bluuebunny/crossref_metadata_2025_split huggingface dataset:
Increased Thrombin Generation in Severe Hemophiliacs with Mild Clinical Phenotype
Authors: Santagostino Elena , Mancuso Maria Elisa , Tripodi Armando , Chantarangkul Veena , Pasta Gianluigi , Siboni Simona Maria , Mannucci Pier M. 
Published: April 2020

Abstract
Introduction: Some severe hemophiliacs (FVIII/FIX&lt;1%) exhibit a mild bleeding tendency, but the basis for this clinical heterogeneity is poorly understood. This study investigated the relationship between the values of endogenous thrombin potential (ETP) and clinical phenotype in severe hemophiliacs. The impact of FVIII/FIX gene mutations and thrombophilic polymorphisms was also evaluated. Methods: severe hemophiliacs older than 18 years without inhibitor history and treated on demand were eligible. Mild bleeders (MB) and severe bleeders (SB) were defined as follows: spontaneous bleeding episodes per year ≤2 (MB) or 25 (SB) and concentrate consumption &lt;500 (MB) or &gt;2000 (SB) IU/Kg/year. Patients who did not fit these criteria were considered as intermediate bleeders (IB). FVIII was measured by chromogenic assay and ETP was measured in platelet-rich plasma after addition of tissue factor. Results: 22MB, 22SB and 28IB were enrolled. MB had lower clinical and radiological scores when compared with both IB and SB (p&lt;0.005). MB showed an older age at first bleed compared to SB (p &lt; 0.005) and p for trend among the 3 groups was also significant (p &lt; 0.05). The prevalence of severe FVIII/FIX gene defects (null mutations) was lower and ETP values were higher in MB compared with both IB and SB (p&lt;0.05; table 1). Conclusions: our results indicate an extremely low prevalence of null mutations in severe hemophiliacs with mild bleeding diathesis. The measurement of thrombin generation in platelet-rich plasma may allow to identify this subgroup of patients, not otherwise distinguishable by conventional functional assays.
SB (#22) IB (#28) MB (#22) p Age (yr) 38 (21–76) 38 (23–62) 32 (22–73) NS Age 1st bleed (yr) 1 (0–4) 2 (0–6) 3 (1–10) &lt; 0.005 Bleeding episodes/yr 36 (25–60) 10 (3–20) 0 (0–2) &lt; 0.0005 Factor use (IU/Kg/yr) 2207 (2040–8696) 1068 (207–2400) 60 (25–487) &lt; 0.0005 Clinical score 18 (10–35) 10 (0–34) 3 (0–17) &lt; 0.005 Pettersson score 44 (14–62) 28 (0–48) 17 (3–40) &lt; 0.0005 Null mutations (%) 59 70 6 &lt; 0.005 PTG20210A (%) 0 7 5 NS FV Leiden (%) 5 7 0 NS Median ETP (nM) 414 478 850 &lt; 0.05
Clinical and Biological Features of Biclonal Gammopathies. Review of 203 Cases
Authors: Decaux Olivier , Leroy Helene , Ianotto Jean-Christophe , Ruelland Annie , Guenet Lucienne , Sebillot Martine , Jego Patrick , Grosbois Bernard 
Published: April 2020

