bluuebunny/crossref_metadata_2025 huggingface dataset:
Antitumor Activity of Cda-2, An Extract from Unine, in a High-Risk Myelodysplastic Syndrome Cells in Vitro
Authors: Qiu Lin , Wang Xiao-dan , Han Bing-hong , Zhan Zhao-min , Bo-long Zhang , Ma Jun
Published: April 2020

DNA methyltransferase inhibitors (DNMTI), including 5-azacytidine and 5-aza-2′- deoxycytidine, are a new class of epigenetic drug, which exhibit higher response rates in myelodysplastic syndrome (MDS) patients. Cell differentiation agent (CDA-2) is a kind of urine extracts, which contains several DNMTIs. A phase IV clinical trials for MDS showed total response rate is 69.22%. In the present study, we investigated the mechanism of CDA-2 on MDS using high-risk MDS cell line namely MuTz-1. MTT assay results showed that CDA-2 significantly inhibit the cell growth at a dose and time-dependent manner. Flow cytometer anlyasis showed that this growth inhibition was remarkblely associated with cycle arrest in G1-phase, but not associated with apoptosis. In addition, CDA-2 increased the expression of CD11b/CD14, a pair markers representing cell differentiation. we found the spectrum of hypermethylated tumor suppressor genes (TIMP3, CDKN2B, CHFR, CD44, RASSF1, TP73, IGSF4, CDH13 and DAPK) in MuTz-1 cells by Methylation-Specific Multiplex ligation-dependent Probe amplification (MS-MLPA), but the hypermethylation of these genes were remarkable decreased, as well as the expressions of DNA methyltransferase genes DNMT1 and DNMT3B at mRNA and protein level were downregulated in the treatment for 3 days with CDA-2. Also, we used CDA-2 for treatment of three MDS patients, whose several tumor suppressor genes are hypermethylated. After tour weeks of treatment, all the hypermethylation genes were undetected, part of this result was verified by methylation specific PCR (MSP) and bisulphite sequencing. In conclusion, our results demonstrated that CDA-2 may be an effective agent targeting hepermethylated tumor suppressor genes on MDS.
Incidence and Clinical Significance of Scoring System in Myelodysplastic Syndromes Including Hematological and Cytogenetic Profiles from a Single Institution
Authors: Kim Myungshin , Lim Jihyang , Kim Yonggoo , Han Kyungja , Kim Yoo-Jin , Cho Seok-Goo , Min Woo-Sung
Published: April 2020

Background: Mydelodysplastic syndromes (MDS) are clonal stem cell disorders characterized by hypercellular marrow with ineffective hematopoiesis determining peripheral cytopenia. Recently, the WHO developed a new classification that identifies the following subtypes: RA, RARS, 5q- syndrome (MDS 5q-), RCMD, RCMD with ringed sideroblasts (RCMDS), RAEB-1, RAEB-2 and MDS unclassifiable (MDS-U). The goals of our study were to establish the incidence of chromosome abnormalities in MDS patients, to correlate chromosomal defects with WHO subtype and peculiar clinical-biological findings.

Patients and Methods: We studied 242 consecutive MDS patients who were admitted in the hematology department of St. Mary’s hospital from 2000 to 2007. The diagnosis was made according to the World Health Organization classification system. The chromosome analysis of the BM cells at diagnosis was performed by the G-staining method with trypsin and karyotyped according to the ISCN 2005.

Results: The median age of patients was 50 (2–84). WHO classification was as follows: RA (n=21, 8.7%), RARS (n=7, 2.9%), RCMD (n=62, 25.6%), RCMDS (n=6, 2.5%), RAEB-1 (n=63, 26.0%), RAEB-2 (n=74, 30.6%), MDSU (n=7, 2.9%), MDS 5q- (n=2, 0.8%). Chromosomal abnormalities were found in 51.2% of patients. The cases with good, intermediate and poor cytogenetic risk features were observed in 55.0%, 29.3%, and 15.7%. Trisomy 8 was most common abnormality in all cases. 1q+, 5q-, 20q- were followed. By International Prognostic Scoring System (IPSS) 13 (5.4%) patients were classified in low group; 141 (58.3%) intermediate-1, 60 (24.8%) intermediate-2, and 28 (11.6%) high were classified. By WHO classification-based prognostic scoring system (WPSS), 14 (5.8%) patients were classified in very low group; 25 (10.3%) low, 57 (23.6%) intermediate, 110 (45.5%) high, and 36 (14.9%) very high were classified. Thirty-six patients (14.9%) progressed into AML. Both IPSS and WPSS was shown to predict survival and leukemia progression, even in a short-term prospective study (P<0.001).

Conclusions: It is important to diagnose MDS accurately on the base of cytopenia, bone marrow morphology and clonal chromosomal abnormalities. IPSS and WPSS provide good prediction of survival and risk of leukemic evolution in MDS patients. The evaluation of clinical, hematological and cytogenetic analysis in more cases for long time follow-up will be helpful to standardize diagnosis of MDS.
Intrinsic Drug Resistance in Multiple Myeloma to Doxorubicin Is Mediated by Soluble Factors
Authors: Sullivan Daniel , Dawson Jana L. , Ciaravino Elizabeth J. , Turner Joel G.
Published: April 2020

