bluuebunny/crossref_metadata_2025_split huggingface dataset:
Construction of Wif1-IgG1/Fc Recombinant Vector and Characterization of Its Expression in CHO Cells.
Authors: Cai Zhen , Hu Jie 
Published: October 2019

Abstract
Wnt signalling dysregulation has been implicated in cancers, including lymphoma. The Initiation of Wnt signalling modulated by soluble Wnt ant agonists, including soluble frizzled related proteins, dickkopf proteins, and Wnt inhibitory factor-1 (Wif1). These natural inhibitors can be used directly as theraputic reagents for cancers. But the recombinant antagonist only give very low blood levels, are cleared rapidly from the body (only several hours) and must be administered daily via subcutaneous injection. Therefore, development of new forms of Wnts inhibitors with prolonged circulating half-life is of significant interest. Human IgG immunoglobulins have longer circulating half-life (21 days in humans). When fused with a Wnt inhibitor, its dimeric structure further increases the effective size and circulating half-life. Thus administration of vector plasmid may be every 2~3 week. Moreover, IgG Fc can interact with FcR of CTL, NK, MΦ, and complements and the target cells can be killed through ADCC and CDCC. We constructed a Wif1-IgG1/Fc expressing vector and expressed it in Chinese hamster ovum cells in order to seek effective methods to inhibit lymphoma growth. Wif1 and IgG/Fc cDNAs were amplified by RT-PCR from human lymphocyte and cloned into the eukaryotic expression vector pcDNA3.0 by direct cloning and the resultant recombinant plasmid pcDNA Wif1-IgG1/Fc was transfected into 293 cells with lipofectmin. The constitutively expressing cells were obtained by G418 screening. Expression of the fusion protein was confirmed by Western blot, the fusion protein was purified with affinity chromatography, and its’ biological activity was examined by flow cytometry, MTT colorimetry and ELISA. Our results show that the fusion protein significantly upregulated Wif1 density and promoted molt3 cell apoptosis, which was time dependent. The molt3 cell apoptosis induced by the fusion protein increased in the presence of NK cells or complements. Thus, we have successfully constructed and functionally expressed Wif1-IgG1/Fc fusion protein and shown that the purified fusion protein could kill molt3 cells through apoptosis and by ADCC and CDCC.
Stem Cell Transplantation in Children with Infantile Osteopetrosis Is Associated with a High Incidence of VOD, Which Could Be Prevented with Defibrotide.
Authors: Corbacioglu Selim , Manfred Hoenig , Lahr Georgia , Stoehr Susanne , Berry Gaurav , Friedrich Wilhelm , Schulz Ansgar S. 
Published: October 2019

Abstract
Background: Malignant infantile osteopetrosis (MIOP) is a rare hereditary disorder of osteoclast function, which can be reversed by hematopoietic stem cell transplantation (SCT). In recent studies defibrotide (DF) demonstrated superior efficacy in the treatment of hepatic veno-occlusive disease (VOD).
Methods: Twenty children with MIOP were consecutively transplanted in our centre between 1996 and 2005. Eleven of these patients were transplanted between 1996 and 2001 without any VOD prophylaxis. Thereafter nine patients were transplanted between 2001 and 2005 with prophylactic DF from the first day of conditioning until 30 days after SCT because we observed a high incidence of VOD in transplanted patients and explored the prevention of this complication by using DF as a prophylaxis.
Results: Patients without DF experienced an overall incidence of VOD of 63.6% (7/11). VOD was severe in three patients and one patient succumbed to VOD related multiorgan failure (MOF). In nine patients consecutively transplanted between 2001 and 2005 only one patient (11.1%) was diagnosed with moderate VOD.
Conclusion: SCT in patients with MIOP is associated with a very high risk to develop VOD. Prophylactic DF was implemented in our current transplant protocol and reduced the VOD rate dramatically in this high risk population.
Alemtuzumab in the Treatment of Steroid Refractory Acute Graft-Versus-Host Disease.
Authors: Gomez-Almaguer David , Ruiz-Arguelles Guillermo , Gonzalez-Llano Oscar , Cantu Olga G. , Gutierrez Cesar H. , Jaime-Perez Jose Carlos , Tarin-Arzaga Luz , Giralt Sergio 
Published: October 2019