Abstract
Introduction: Biclonal gammopathies represent about 5% of clonal gammopathies. We describe the clinical and laboratory features of biclonal gammopathies identified in a French university hospital.
Methods: Patients were selected by immunofixation registry of Biochemistry laboratory.
Results: From 1987 to 2008, 203 biclonal gammapathies were identified. Patients were 113 men and 90 women. Median age was 72.0 years (35–95). Seventy eight patients (38.3%) had IgG and IgM components, 64 (31,9%) had two IgG, 24 (11,8%) had IgG and IgA, 23 (11,3%) had two IgM, 8 (3,9%) had IgM and IgA, 5 (2,4%) had two IgA, one (0.5%) had Ig G and IgD (0,5%). Of the 406 light chains, 260 (63,8%) were kappa, 146 (36.2%) were lambda. Eighty nine patients (44.1%) had two kappa light chains, 82 (40,2%) had both kappa and lambda and 32 (15,7%) had two lambda chains. Median gammaglobulin concentration was 13.3 g/L (3–59.9). The most frequent diagnosis was biclonal gammopathy of undeterminated significance (BGUS) in 123 patients (60.6%). Others patients could be divided into 3 groups. Forty seven patients (23.1%) had lymphoproliferative diseases, including Waldenström’s macroglobulinemia (21 cases), non Hodgkin lymphoma (20), chronic lymphocytic leukaemia (6). Eighteen (8.87%) had multiple myeloma. For the last 15 patients (7.4%), biclonal gammopathy was associated with a non lymphoid hemopathy or with systemic disease. Biclonal gammopathy was identified in 12 patients already known to have a monoclonal gammopathy (7 monoclonal gammopathy of undetermined significance-MGUS, 3 myeloma and 2 Waldenström’s macroglobulinemia). In 3 cases, the finding of the second monoclonal component was concurrent to the diagnosis of a MGUS malignant transformation to myeloma (2 cases) or Waldenström’s macroglobulinemia (1 case). Median follow-up was 23 months (12 to 252 months) for the 123 patients with BGUS. In 4 cases (3.2%) a malignant transformation was observed. Three patients developed a multiple myeloma (time to transformation was 2 years for one and 4 years for the 2 others) and one patient Waldenström’s macroglobulinemia (6 years).
Conclusions: As for monoclonal gammopathies, BGUS represent the most frequent diagnosis. However, in contrast to monoclonal gammopathies, biclonal gammopathies are more frequently associated with lymphoproliferative diseases than with multiple myeloma. The apparition of a second monoclonal component during follow up of MGUS could be associated with malignant transformation and should lead to new evaluation. The risk of transformation of BGUS seems similar to MGUS but further studies are necessary to compare the evolution of MGUS and BGUS.
Expression of Myeloma-Specific Markers in Bone Marrow Spicules Using a Novel Immunohistochemical Technique
Authors: Santra Madhumita , Shaughnessy John , Williams David R. , Barlogie Bart , Bellamy William 
Published: April 2020

Abstract
In addition to its role in diagnosis and assessing the prognosis of numerous malignancies, immunohistochemistry (IHC) is an important tool for the analysis of protein biomarker expression in tissue sections. The combination of bone marrow (BM) biopsy and IHC is frequently used to study histopathological alterations in diseases such as multiple myeloma (MM). However, due to the harsh decalcification process generally used for processing of bone marrow biopsies, protein epitopes are occasionally rendered unsuitable for IHC detection. We have developed a novel technique for processing BM spicule samples into a fibrin-clot matrix that allows for IHC detection of MM protein markers. This method does not require decalcification and results in a consistent, reliable assay. Briefly, bone marrow aspirates drawn from MM patients were subjected to a brief processing step to isolate the marrow spicules. Once isolated, the spicules were admixed with thromboplastin and pooled normal human plasma to create a clot. The clot was allowed to harden at 37°C after which it was placed into an embedding cassette for fixation and histological processing. The result was a formalin-fixed, paraffin-embedded clot suitable for IHC studies, including the construction of TMAs. Using patient paired spicule-clot and core biopsies from patients diagnosed with myeloma, we studied several antibodies including, but not limited to, kappa and lambda immunoglobulin light chains, CD138 and Cyr61, a member of the CCN family of extracellular matrix-associated signaling proteins. Results demonstrated that IHC staining for all antibodies was comparable in both the spicules and in the cores. While BM core biopsies had areas of non specific staining for several antibodies, this was not generally observed in the BM spicule samples with the exception of immunoglobulin light chains which displayed an element of non-specific staining with both samples. Moreover, we observed a consistently superior morphology in the BM spicule samples than in matched bone marrow core biopsies. In conclusion, our study demonstrates that BM spicule samples processed from MM patients using this novel fibrin-clot matrix technique were suitable for IHC and had generally lower background and non-specific staining coupled with better morphology when compared to matched core biopsies.
Effects of CK2 Inhibition on Multiple Signaling Pathways in Myeloma Cells
Authors: Piazza Francesco Alessandro , Montini Barbara , Arcidiacono Diletta , Gurrieri Carmela , Manni Sabrina , Colpo Anna , Tubi Laura Quotti , Zambello Renato , Trentin Livio , Semenzato Gianpietro 
Published: April 2020