Drug resistance continues to be a major obstacle in the treatment of multiple myeloma. Data have shown that the bone marrow microenvironment plays an important role in drug resistance. Whether it is mediated through the physical interaction of stromal and tumor cells or by soluble factors, this communication is essential for survival by both normal and tumor cells. In the current study we use a transwell system to allow passage of soluble factors from high density (drug resistant) to low density (drug sensitive) human myeloma 8226 cells. In our model, 8226 cells translocate topoisomerase IIa (topo IIa) from the nucleus to the cytoplasm when at high density (3 × 106 cells/ml) for 16 hours, rendering the cells 38-fold resistant to doxorubicin (DOX), a topo II poison, relative to low density cells (see column 1 in graph A & B below). When low density (2 × 105 cells/ml) cells are separated from the high density cells by a 3.0 micron porous membrane for 16 hrs, we observed a 14-fold increase in resistance in the sensitive cells after a 1 hr exposure to DOX as compared to cells exposed to low density cells (see column 1 in graph C & D). Moreover, this increased resistance was less pronounced (2.5 fold) with topotecan (TPT), a topo I inhibitor, and no effect was seen when cis-platinum (CIS PLAT), a non-topo drug, was used in this system (see columns 2 & 3 in graph C & D), suggesting that topo IIa may be the specific target. This mechanism also appears to be time-driven, as the resistance increases over time in the sensitive cells during the course of the 16 hr exposure. In addition, drug resistance to DOX was observed when the sensitive cells were exposed to only “conditioned” media, but no effect was seen when exposed to high density cells in fresh media, suggesting that soluble factors are released from drug resistant cells into the microenvironment causing drug sensitive cells to acquire an increased ability to evade programmed cell death.

Figure Figure
Molecular Profiling of Extramedullary and Medullary Plasmacytomas
Authors: Motyckova Gabriela , Rahemtullah Aliyah , Vaghela Nileshwari , Vallet Sonia , Ackerman Adam , Mukherjee Siddhartha , Pozzi Samantha , Santo Loredana , Cirstea Diana , Toomey Christiana , Hochberg Ephraim P. , Cin Paola Dal , Rodig Scott , Hideshima Teru , Anderson Kenneth C , Munshi Nikhil , Raje Noopur
Published: April 2020

Plasmacytomas are rare plasma cells disorders and are potentially cured with local approaches such as radiation treatment. Extramedullary plasmacytomas usually progress slowly, can recur locally, and develop into multiple myeloma (MM) less frequently (8–36%) compared to solitary medullary plasmacytomas (60%). Biologically these cells are independent of bone marrow microenvironment for their growth and survival and the understanding of the differences in their underlying molecular characteristic can provide crucial insights into the pathogenesis of progression to MM. To investigate the differences between extramedullary and medullary plasmacytomas, we sought to molecularly profile these tumors by tissue microarrays, gene expression, microRNA, and FISH. We identified 78 patients from our data base with a pathological diagnosis of plasmacytoma. Of the 78 patients, 11 patients presented with extramedullary plasmacytomas, and 50 had medullary plasmacytomas. Seventeen patients who had been previously identified as having plasmacytomas were later confirmed to have MM. Among the patients with extramedullary plasmacytomas (n=11), 1 patient presented with multiple lesions. Three of 11 (27%) patients progressed to develop MM at a median of 12 months. Fifty patients presented with medullary plasmacytomas, of which 23 had solitary lesions and 27 (54%) progressed to MM at a median of 20.5 months. There was a male preponderance (76% vs 34%) and the median age at diagnosis was 59.6 years (range 30.4–84.3). Tissue microarrays were performed on 52 patients (7: extramedullary, 29: medullary, 16: MM) in whom paraffin-embedded tissue was available. Samples were stained for CD138, CD27, CD56, cyclin D1, Bcl-2, Ki-67 and light chain restriction by immunohistochemistry and/or in situ hybridization. Both paraffin and fresh frozen tissue was available for 12 medullary, 3 extramedullary, and 2 MM samples at diagnosis for comparison. FISH for del 13q, t(4;14), t(11;14), and t(14;16) was performed on these samples. Gene and microRNA profiling was also performed. The differential profile of gene and microRNA expression between the two subtypes of plasmacytomas and MM will be presented. Differential expression patterns of factors involved in proliferation, survival, adhesion, and stroma-cancer cell interactions may help explain plasmacytoma biology and identify factors responsible for progression to multiple myeloma. These insights may help identify new therapeutic approaches and targets in the treatment of plasma cell disorders.
Bortezomib-Thalidomide-Dexamethasone as Primary Induction Therapy for Newly Diagnosed Multiple Myeloma Significantly Decreases Bone Resorption While Sparing Bone Formation as Compared to Thalidomide-Dexamethasone
Authors: Tosi Patrizia , Zamagni Elena , Tacchetti Paola , Perrone Giulia , Ceccolini Michela , Brioli Anna Maria , Pallotti Maria Caterina , Pantani Lucia , Petrucci Alessandro , Baccarani Michele , Cavo Michele
Published: April 2020

Bone disease occurs in approximately 80% of patients with newly diagnosed multiple myeloma (MM) and is caused by the interaction of the neoplastic clone with bone marrow microenvironment, ultimately resulting in an altered balance between bone resorption and bone formation. It has been previously reported that therapies aimed at eradicating the myeloma clone could contribute to decrease bone resorption, even though bone formation remains impaired even in responding patients, due to the use of high-dose steroids. It has been recently demonstrated, both in vitro and in animal models, that Bortezomib improves bone formation by stimulating osteoblasts. In order to test whether this activity was retained also in vivo, we evaluated markers of bone resorption (serum crosslaps) and bone formation (serum osteocalcin-OC and bone alkaline phosphatase - BAP) in a series of patients who were enrolled in the “Bologna 2005” phase III clinical trial at our Center. By study design, after registration patients were randomized to receive three 21-days courses of induction therapy with either VTD (Bortezomib, 1.3 mg/sqm on d 1, 4, 8, and 11, plus Dexamethasone, 40 mg on each day of and after Bortezomib administration plus Thalidomide 200 mg/d from d 1 to 63.) or TD (Thalidomide as in VTD and Dexamethasone 40 mg/d on d 1–4 and 9–12 of every 21-d cycle), prior to stem cell collection and double autologous stem cell transplantation. As of January 2008, 27 patients (19 male and 8 female, median age = 57.5 yrs) entered the sub-study; of these, 15 and 12 patients were randomized in the VTD and TD arm, respectively. At diagnosis, both groups of patients showed a marked increase in serum crosslaps as compared to upper baseline limit (7321±1445pmol/L in the VTD arm and 11140±2576pmol/L in the TD arm) while both OC and BAP were reduced as compared to lower baseline limits. After completion of the induction therapy, serum crosslaps were significantly decreased in both treatment groups (2747±319pmol/L in VTD arm, p=0.007; 3686±1084pmol/L in the TD arm, p=0.0015). In the TD group a significant further reduction in bone formation markers was also observed (42% reduction in serum OC and 30% in BAP, p=0.03 and 0.04 as compared to pre-treatment values); on the contrary, in the VTD arm both OC and BAP were not significantly decreased as compared to baseline values (15% and 11% for OC and BAP, respectively). These data suggest that incorporation of Bortezomib into induction therapy counteracts the inhibitory effects of high-dose steroids on osteoblastogenesis, thus sparing bone formation.
Association of Elevated Circulating Proteasome Levels at Diagnosis with Poor Outcome in Multiple Myeloma
Authors: Liebisch Peter , Sixt Stephan , Bachmann Hagen S , Frey Ulrich H , Peters Juergen , Doehner Hartmut , Duehrsen Ulrich , Nückel Holger
Published: April 2020