Abstract
Corticosteroids therapy is the mainstay of treatment for GVHD, however, it heavily impacts on post transplant morbidity and new modalities are continually needed. Alemtuzumab a humanized monoclonal antibody to CD52 has been used mainly as GVHD prophylaxis. Only a few patients have been treated with this antibody. From December 2004 to May 2006, we recruited 13 steroid refractory acute GvHD patients in a prospective trial evaluating the efficacy of alemtuzumab (Campath 1H) after exclusion of other severe HST-related complications. Primary endpoints were response to treatment after 14 and 28 days. Secondary endpoints were side effects and incidence of infectious complications. Treatment consisted of Campath 1H 10mg given s.c. on days 1–5. Median age was 33 years old (range:1–59) years, a fludarabine-based reduce intensive conditioning (RIC) regimen was used and the hematopoietic cells were obtained from HLA-identical siblings in 12 cases and one patient received stem cell from umbilical cord blood. All but one received CSA and MTX for GvHD prophylaxis. GvHD affected gut in 6 cases, skin in 3 liver in 4, and combination of gut and skin in 6 patients. In 6 of the 13 patients the clinical manifestations of GVHD were noticed after the first 100 days of HSCT. Complete resolution of GvHD, partial response and no response were seen respectively in 23%, 62% and 15%. Six over the 13 patients were able to decrease steroid use. Five patients developed CMV (pp65) reactivation and 3 of them were successfully treated with valganciclovir. All patients maintained complete chimerism during and after alemtuzumab therapy, and after a median follow-up of 4 months (range, 1– 17months), 8 remain alive, 3 without evidence of GVHD. Five patients died, 3 due to GvHD and the others due to infectious complications. This preliminary study suggests that alemtuzumab is a well-tolerated agent and has a beneficial effect in the treatment of refractory GvHD. It is only a pilot study and more studies are needed, but we suggest that this modality could be used early in the management of these patients in order to improve quality of life and reduce the long-term side effects of corticosteroids.
Hepatic Injury Following Reduced-Intensity Unrelated Cord Blood Transplantation for Adult Patients with Hematological Diseases.
Authors: Kusumi Eiji , Kami Masahiro , Kanda Yoshinobu , Seki Kunihiko , Fujiwara Masayo , Koyama Rikako , Komatsu Tsunehiko , Kodama Yuko , Matsumura Tomoko , Yuji Koichiro , Miyakoshi Shigesaburo , Taniguchi Shuichi 
Published: October 2019

Abstract
Liver injury is a common complication in allogeneic hematopoietic stem cell transplantation. Its major causes comprise graft-versus-host disease (GVHD), infection, and toxicities of preparative regimens and immunosuppressants; however, we have little information on liver injuries following reduced-intensity cord blood transplantation (RICBT). We reviewed medical records of 104 recipients who underwent RICBT between March 2002 and May 2004 at Toranomon Hospital. Preparative regimen and GVHD prophylaxis comprised fludarabine/melphalan/total body irradiation, and cyclosporine or tacrolimus. We assessed the etiology of liver injuries based on the clinical presentation, laboratory results, comorbid events, imaging studies in 85 patients who achieved primary engraftment. The severity of liver dysfunction was assessed according to the National Cancer Institute Common Toxicity Criteria version 2.0. Hyperbilirubinemia was graded according to the report by Hogan et al. (Blood.2004;103:78–84). Moderate to very severe liver injuries were observed in 36 patients. Their causes included cholestatic liver disease related to GVHD or sepsis (CLD) (n=15), GVHD (n=7), cholangitis lenta (n=5) and others (n=9). Median onsets of CLD, GVHD and cholangitis lenta were day 37, 40 and 22, respectively. Frequencies of grade 3–4 ALT elevation were comparable among the three types of hepatic injuries. Serum GTP was not elevated in any patients with cholangitis lenta, while 27% and 40% of patients with CLD and GVHD, respectively, developed grade 3–4 GTP elevation. Multivariate analysis identified two risk factors for hyperbilirubinemia; grade II–IV acute GVHD (relative risk 2.23, 95% confidential interval (95% CI) 1.11–4.47, p=0.024) and blood stream infection (relative risk 3.77, 95% CI 1.91–7.44, p=0.00013). In conclusion, the present study has demonstrated that the hepatic injuries are significant problems following RICBT, and that GVHD and blood stream infection contribute to their pathogenesis.
The Relationship between Anemia and Chronic Kidney Disease (CKD) in Older Nursing Facility (NF) Residents.
Authors: Artz Andrew , Robinson Bruce , Culleton Bruce , Critchlow Cathy , Sciarra Angela , Audhya Paul 
Published: October 2019