Abstract
CK2 serine-threonine kinase plays a key role in promoting cell survival of multiple myeloma (MM) cells through modulation of extrinsic and intrinsic apoptotis and by activating pro-survival signaling such as the NF-κB and STAT3 pathways (Piazza FA et al. 2006, Blood 105:1698–1707). Here we deepened our previous results and placed the CK2 action inside critical pathways for MM cell growth, like the MAPK, STAT3 and Wnt regulated cascades.
We employed MM cell lines, primary cells from MM patients and human cervical carcinoma cell line HeLa. Inhibition of CK2 was obtained with well characterized chemical compounds (TBB, K27 and K17) or by RNA interference.
CK2 inhibition led to an increased p38 MAPK activation and to a decreased basal and growth factor stimulated-ERK1,2 phosphorylation. These changes were associated to increased activation of apoptotis and cell death. Inhibition of PP1 and PP2A phosphatases led to a decreased p38 MAPK phosphorylation upon CK2 inhibition, suggesting that CK2 may control p38 activity through the modulation of these molecules. Activation of upstream JAK tyrosine kinase upon cytokine or growth factor stimulation are a prerequisite for STAT3 activation. Following inhibition of CK2 activity, Tyk2 phosphorylation and JAK2 activation were found impaired and these effects were associated to a decreased STAT3 phosphorylation in tyr705 and ser727. In turn, activated-STAT3 nuclear migration and STAT3-dependent transcription of target genes, like the pro-survival bcl-2 family member mcl-1 and the cell-cycle promoter cyclin-D1, were significantly reduced.
Finally, Wnt activation was also affected by CK2 manipulation. Indeed, β-catenin activation and transcriptional activity were impaired by the treatment of MM cells with CK2 inhibitors; these effects were associated with reduced cell proliferation and cell death.
Altogether, these data demonstrate that CK2 is a key player placed at the cross-road of important signaling pathways for MM cell growth and reinforce the possibility that it may represent an ideal target molecule for therapeutic purposes.
Combination of DNA Methylation Inhibitor 5-Azacytidine and Arsenic Trioxide Has Synergistic Activity in Multiple Myeloma
Authors: Chen Guanghua , Lin Fengru , Depei Wu , Wang Yi , Huang Haiwen , Sun Aining , Chang Huirong , Feng Yufeng 
Published: April 2020

Abstract
This study was undertaken to study the effect of 5-azacytidine on XAF1 expression in myeloma cells and efficacy of 5-azacytidine and arsenic trioxide combination treatment in myeloma in vivo and in vitro. XAF1 expression was analyzed by semi-quantitative PCR and Western blotting. Methylation specific PCR (MSP) was used to detect methylation status of XAF1 promoter CpG islands. RPMI 8226 and XG-7 cells were treated with various concentrations of arsenic trioxide and 5-azacytidine. Expression of XAF1 mRNA variants were confirmed by gel electrophoresis and sequencing. Untreated RPMI 8226 cell expresses two variants of XAF1 mRNA. Untreated XG-7 cell has no expression of XAF1. Hypermethylation of XAF1 promoter CpG islands was detected in both cell lines. Both cell lines express full-length XAF1 mRNA transcript after treated with 2.5 μmol/L 5-azacytidine for 72 h. Our studies demonstrated that 5-azacytidine exhibits antimyeloma synergy with arsenic trioxide. 5-azacytidine treatment reversed XAF1 promoter region hypermethylation status, leading to expression of full-length XAF1 transcript. In addition, combination of 5-azacytidine and arsenic trioxide was effective in slowing myeloma growth and prolonging survival of myeloma-loaded nude mice. The findings suggested that 5-azacytidine and arsenic trioxide may be an effective combination in the therapy of myeloma patients.
Alternative Treatment for POEMS Syndrome: A Lazarus Response to Lenalidomide
Authors: Patel Tejal , Aspitia Alvaro Moreno 
Published: April 2020