Background: The proteasome is a proteolytic complex for intracellular degradation of ubiquitinated proteins which are involved in cell cycle regulation and apoptosis. A constitutively increased proteasome activity has been found in myeloma cells. Recent studies have shown that inhibition of the ubiquitin-proteasome system can be successfully used as a targeted therapy in multiple myeloma (MM). Recent data suggest a significant correlation between circulating proteasome levels (CPL) and outcome in patients with MM. Therefore, we investigated CPL in 110 patients in order to assess the role of CPL in MM.

Experimental design: CPL were measured in serum samples from healthy controls (N=10) as well as from patients with monoclonal gammopathies of undetermined significance (N=27), indolent MM (N=15) and symptomatic MM (N=68) using enzyme-linked immunoabsorbent assay (ELISA) techniques detecting circulating 20S proteasome components. All serum samples were collected at the University Hospital in Ulm at time of diagnosis.

Results: The median CPL were 123.5 ng/mL (range, 95–185 ng/mL) in healthy controls, 180 ng/mL (range, 100–485 ng/mL) in patients with MGUS (N=27) or indolent MM (N=15), and 227.5 ng/mL (range, 100–985 ng/mL) in patients with symptomatic MM (N=70). The CPL of patients with symptomatic MM were significantly elevated compared with healthy donors (p=0.0017) and to persons with asymptomatic gammopathies (p=0.046). While CPL were also significantly higher in the MGUS/indolent MM cohort as compared to controls (p=0.03), CPL in MGUS and indolent MM were comparable. Using ROC analysis in the symptomatic MM cohort patients with CPL >150ng/mL (N=50) had a significantly shorter progression-free survival (PFS) time than patients (N=18) with CPL<150 ng/mL levels (median PFS: 40 versus 57 months, log rank test p=0.026). Of note, all six patients who died in the observation interval had CPL >150ng/mL.

Conclusions: Here we demonstrate that increased CPL at diagnosis correlates with poor outcome in symptomatic MM patients. Evaluation of the prognostic significance of CPL in a larger cohort of uniformly treated patients with symptomatic MM is currently under way. This data will be presented at the meeting.
The Evaluation of Light Chains USING ELISA Method
Authors: Vavrova Jaroslava , Friedecky Bedrich , Tichy Milos , Holeckova Magdalena , Maisnar Vladimir , Hajek Roman
Published: April 2020

Introduction: Free light chain assay (FLC) has moved into clinical practice based on the building evidence of its utility in multiple myeloma (MM) and other monoclonal gammopathies. The purpose of the study is to verify the effectiveness of the basic parameters relevant to serum free light chain analysis using ELISA method and to compare it with the well-established FreeLite assay.

Methodology and sample collection: ELISA method was tested using diagnostic kit developed by Biovendor (Czech Republic). Immunoturbidimetric measurement using the FreeLite kit (The Binding Site), adapted on the Modular P (Roche), and was used as a reference method. Regression analysis, developed by Passing and Bablok, expressed by equation Y = −9,84 + 1,53 X allowed to compare the two methods. In total, the results of serum FLC determination in 40 donors and in 202 patients with monoclonal gammopathies were compared.

Results: Reference intervals were tentatively determined based on an analysis of male and female blood donor samples. The ratio kappa/lambda was from 0.24 to 1.17 by use of ELISA method and from 0.53 to 1.92 by reference immunoturbidimetric method. Intermediate measurement precision was obtained as follows: (intermediate) kappa using ELISA 6,6–11,0 %, using immunoturbidimetric method 5,6–6,9 %; lambda using ELISA 7,3–8,6 %, using immunoturbidimetric method 6,1–9,2 %. Regression coefficient was R = 0, 73. We were observing the influence of sample dilution on the results and we confirmed the necessity to verify the degree of dilution in each pathologically changed sample. The main factors of a successful free light chain determination are: validity of the reference intervals, difference in diagnostic classification when using different methods. Considering the necessary dilution, every patient sample requires a special and individual attention. In many patients, an applicable result can be obtained only having repeated the measurements at different dilution levels; therefore, it is not possible to predict the number of examinations one kit can serve to.

Conclusion: The determination of serum free light chains cannot be considered as a routine procedure for the time being. This analysis requires careful sample preparation and high professional level of laboratory personal. Both methods used to determine serum FLC are suitable for laboratory practice; the comparability of their predictive value in clinical practice requires a further testing.
Assessment of Thalidomide Therapy Effect on Platelet Activation and Selected Blood Coagulation Parameters in Multiple Myeloma Patients
Authors: Robak Marta , Trelinski Jacek , Chojnowski Krzysztof
Published: April 2020

Background: Patients with multiple myeloma are at relatively high risk of developing thromboembolic events (TEE). These life-threatening complications may arise from hypercoagulability associated with malignancy and/or may be connected with anticancer therapy. The risk of developing TEE appears to be particularly high during treatment with thalidomide alone or combined with chemotherapy and/or high-dose dexamethasone. The pathogenesis of thalidomide-related thrombosis in myeloma patients remains unexplained. Some authors suggest that platelet activation can contribute to development of this complication in multiple myeloma patients on thalidomide therapy but until now it has not been a subject of investigation.