Abstract
Anemia is a common condition among older NF residents that often has unidentified causes. CKD is common in older adults and frequently contributes to anemia. However, the association between declining kidney function and the prevalence and severity of anemia in NF residents has not been well characterized. We retrospectively analyzed the independent association of CKD and anemia among older residents in 372 NFs across the US. Any resident who was admitted to a NF between 01 Jan 2002 and 31 Dec 2003, was still active as of 31 Jan 2004, was ≥ 65 years of age, and had more than one serum creatinine (SCr) measurement and more than one hemoglobin (Hb) documented during the admittance period, was considered. Residents who had end-stage disease, were comatose, or were receiving dialysis were excluded. CKD was conservatively defined as an estimated glomerular filtration rate (eGFR) < 60 mL/min/1.73 m2 (MDRD equation). Anemia was defined using the WHO criteria for anemia (< 13 g/dL for men and < 12 g/dL for women). Regression analysis was used to determine the association between kidney function and Hb levels after adjusting for age, race, and gender. Of the 10315 residents identified, 6200 were eligible for this study: 85% (5291/6200) were white, 70% (4354/6200) were female, and the mean ± SD age was 83.1 ± 7.9 years. Anemia was found in 59.6% (3697/6200) residents as defined by the WHO criteria and CKD was present in 43.1% (2671/6200) residents. The odds of having anemia was 1.47 times greater (95% CI: 1.33, 1.63) for residents with CKD than those without CKD. The severity of anemia also increased with the presence and severity of CKD. The odds ratio increased to 1.65 (95% CI: 1.49, 1.84) after adjusting for age, race, and gender. Of those with CKD, 36.4% (972/2671) had a Hb < 11 g/dL, however, only 5.1% (50/972) and 25.4% (247/972) received erythropoiesis-stimulating protein (ESP) and iron therapies, respectively. Our data suggest CKD contributes to the high prevalence of anemia in older NF residents. These results reinforce the importance of evaluating kidney function and Hb level in older NF residents. Future studies evaluating the impact of anemia and benefits of anemia therapy are warranted in this setting.
Timing of Second Autologous Transplantations in Multiple Myeloma: Results of a Multicenter Sequential Randomized Clinical Trial.
Authors: Abdelkefi Abderrahman , Ladeb Saloua , Ben Othman Tarek , Torjman Lamia , Lakhal Amel , Ben Romdhane Neila , El Omri Halima , Elloumi Moez , Bellaj Hatem , Jeddi Ramzi , Aissaoui Lamia , Msadek Fahmi , Saad Ali , Hsairi Mohamed , Boukef Kamel , Amouri Ahlem , Louzir Hechmi , Dellagi Koussay , Ben Abdeladhim Abdeladhim 
Published: October 2019