Abstract
Introduction: POEMS syndrome is characterized by chronic progressive polyneuropathy (P) with constellation of other symptoms including organomegaly (O), endocrinopathy (E), serum monoclonal protein (M), and skin changes (S). Other features not included in the acronym include sclerotic bone lesions, Castleman disease, papilledema, thrombocytosis, peripheral edema, ascites, fatigue and clubbing. With limited available data, autologous peripheral blood stem cell transplant (APBSCT) is probably the mainstay of therapy in good performance status patients.
Case Description: A 37 year old male presented to the Mayo clinic with a three year history of progressive severe peripheral neuropathy leaving him wheelchair bound, as well as lymphadenopathy, hepatosplenomegaly, diabetes, proteinuria, monoclonal gammopathy and hyperpigmentation. Patient also had significant pain from diffuse sclerotic lesions, blurry vision due to papilledema and generalized edema. Based on the above findings, a diagnosis of POEMS was suspected. The diagnosis was confirmed by a biopsy of a left axillary node showing plasma cell Castleman’s disease. Patient’s bone marrow was negative for multiple myeloma and amyloidosis. Patient underwent APBSCT and had a remarkable clinical improvement that persisted over three years. Patient was able to live a normal, active life during those years.
After three years, patient developed ascites, massive splenomegaly and pancytopenia. Patient was referred again to our institution for possible splenectomy. On further evaluation, he was found to have significant portal hypertension and underwent TIPS (transjugular intrahepatic portosystemic shunt) procedure with some improvement in his symptoms. Patient remained in observation for another 18 months but returned oxygen dependent with worsening ascites, peripheral neuropathy, and encephelopathy. Further evaluation confirmed worsening POEMS. Patient’s ECOG performance status was poor therefore repeat APBSCT was not considered an option. Lenalidomide was started at 10mg/day for 21 days of a 28 day cycle with once weekly dexamethasone 40mg. The dose of lenalidomide was gradually increased to standard 25mg dose which the patient tolerated well.
By the second month patient was on therapy, his performance status improved to ECOG 1, his ascites and encephalopathy resolved, the splenomegaly improved and he no longer required oxygen. Patient has currently completed a total of 10 cycles and feels as good as 10 months post-APBSCT. The only side effect experienced by the patient is weight gain due to chronic dexamethasone therapy. Dexamethasone was weaned and discontinued after which lenalidomide continued as a single agent.
Conclusion:: Lenalidomide is a less toxic but effective alternative treatment for POEMS syndrome. The Lazarus response to the use of lenalidomide supports the principle that POEMS is a cytokine-mediated syndrome
The Incidence of Deep Vein Thrombosis Is Lower in Japanese Patients with Multiple Myeloma Treated with Thalidomide and Corticosteroids or Chemotherapy
Authors: Takei Naoko , Kami Masahiro , Tsubokura Masaharu , Miura Yuji , Issiki Takehiro , Satou Yuji , Matsue Kosei , Komatsu Tsunehiko 
Published: April 2020

Abstract
Background: Deep vein thrombosis (DVT) is an important complication of thalidomide therapy especially when it is combined with corticosteroids or chemotherapy. Japanese patients with multiple myeloma (MM) are currently prescribed thromboprophylaxis after ASCO guideline, although there are limited data for asian population. We examined the incidence of DVT in Japanese patients with MM.
Methods: We reviewed the records of 145 patients receiving thalidomide for newly diagnosed or relapsed/refractory MM at Kameda Medical Center and Tsukuba Memorial Hospital during the 10-year period 1998–2008. Patients were considered to have developed DVT if they presented with suggestive symptoms of leg swelling and had a Doppler ultra-sound study confirming acute thrombosis.
Results: 95 of the 145 patients received thalidomide in combination with a corticosteroid or other chemotherapeutic agents. 70 of 95 patients were prescribed thromboprophylaxis; of these, 53 patients received warfarin, 17 patients received aspirin during combination treatment. 38 of 70 patients were newly diagnosed. Only 1 of 70 patients (1.4%) developed DVT. None of the patients who were not received any prophylaxis developed DVT during combination treatment.
Conclusion: The incidence of deep vein thrombosis is lower in Japanese patients with MM treated with thalidomide and corticosteroids or chemotherapy than previous report from western countries.
Long Period of Complete Haematological and Molecular Remission of Chronic Eosinophilic Leukaemia after 6months Treatment with Imatinib
Authors: O’Mara Stephen K , Iland Harry 
Published: April 2020