Patients and methods: The study was performed in 20 patients with multiple myeloma. The tests were done at diagnosis and after one month of thalidomide therapy at a dose of 100–200 mg/24h. All patients had normal renal function and did not take drugs affecting platelet function. The control group consisted of 15 healthy subjects of similar age. In each patient closure time with ADP/Collagen and Epinephrine/Collagen cartridges by PFA-100 method was assessed. Platelet expression of membrane activation marker P-selectin (CD62p) on resting platelets and after stimulation with ADP/Collagen and Epinephrine/Collagen was analyzed by flow cytometry. Additionally, activity of factor VII, factor VIII and von Willebrand factor (vWF), concentration of fibrinogen and D-dimer, and platelet count were evaluated.

Results: The mean PFA-100 closure time was significantly shortened with ADP/Collagen (87.2 ±17.1 s vs 100.4 ± 19.3 s, p=0.008) and Epinephrine/Collagen cartridges (118.5 ± 20.3 s vs 132.2 ± 27.9 s, p=0.04) after one month of therapy in comparison to baseline. The median CD62p percentage increased markedly after treatment-on resting platelets 5.1 (0.76–22.2) vs 3.6 (0.1–21.5) p=0.03. and after stimulation with Epinephrine/Collagen 16.6 (2.3–57.3) vs 11.1 (1.4–19.5) p=0.03. The observed increased P-selectin expression after ADP/Collagen stimulation 26.3 (8.5–42.8) vs 19.7 (1.0–35.4) was not statistically significant. The median values of P-selectin expression at diagnosis and after thalidomide therapy were also higher than in the control group. The results of factor VIII, vWF activity, fibrinogen and D-dimer concentration did not differ markedly before and after therapy. Significantly lower mean activity of factor VII (p=0.004) and higher mean platelet count (p=0.03) after therapy were observed.

Conclusions: These results demonstrate that platelet activation is one of the pathogenetic factor of thalidomide-related thrombotic complications and can explain some observations that acetylsalicylic acid may protect against TEE during myeloma treatment with thalidomide.
An Observational Post Authorization Study on the Use of Bortezomib in Multiple Myeloma Patients in the Netherlands: Results of An Interim Analysis
Authors: Lokhorst Henk , van Der Heul Cornelis , Roerdink Henk , de Weerdt Okke
Published: April 2020

Introduction: Bortezomib (Velcade) is a first-in-class proteasome inhibitor for the treatment of patients (pts) with multiple myeloma (MM). Most data on efficacy and safety are from randomised controlled studies in selected groups of pts with MM.

Aim of the study: To evaluate safety and efficacy of bortezomib in the real-life community oncology practice setting as a second-line therapy or in subsequent lines of therapy in pts with MM.

Materials and methods: In an observational, multicentre study, the efficacy and safety of treatment with bortezomib was assessed in MM pts. An interim analysis was performed on data of pts enrolled from November 04 until November 06 (109 pts).

Results: All results are given as median. The pts age was 67 yrs, Karnofsky Performance Score was 80%, time from initial diagnosis was 3.3 years, 62% of the pts had more than 2 previous lines of treatment, 85% of the pts had disease stage III. Pts were treated with median 4 cycles (95% CI: 4.1–5.0) of bortezomib. Median time to initial response was 50 days and median time to best response category was 84 days. Of all pts 43.2% achieved a partial response or better (2.8% CR, 6.4% nCR, 3.7% VGPR, 30.3% PR); 16 pts were not evaluable for response. The duration of response (DOR) was 6.3 months, PFS was 5.8 months and overall survival was 1.2 yrs. The treatment was generally well tolerated. Most common adverse events (all grades) in >15% of the pts were, in order of decreasing frequency, thrombocytopenia, nausea, diarrhoea, fatigue, (poly)neuropathy, constipation, malaise, anaemia, pyrexia, vomiting and anorexia. For responding pts treatment discontinuation due to an AE occurred in 46.8% of the pts. For the three most common AE’s (polyneuropathy, fatigue/malaise, thrombocytopenia), resulting in discontinuation of the bortezomib treatment, the discontinuation was not preceded by a dose adjustment in almost half of the cases. In 9 pts (8.3%), Herpes zoster reactivation was reported. Twenty-two out of 109 pts were thalidomide naïve when starting bortezomib. 59.4% of these pts achieved a PR or better vs. 39% in thalidomide pre-treated pts, but thalidomide pre-treated pts had a longer time from initial diagnosis (3.5 vs 2.6 years) and had more previous treatment lines compared to thalidomide naïve pts (3 2 lines of previous therapy 97.7% vs 59.1% respectively).

Conclusion: Response rates in this study were comparable to those in other community studies and higher than reported in previous clinical studies. Overall the treatment with bortezomib was well tolerated. However, it seems dose adjustment as a strategy to continue treatment and to increase the quality of response, is still underutilized in clinical practice. Overall survival was as expected in this patient population. The thalidomide naïve group was yet too small to draw any conclusions on safety and efficacy as compared to the thalidomide pre-treated pts.
IgM Multiple Myeloma: A Rare Subtype Highly Sensitive to Lenalidomide
Authors: Moreno-Aspitia Alvaro , Charles Antony , Patel Tejal , Bueno Celine , Zubair Abba , Menke David
Published: April 2020

Background: IgM multiple myeloma (MM) are very rare plasmaproliferative disorders representing 0.5–1.2% of all cases of MM and < 0.2% of all IgM monoclonal gammopathies. Clinical criterion are not always helpful in differentiating IgM MM from Waldenstrom macroglobulinemia. However, the presence of lytic bone lesions, absence of lymphadenopathy and/or hepatosplenomegaly, presence of translocation of the immunoglobulin heavy chain locus at 14q32 [t(11;14), t(14;16), t(4;14)], and strong expression of CD138 by the plasma cells are useful in the diagnosis of IgM MM. It has been our experience and of others that these cases have an aggressive behavior at presentation, shorter survival than IgG and IgA MM and poor response to therapy for lymphoplasmacytoid lymphomas. We present here 2 cases of IgM MM with a dramatic response to Lenalidomide and low dose dexamethasone (Rev/Dex)

Results: Baseline patient characteristics at time of diagnosis of IgM MM and therapy outcome are presented in the following 2 tables:

Table 1.

