonstrate that bisphosphonates - anti-resorptive drugs - have direct anti-tumour effect in many tumour cell lines, including hematopoietic ones. The aim of this study was to evaluate the apoptotic effect of Zoledronate in cells from B chronic lymphocytic leukemia (B-CLL) and low-grade lymphoma (LGL) patients. Samples of 19 B-CLL or LGL patients were incubated with Zoledronate 100 μM in RPMI medium supplemented with fetal bovine serum 10% at 370C for 12h. Apoptosis using Annexin V assay, by flow cytometry (FC), was observed in 8 out of 19 (42,1%) patients despite previous treatment. Multidrug resistance (MDR) phenotype was performed using rhodamine-123 efflux assay by FC. Our results demonstrate that Zoledronate 100 m M can induce apoptosis in B malignant lymphocytes despite previous treatment and MDR phenotype. So, patients previously treated with distinct therapeutic agents or not can potentially benefit from the anti-tumour effect of Zoledronate. This is the first work demonstrating the anti-tumour effect of Zoledronate in newly obtained cells from patients with B-CLL and LGL. These results in association with evidences from recent studies suggest that Zoledronate may have an important and complementary role in hematological malignant diseases.
SPAN-Xb Expression in Myeloma Cells Can Be Upregulated by Pharmacologics.
Authors: Wang Zhiqing , Zhang Yana , Zhang Jian , Lim Seah H. 
Published: April 2020

Abstract
SPAN-Xb is a spermatid-specific protein. We recently identified SPAN-Xb as a novel Cancer-Testis (CT) antigen in multiple myeloma (MM) (Blood2003; 101: 955–960) and that it frequently elicited immune responses in vivo in these patients, suggesting that SPAN-Xb may be a suitable target for immunotherapy of MM. SPAN-Xb, however, was only detected in the tumor cells from 20% of MM patients. To increase the applicability of a SPAN-Xb tumor vaccine for MM, we have set out to determine if SPAN-Xb expression could be upregulated by pharmacologics in 8 MM cell lines.
Since most CT antigen expression occurs due to promoter demethylation, we first determined the ability of a hypomethylating agent, 5-azacytidine, to upregulate SPAN-Xb expression. Prior to exposure to 5-azacytidine, SPAN-Xb transcripts were only detected by RT-PCR in ARP-1 and RPMI 8226 cells. Following incubation in 5-azacytidine (2 uM) for 96 hours, the levels of SPAN-Xb transcripts were upregulated, as determined by RT-PCR and real time PCR, in all MM cell lines (ARK-B, ARP-1, H929, IM9, MM1-R, RPMI 8226 and U266) except KMS-11 (Table 1). This was associated with a corresponding increase in the level of SPAN-Xb protein expression, as shown by immunocytochemistry using a murine monoclonal antibody directed at SPAN-Xb.
We next determined if SPAN-Xb gene expression could be increased by cytokine pre-incubation of these cells. IL-2 (100 IU/ml) and IL-4 (1000 IU/ml) did not affect SPAN-Xb gene expression. On their own, IL-7 (0.1 ng/ml) and GM-CSF (10 U/ml) did not affect SPAN-Xb gene expression significantly. However, when combined, especially with 5-azacytidine, a summative effect of these pharmacologics on SPAN-Xb gene expression, as determined by real time PCR, was observed in IM9 cells (Table 2). Our results indicate the potential to upregulate SPAN-Xb expression in MM for therapeutic purpose using 5-azacytidine, GM-CSF and IL-7 and provide insight into the possible mechanisms of regulation of SPAN-Xb gene expression.
Table 1
Cells pre-5-azacytidine post-5-azacytidine SPAN-Xb mRNA copy numbers before and after 5-azacytidine exposure ARK-B 0 456 ARP-1 260 1376 H929 0 116 IM9 0 532 KMS-11 0 0 MM1-R 0 428 RPMI 8226 272 1348 U266 0 984
Table 2
Pharmacologics SPAN-Xb mRNA SPAN-Xb mRNA copy numbers after exposure to combination pharmacologics 5-azacytidine 532 IL-7 4 GM-CSF 9 IL-7 + GM-CSF 36 5-azacytidine + IL-7 1088 5-azacytidine + GM-CSF 952 5-azacytidine + IL-7 + GM-CSF 1632 medium only 0
Phase 2 Study of Three Doses of Single Agent Bortezomib in Patients with Fludarabine-Refractory B-Cell CLL.
Authors: Faderl Stefan , Rai Kanti R. , Gribben John , Flinn Ian , Byrd John C. , McConkey David , Schenkein David , Esseltine Dixie , Browning Mary L. , Keating Michael J. 
Published: April 2020

Abstract
Bortezomib (formerly PS-341, VELCADE®) is a selective inhibitor of the ubiquitin-proteasome pathway, which plays an important role in degrading regulatory proteins that control cell cycle functions, neoplastic growth and tumor spread. It has been approved by the FDA for the treatment of relapsed and refractory myeloma patients (pts). Phase I and II trials have suggested activity in B-cell indolent and aggressive lymphomas including mantle cell lymphoma. Preclinical studies have also shown that B-CLL cells undergo apoptotic cell death following inhibition of the proteasome pathway. We therefore conducted a multicenter phase II study to assess the safety and efficacy of 3 doses of bortezomib in pts with B cell CLL who were refractory to or intolerant to fludarabine. Pts with an ANC ≥ 1 x 109/L and platelets ≥ 30 x 109/L were eligible. Bortezomib was given as i.v. push on days 1, 4, 8, and 11 of a 21 day course for a maximum of 9 courses. Pts were initially randomized to 1.0 (n=5) versus 1.3mg/m2/dose (n=9) and later, after review of preliminary data, to 1.3 versus 1.5mg/m2/dose (n=8). Twenty-two pts were enrolled. Median age 62 yrs (range 46-83). Median number of prior therapies 4 (2–11). Median number of yrs from diagnosis to therapy 5.5 (0.8–14.2). b2-microglobulin &gt; 5.5 mg/L in 10 pts (45%). Fifteen pts (68%) had ≥ Rai stage 3. Of 19 evaluable pts no responses according to NCIWG criteria were demonstrated [stable disease 8 pts (42%), progressive disease 11 pts (58%)] based on investigator assessment. When analyzed by specific sites of disease, five pts (23%) experienced ≥ 50% decrease in ALC and 6 pts (27%) showed ≥ 50% decrease in lymphadenopathy. Bortezomib was well tolerated throughout all 3 dosing groups. Most common toxicities were ≤ grade 2 and included nausea (n=9;42%), headache (6;27%), diarrhea (5;23%), vomiting (5;23%), dyspnea (5;23%), anorexia (5;23%), and peripheral neuropathy (3;14%/ grade 3 in 1 pt). Pharmacodynamic evaluation of 20S proteasome inhibition at 1 hour postdose was similar to that reported for these doses in other studies, and was highest in the 1.5mg/m2 dosing group. We conclude that bortezomib as single agent is not effective in pts with fludarabine-refractory B-cell CLL at these doses. However, in view of the evidence of clinical activity, further exploration of bortezomib in combination with other agents might be the appropriate next step to define the optimal role of this new agent in the management of advanced CLL.