Abstract
We report a 25 year old male patient who was diagnosed with chronic eosinophilic leukaemia based on clinical features of Skin rash, peripheral neuropathy of right hand, malaise, and hepatosplenamegaly detectable by clinical examination His blood contained 30 × 109/L Eosinophils.. A bone marrow was heavily replaced by eosinophils and eosinophil precursors. ANCA was negartive and normal Ig E levels. PCR for the PDGFR-FIP1L1 fusion gene was positive. He was initially treated with a combination of corticosteroids and alpha Interferon-2alpha and anticoagulated. At 5 months of therapy complete haematological remission had not been achieved. He was started on Imatinib mesylate 400mg per day and rapidly achieved haematological and molecular remission. Imatanib was stopped at 6 months for liver dysfunction. He was rechallenged with 100mg of imatanib after four weeks without treatment. He again developed hepatic dysfunction and imatanib was ceased completely. He remains in complete haematological and molecular remission 2 years after ceasing imatanib. He has normal performance status. This demonstrates that patients with chronic eosinophilic leukaemia who are treated with a 400mg dose of imatanib for six months may be cured or have substantial remissions. It is unclear whether the use of corticosteroids and interferon prior to the introduction of imatanib influenced the outcome. Previous published data showed that using lower doses of Imatanib does not lead to sustained remission of chronic eosinophilic leukaemia after completing drug therapy.
A Quantitive Assay and Clinical Significance for JAK2V617F Mutation in Chinese Patients with Essential Thrombocytosis
Authors: Zhang Ri , Chao Hongying , Shen Yimin , Fan Zhen , Zhang Xuhui 
Published: April 2020

Abstract
Essential thrombocytosis (ET) is a subtype of Bcr-Abl negative the chronic myeloproliferative disorders and the mutated JAK2V617F has been identified and correlated with ET. The aim of this study was to analyze the mutational status and quantitative expression of JAK2V617F mRNA transcripts in cohort of 98 Chinese patients with ET and to determine the disease features. The cohort consisted of 46 men and 52 women with median age at diagnosis was 50 years (range 17–87), granulocyte cDNA was enriched from blood samples obtained at the time of clinical assessment. A quantitative JAK2V617F measurements were made using amplification-refractory mutation sequencing polymerase chain reaction (ARMS-PCR) and capillary electrophoresis assay. Fifty-nine(60.2%) patients harboring the JAK2 V617F mutation were found, including 17 homozygous and 42 heterozygous changes. The quantitative assay by capillary electrophoresis showed that the mutated mRNA ratio was (89.9±6.7)% in the JAK2V617F homozygote and was (57.1±6.7)% in the JAK2V617F heterozygote patients, the difference was significant(P&lt;0.005). JAK2V617F-positive ET patients were older than patients without the mutation (55 versus 38 years, p&lt;0.001), and the patients whose age ≥ 60 years showed significantly higher JAK2 mutated RNA levels as compared to patients whose age ≤60 years(68.2% versus 80.2%, P =0.0056). Males and females had similar JAK2V617F prevalence (63.0% versus 57.7%) and frequency of homozygosity (17.4% versus 17.3%). As compared to heterozygous ET patients, higher leukocyte count was observed in homozygous ET patients (27.9×109/L versus 15.2×109/L, P©‚0.05), JAK2V617F-positive patients also had a higher median white cell count than wild-type patients (19.4×109/L versus 9.7×109/L, P©‚0.05), but there were no significant differences in hemoglobin concentration or platelet count. During follow-up, 30 patients suffered from documented thrombotic events, with 24 having JAK2V617F mutations. The rate of thrombotic complications in JAK2V617F-positive and homozygous patients was higher than in JAK2 wild-type and heterozygous patients, respectively. (52.2% versus 17.6%, 75% versus 32.8%, respectively ). Compared with patients without thrombotic events, there were higher levels of mutated JAK2 mRNA in patients with thrombotic events(76.9% versus 58.4%, P©‚0.05), thrombotic events were also significantly correlated with leukocytosis and older age. Our results suggest that there are different clinical features between JAK2V617F-positive and JAK2V617F-negative, homozygote and heterozygote, higher and lower levels of mutated JAK2 mRNA, and correlated with leukocytosis and thrombosis. Therefore, detection of the JAK2V617F mutational status and mutated JAK2 mRNA can help not only identify the disease state and progression, but also affect the management of ET patients.