Case 1 2 Age and sex 72 (F) 73 (F) Serum M-spike (g/dL) 5.3 6.2 Urine M-spike (mg/dl/24 hrs) 72 412 Serum IgM (mg/dL) 8,590 11,000 BM plasma cells percentage 90 20 Plasma cell immunophenotyping CD138+++, partial CD20, CD56− CD138+++, partial CD20, CD56− Cytogenetics (Standard and/or FISH) Standard: normal FISH: not done on initial biopsy. On follow up there were insufficient number of plasma cells to perform test Standard: of 20 metaphases, 6 had a complex hypotetraploid karyotype with relative loss of 13q, 14, 15, 16, 20, and 22, and numerous unbalanced rearrangements. FISH: a plasma cell clone with monosomy 13 and IGH/c-MAF fusion, t(14;16). In addition, approximately 60% of plasma cells had a tetraploid clone with the same anomalies as well as relative loss of p53 Bone lesions Multiple non-traumatic spinal fractures and of stenum Several lytic lesions of long bones Renal insufficiency No No Anemia (Hbg g/dL) Yes (8.7) Yes (8.1) Hypercalcemia (Ca mg/dL) Yes (12.5) Yes (11.4) Beta 2 microglobulin (mg/dL) 5.79 8.51 Serum viscosity (cpoise) 5.9 4.8

Table 2. Best Response to therapy

Case Therapy Best Response Comments 1 Rituxan, then Fludarabine based therapy Transient response Rapid progression after partial and transient response to each therapy 1 Lenalidomide + LD-Dex sCR after cycle #6. Currently on CR 18 months later IgM declined from 8,590 to 43 mg/dL after 4 cycles of Rev/Dex. 2 Lenalidomide + LD-Dex VGPR after cycle #2 IgM declined from 11,000 to 463 mg/dL after cycle 3. Complete disappearance of M-spike in serum; BM to be done after cycle #4

Conclusions: This is the first report that we are aware of a rapid and dramatic response to lenalidomide and low dose dexamethasone in these rare cases of IgM MM with poor response to NHL-type treatment. Lenalidomide-based therapy might abrogate poor prognosis cytogenetics in this unusual subtype of MM (case #2), however, follow up for this patient is still very short.
The Role of An Early Interim 18 F-FDG PET/CT for Prediction of Outcomes of Patients with Peripheral T Cell Lymphoma
Authors: Choi Yoon Hee , Kang Byung Woog , Hong Hyun Soo , Kim Chul , Jang Geundoo , Lee Sung Sook , Ryu Jin Sook , Huh Jooryung , Lee Dae Ho , Kim Sang We , Lee Jung Shin , Suh Cheolwon
Published: April 2020

Background Recent studies have demonstrated that positron emission tomography (PET) performed after one to four cycles of multiagent chemotherapy predicts therapeutic outcome in lymphomas which are mainly diffuse large B cell lymphoma and Hodgkin’s lymphoma. However, interim 2-[18F] fluoro-2-deoxy-D-glucose (FDG) PET/CT has not yet been discussed in peripheral T cell lymphoma. In this study, we investigated the role of an interim 18 F-FDG PET/CT for the prediction of the progression free survival (PFS) and overall survival (OS) in peripheral T cell lymphoma.

Patients and methods We retrospectively analyzed the medical records of thirty-three patients with newly diagnosed, initially FDG-avid peripheral T cell lymphoma between 2004 and 2007 (median age of 44 years, 18 peripheral T cell lymphoma, unspecified, 9 anaplastic large cell lymphoma, 6 angioimmunoblastic T cell lymphoma) who underwent 18F-FDG PET/CT after one to four cycles of chemotherapy (interim scan). The maximal response to initial therapy was evaluated using conventional methods. Interim FDG-PET/CT results were correlated to the PFS and OS using Kaplan-Meier survival analysis. Cox regression analyses were performed for independence of prognostic factors.

Results The median follow up was 23 months (range, 2–52 months). In the initial therapy, 25 patients achieved CR, 5 patients showed progress and 3 patients died. Interim FDG-PET/CT was negative in 17 patients, positive in 9, and 7 patients showed minimal residual uptake (MRU). The CR rate of initial therapy was 100%, 85% (6/7) and 22% (2/9) in the FDG negative, MRU and positive group, respectively. The estimated 2 year PFS was 88.2% for the negative group, 38.3% for the MRU group, and 0% (median survival of 5.6 months) for the positive group. The log rank test showed strong associations between interim FDG-PET/CT results and PFS (P =0.001) and OS (P <0.001). Also, univariate and multivariate analyses showed that the interim FDG-PET/CT was the only independent factor to predict outcomes among known prognostic factors such as age, elevated lactate dehydrogenase, the number of extranodal disease, stage, performance status (p=0.007).

Conclusion Interim FDG PET/CT is an excellent and independent predictor of outcomes of patients with peripheral T cell lymphoma and is suggested to be a useful tool to modify the therapy.
Prognostic Value of P-Glycoprotein (Pgp) in Non-Hodgkin’s Lymphoma
Authors: Pilleux Lilian M , Carrasco Cristian S , Pisón Claudia V , Valencia Yenny O , Lopez Mauricio G , Benavides Tatiana , Calderón Susana E
Published: April 2020

Multidrug resistance (MDR) is one of the greatests obstacles of cancer therapy. One of the most studied mechanisms of resistance is mediated by Pgp encoded by gene MDR-1. Studies in patients with Human Immunodeficiency Virus associated non-Hodgkin’s lymphoma (NHL) show that those MDR positive achieve lower rate of complete remission than those MDR negative. Reports in refractory or relapsed NHL have failed to reproduce this finding. In an attempt to confirm the relationship between Pgp and poor outcome in NHL we analyzed the expression of Pgp in patients treated in our cancer reference hospital.

Patients and Methods: We retrospectively analyzed a cohort of patients 15 years or older diagnosed with NHL in 1998–2002 quinquennium in Valdivia Public Hospital (VPH) treated in accordance with the protocols of the National Adults’ Antineoplastic Drugs Program (NAADP) with an emphasis on clinical response and histological features. An immunohistochemical staining (IH) with Pgp Clone C494 considering negative that one without positivity or <5% of the cells, + positive: 5–25%, + + positivity 25–50% and + + + positivity 50–100%. We performed a descriptive analysis and Kaplan-Meier actuarial survival analysis using log-rank and Wilcoxon statistical tests for comparing curves. It was considered statistically significant value of p <0.05.

Results: During the study period 96 patients were admitted to NAADP at VPH corresponding 82 to newly diagnosed cases confirmed histologically, 14 were excluded (6 relapses, 4 histology not NHL, 2 without biopsy, 2 not evaluable). At the time of analysis there were 49 cases of with Pgp IH. 63% of the cases were positive and 37% negative. Among the positive cases Pgp intensity was: + in 34.5%, + + in17.2% and + + + in 48.3%. When analizing survival curves according to Pgp positivity at 2 and 5 years, respectively, we observed that:

The overall survival (OS) showed a trend towards increased survival in the Pgp negative group, but without significant difference (log-rank p = 0.07; Wilcoxon p = 0.05). Disease-free survival (DFS) showed no differences (log-rank p = 0.49; Wilcoxon p = 0.65).

The OS according to intensity of Pgp positivity again showed no differences (log-rank p = 0.07; Wilcoxon p = 0.11). When analizing OS in Pgp positive cases according to localized (I and II) or advanced (III and IV) stages we observed it was higher in the first group (log-rank p = 0.01; Wilcoxon p = 0.03).

Conclusions: In our study the expression of Pgp or its intensity at the time of diagnosis in patients with NHL did not have prognostic value. We have to consider the limited number of cases analized and the low percentage of localized stages in it. Additionally histological subtypes Pgp negative cases were mostly aggressive. Further studies are needed to discard Pgp expression as another prognostic factor in NHL. (DID S-200725 Proyect)
The Risk of Venous Thrombosis among Women with Thrombophilia and Oral Contraceptive Use: A Meta-Analysis
Authors: Dayan Natalie , Tagalakis Vicky , Holcroft Christina
Published: April 2020

Introduction: Several small studies have reported an increased risk of venous thrombosis (VT) with thrombophilic disorders that is compounded by the use of oral contraceptives (OCP). The objective of this meta-analysis was to summarize the risk of VT among women with thrombophilia and OCP use, and to assess the interaction between OCP use and thrombophilia.

Methods: Data sources used included MEDLINE, EMBASE, CINAHL, and COCHRANE REVIEW databases (1950–2008), and reference lists of retrieved articles. We selected studies that assessed among reproductive-aged women with and without VT the prevalence of OCP use and thrombophilic disorders, including factor V Leiden (FVL), prothrombin (PT) G20210A, elevated procoagulant factor VIII, hyperhomocysteinemia, lupus anticoagulant, and deficiencies of proteins C, S and antithrombin III. VT was defined as objectively confirmed deep vein thrombosis, pulmonary embolism or cerebral vein thrombosis. Fifteen studies were included. Two independent reviewers extracted data on study characteristics, quality, risk factors, and outcomes. Odds ratios (ORs) were calculated for each study and pooled using the random effects model.

Results: There was an increased risk of VT in women with OCP use (pooled OR 3.0, 95% confidence interval (CI) 2.0–4.5), women with any thrombophilic disorder (pooled OR 4.8, 95% CI 3.6–6.5), women with FVL (pooled OR 4.7, 95% CI 3.1–7.5), and women with PT G20210A (pooled OR 3.3, 95% CI 0.8, 13.1). OCP users with FVL had a 18-fold increased risk of VT compared to healthy OCP non-users (pooled OR 17.8, 95% CI 5.6–56.8) and OCP-using women with PT G20210A had a 12-fold increased risk (pooled OR 12.4, 96% CI 0.9– 166) compared to healthy OCP non-users. The association of OCP use and VT was of similar magnitude among women with FVL (pooled OR 3.2, 95% CI 1.3–7.7) and women without FVL (pooled OR 3.9, 95% CI 1.8 to 8.6). Significant heterogeneity (I2 >80%) was present among models for OCP use, PT G20210A, and OCP use among FVL-negative women.

Conclusions: We have shown that OCP use, thrombophilia, and in particular FVL and PT G20210A are associated with an elevated risk of VT of similar magnitude. However, the combination of OCP use and thrombophilia further amplifies the risk of VT compared to when either of the risk factors are considered alone. Though heterogeneity was observed, our study suggests, and confirms previous reports of an interaction between OCP use and thrombophilia.
Prevalence of Thrombophilia in Patients with Retinal Vascular Occlusion
Authors: De Sancho Maria Teresa , Powell-Boone Karen , Carlson Karen
Published: April 2020

Background: Retinal vascular occlusion (RVO), both arterial (RAO) and venous (RVO) is an important cause of visual loss. Inherited and acquired thrombophilia are considered important risk factors in the development of RVO. Additionally, cardiovascular risk factors such as hypertension, diabetes, hyperlipidemia, smoking and use of female hormones may play a role.

Objective: To determine the prevalence of thrombophilia in patients with RVO.

Methods: We reviewed the medical records of all patients with documented RVO who were referred to our hematology clinic at a tertiary care center between January 2001 and June 2008 for thrombophilia evaluation. Thrombophilia evaluation included testing for factor V Leiden, prothrombin gene mutation G20210A, deficiencies in antithrombin, protein C and S, presence of antiphospholipid antibodies (aPLas), hyperhomocysteinemia and increased levels of lipoprotein (a), factor VIII, IX and XI. Serum protein electrophoresis and immunofixation were also performed. Environmental risk factors analyzed included use of female hormones, pregnancy and puerperium, smoking, hypertension, hyperlipidemia and diabetes.

Results: A total of 24 patients were identified; 10 were males and 14 females. The mean age was 62 years (range, 31 to 86 years). Seven patients had RAO and 17 RVO. Of the 24 patients, 19 (79%) had thrombophilia. Of these, 10 had only one thrombophilia and 9 had ≥2 thrombophilia. Of the 10 patients with a single thrombophilia, 3 patients had aPLas, 3 had increased lipoprotein (a) 2 had factor V Leiden, 1 hyperhomocysteinemia, and 1 had increased factor VIII level. Of the nine patients with combined thrombophilia: 3 had hyperhomocysteinemia and increase factor VIII level, 2 prothrombin gene mutation and positive lupus anticoagulant, 1 positive aPLas, hyperhomocysteinemia and a monoclonal gammopathy, 1 aPLas and increased factor VIII, 1, hyperhomocysteinemia and protein S deficiency and 1 hyperhomocysteinemia and polycythemia vera. In 5 patients, no thrombophilia was identified. Of the 24 patients, 11 had hypertension, 9 had hyperlipidemia, 3 were smokers, 3 were using female hormones and 1 was pregnant. Treatment included aspirin 81 mg (n= 9), aspirin >81 mg (n= 5), warfarin (n= 3), folic acid (n=2), aspirin and clopidogrel (n=1), clopidogrel (n=1), no treatment (n=4), and unknown (n=1).

Conclusions: Over three quarters (79%) of patients with RVO had an underlying thrombophilia with aPLas, hyperhomocysteinemia and increased factor VIII level being the most prevalent. Our findings suggest that testing for thrombophilia particularly for aPLas, homocysteine and factor VIII levels may be useful in patients with RVO
A Rare Cause of Anemia in Inflammatory Bowel Diseases: A Case Report and Review of Literature
Authors: Ahmed Waqas , Monroe Kevin , Essell James , Broun E. Randolph
Published: April 2020

Introduction: Anemia is a common problem in patients with inflammatory bowel diseases (IBD), and its etiology is usually multifactorial. It can be produced by chronic blood loss, nutritional deficiencies, and drugs such as salazopyrine; however it can also due to auto immune hemolysis, which is a rare complication of IBD. We report a case of coombs positive autoimmune hemolytic anemia associated with ulcerative colitis both diagnosed at the same presentation.

Case Report: A 32 year old man with no significant past medical history presented with complaint of dark colored urine, jaundiced skin and fatigue for 4 weeks. He also reported diarrhea mixed intermittently with blood for last few months. Physical exam was consistent with jaundice and anemia (pallor and icterus) with slightly palpable spleen. Initial lab work up showed Hb of 3.8 with normal platelet and WBC count, high reticulocytes count of 7% .LFT showed serum bilirubin of 3.6 (direct 0.4) with normal serum ALT and AST levels .Serum LDH was high (1032 U/l) while serum haptoglobin was low (0.11 mg/dl). Peripheral smear showed anisopoikilocytosis & spherocytosis. (See Figure 1) Further investigations revealed a positive direct Coombs test consistent with diagnosis of autoimmune hemolytic anemia. CT abdomen and pelvis showed mild splenomegaly & non-specific enlarged mesenteric lymph nodes. Colonoscopy revealed ulcerative pancolitis confirmed by histological findings of biopsies taken. Patient received PRBC transfusions and was started on steroids and mesalamine and was discharged on maintenance dose. His symptoms resolved in 4 weeks and Hb remained stable with no evidence of further hemolysis at 4 month follow up .Repeated CT abdomen & pelvis showed resolution of the lymphadenopathy.

Figure 1: Peripheral Blood smear showing anisopoikilocytosis & spherocytosis. Figure 1:. Peripheral Blood smear showing anisopoikilocytosis & spherocytosis.

Discussion: Autoimmune hemolytic anemia (AIHA) is a rare complication of IBD. The exact underlying pathogenesis of this association remains obscure; however it has been attributed to the production of cross reacting anti erythrocyte antibodies. In AIHA associated with IBD, corticosteroids are considered to be first line therapy and often cause remission of hemolysis along with treatment for IBD Immunomodulators and splenectomy has been used for patients with refractory AIHA. Colectomy done for fulminant colitis has also been reported to induce remission of AIHA. Further studies for long term follow up and pathogenesis of this association are warranted.
Identification of Novel Phosphorylation Sites and Kinetics in Flt3 Receptor Wt, ITD and D835Y
Authors: Razumovskaya Elena , Masson Kristina , Khan Rasheed , Bengtsson Susanne , Ronnstrand Lars
Published: April 2020

FMS-like tyrosine kinase-3 (Flt3) is a receptor tyrosine kinase, which is normally expressed in hematopoietic progenitor cells. It has been implicated as a major cause of transformation in acute myeloid leukemia (AML). There are two types of Flt3 gene mutations have been identified in AML: duplication of amino acids in the juxtamembrane region- Internal Tandem Duplication (ITD) and an activation loop point-mutation of D835 in kinase domain. These mutations cause constitutive activation or over expression of Flt3 receptor and therefore lead to alteration in signal transduction. These alterations occur in approximately 30% of AML patients. High occurrence of these mutations in the Flt3 receptor in AML patients makes it one of the most interesting therapeutic targets. In this study we have identified three novel in vivo phosphorylation sites of Flt3 receptor and further compared the activity of phosphorylation sites of Flt3 wild type, Flt3 ITD and D835Y mutations by using homemade phospho-specific antibodies directed against specific tyrosines. For this study murine hematopoietic Ba/F3 cells were stably transfected with wild-type Flt3, ITD and D835Y mutations. We have confirmed that the activation of the wild type Flt3 receptor is ligand dependent and response in a time dependent manner, but Flt3-ITD and D835Y are constitutive active and ligand independent. Phosphorylated tyrosines 589, 591, 599, 726, 768, 793, 842, and 955 of Flt3 receptor were investigated and shown to be differentially activated in wild-type versus the mutated receptor. Using this data we can further study the mechanisms of signaling pathways of the Flt3 receptor that are involved in many biological responses and understand the mechanism by which Flt3 ITD and D835Y functions in pathological conditions.
Generation of Endothelial Progenitor Cells from Amniotic Fluid Stem Cells
Authors: Cardier Jose E. , Wittig Olga , Alonso Jose , Romano Egidio
Published: April 2020

Recently it has been shown that human amniotic fluid contains stem cells of fetal origin (AFS) that express embryonic and adult stem cell markers. AFS can be expanded extensively in culture and give rise to multiple differentiated cells such those of myogenic, osteogenic, neurogenic and endothelial lineages, all of them with potential therapeutic value. In this study, we investigated the generation of endothelial progenitor cells (EPC) from AFS. For this purpose, remnant amniotic fluid was obtained from mothers undergoing amniocentesis for medical reasons at 18 weeks of pregnancy. Cells were collected by centrifugation and cultured in DMEM, supplemented with Chang medium and fetal bovine serum. Adherent cells were expanded and characterized by flow cytometry at various times using antibodies for CD34, CD45, CD133, CD117, KDR (VEGFR-2) and CD31. Cultures were also carried out in endothelial growth medium and analyzed for EPC markers (CD133, KDR, CD31). Adherent cells showed, at 8 and 17 days of culture, fibroblastoid features. Flow cytometry analysis, at 2 weeks of culture, showed cells expressing CD 133 (37%), CD 34 (1%), CD 117 (3%) and KDR (12%). Cultures carried out in endothelial differentiation medium (EDM), at 33 days of culture, showed a significant increase of cells expressing EPC markers: CD133 (40%), CD34 (3%) KDR (24%), KDR/CD31 (9%). These cells formed blood vessel-like structures on collagen-coated plates. The in vivo angiogenic capacity of these cells was demonstrated by inoculating them into collagen microspheres and implanting them subcutaneously into immunocompromised mice. Histologic examination revealed that these cells formed microvessels on implantation. Our data indicates that EPC can be generated and expanded from AFS, and they might be useful for therapeutic angiogenesis in ischemic tissues.
Changes in MRI-Estimated Cardiac and Hepatic Iron Load in β-Thalassemia Patients Treated with Deferasirox at a Single Institution
Authors: Kattamis Antonios , Moraitis Panayiotis , Lourida Athanasia , Rigatou Efthemia , Ladis Vassilios
Published: April 2020

Chronic transfusions have dramatically improved the life expectancy of patients with β-thalassemia, but leads to severe iron overload. Long-term iron chelation therapy aims both in the prevention and the reversal of iron load and iron toxicity. The available chelators seem to have different profiles of relative heart and liver iron removal. Data on cardiac efficacy of deferasirox (DFX; Exjade®) are limited. We report results on patients with thalassemia, followed in a single institution, who completed at least 12 months of treatment with deferasirox.

Methods: Fifty five patients (22 males, 33 females; mean age: 27±7.1 years, splenectomised: 5 patients, HCV seropositive: 7 patients) were included in the study. The patients were treated with deferasirox at a mean dose of 27.2±5.2 mg/kg/day (range 15–40 mg/kg/day). Sixty-one pairs of MRI assessments of cardiac and hepatic iron overload by using the T2* technique were performed at a mean time interval of 13.7±2.4 months. Cardiac function parameters, mainly ejection function (EF), were also estimated at the same time.

Results: Mean, SD and range at the time of the first MRI evaluation showed: ferritin: 2520±1584 ng/mL (371–6953 ng/mL), cardiac T2*: 25.3±10.7 ms (3.8–39.2ms), liver T2*: 4.5±4.6 ms (0.57–21.1ms) and ejection fraction: 66.9±4.8 ms (55.5–75.4%). Follow up assessments showed: ferritin: 2375±1743 ng/mL (39–7245 ng/mL), cardiac T2*: 27,1±11.7 ms (2.7–40.8ms), liver T2*: 6.2±6.2 ms (0.5–26.4ms) and ejection fraction: 66.1±4.8 ms (55.8–75.4%). Changes in liver and cardiac T2* were statistically significant (p=0.0004 and 0.008, respectively). Changes of the cardiac T2* values correlated well with the dose of deferasirox, while changes of liver T2* with ferritin changes (r=−0.284, p=0.032, r=−0. 27 and p=0.046, respectively). A statistical significant improvement (p<0.05) in cardiac and hepatic iron load was observed in the subgroup of 22 patients with severe liver iron load (defined as hepatic T2*<1.9 ms). In 18 patients with moderate/severe cardiac iron load (cardiac T2*<20ms), T2* decreased in 5, remained stable in 2 and improved in 11 patients (mean 10.7±5.5 ms vs. 12.1±7.1, p=0.078).[S1]

Conclusions: This observational study shows that treatment with deferasirox improves both hepatic and cardiac iron overload in regularly transfused β-thalassemia patients. Adjustment of the dose is required to enhance iron excretion and achieve better negative iron balance