Abstract
Background: Autologous stem cell transplantation (ASCT) is now considered standard therapy in young patients (<65 years) with multiple myeloma (MM). The Intergroupe Francophone du Myelome conducted a randomized trial of the treatment of MM with high-dose chemotherapy followed by either one or two successive ASCTs. The probabilities of event-free-survival and overall survival were doubled with a double transplant. However, no randomized trial has compared tandem transplant up-front with a strategy including planned second ASCT at relapse or progression. Therefore, we performed a multicenter, sequential, randomized trial designed to assess the optimal timing of a second ASCT.
Methods: From May 2003 to April 2006, 140 patients with symptomatic MM (de novo) and less than 60 years of age, were randomly assigned to receive either tandem transplantation up-front (within 6 months of the first transplantation) [Arm A, n=69] or one ASCT followed by a consolidation therapy with thalidomide (day +90, 100 mg/per day during 5 months) [Arm B, n=71]. Patients included in the arm B received a second transplant in case of disease progression on consolidation therapy, or in case of relapse in responders. Clinical characteristics of each group were similar. In both arms of the study, ASCT was preceded by first-line therapy with thalidomide-dexamethasone and subsequent collection of peripheral blood stem cells with high-dose cyclophosphamide (4 g/m2) and G-CSF. Data were analyzed on an intent-to-treat basis.
Results: With a median follow-up of 23 months (range: 6–34), the 2-year overall survival was 55% in the arm A and 75% in the arm B. Survival curves were not different (P=0.28, log-rank test). The 2-year event-free survival was 41% in the arm A and 60% in the arm B (P=0.4, log-rank-test). In the arm B, relapse-free survival of ≥ 16 months following the first transplantation was an important predictor of overall survival (p< 0.001).
Conclusion: Data from the present study suggest that up-front single ASCT followed by a consolidation therapy with thalidomide and a second ASCT after relapse or progression is a safe and effective global strategy to treat MM patients. Longer follow-up is needed before definite conclusions can be given concerning the optimal timing of second autologous transplantations in patients with MM.
A Humanized Anti-IGF-1R Monoclonal Antibody (AVE1642) Synergizes with Bortezomib To Induce Apoptosis in CD45neg Myeloma Cells.
Authors: Descamps Geraldine , Wuilleme-Toumi Soraya , Gomez-Bougie Patricia , Venot Corinne , Bladt Friedhelm Bladt , Bataille Regis , Amiot Martine 
Published: October 2019

Abstract
IGF-1 is a well-known growth factor for human myeloma cells. Thus, therapeutic strategies targeting IGF-1R have been proposed for the treatment of multiple myeloma. In this study, we investigated the effect of a humanized antagonistic monoclonal antibody to the IGF-1R (AVE1642). We showed that AVE1642 selectively inhibits IGF-1R but not insulin signaling in human myeloma cell lines (HMCL). Since we had previously shown the functional relevance of CD45 expression in the growth of myeloma cells, and the association of CD45 negative (CD45neg) status with a less favorable clinical outcome, both CD45 positive (CD45pos) and CD45neg HMCL were studied. We found that AVE1642 strongly inhibited the growth of CD45neg HMCL, leading to a G1 growth arrest, whereas it had almost no effect on the growth of CD45pos HMCL. Indeed, by cell counting over four days in standard culture conditions, AVE1642 induced a significant growth inhibition of the CD45neg LP-1 HMCL of 90% whereas it had no effect on the growth of the CD45pos XG-1 HMCL. Furthermore, AVE1642 binding induced a significant reduction of IGF-1R expression. In western blot, a decrease of 60% the IGF-1R expression of the LP-1 CD45neg HMCL was observed. This decrease was also confirmed by flow cytometry analysis. Finally, a stable CD45 silencing XG-1 cell line was generated and became sensitive to AVE1642. By cell counting over four days in standard culture conditions, AVE1642 inhibited the growth of the shCD45neg XG-1 HMCL of 56% whereas it had almost no effect on the growth of the shLuci-XG-1 HMCL. Thus, for the first time, we provided direct evidence that the expression of CD45 renders myeloma cells resistant to IGF1R inhibition. Furthermore, AVE1642 in combination with Bortezomib strongly increased the apoptosis induced by Bortezomib alone on CD45negHMCL. The CD45neg LP-1 HMCL was considered as a modestly sensitive cell line since 10nM of Bortezomib, which is in the range of therapeutic concentrations, induced 40±16% of apoptosis. Bortezomib in combination with AVE1642 was found to induce 80±1% of apoptosis in the CD45neg LP-1 HMCL. Apoptosis induction by AVE1642/Bortezomib combination was associated with an important induction of Noxa, a BH3-only pro-apoptotic protein that was increased by the addition of AVE1642. We also found a strong increase of caspase-3, -8 activation under this combination compared to Bortezomib alone. Taken together, these results support that therapy directed against IGF-1R associated with Bortezomib, could be beneficial in treating CD45neg patients, a population known for poor clinical outcome.
Transcriptional Changes Induced by Imatinib and Nilotinib in the Chronic Myelogenous Leukemia (CML) Cell Line K562.
Authors: Neumann Frank , Bruennert Daniela C. , Koch Anne-Marie , Bruns Ingmar , Gattermann Norbert , Kronenwett Ralf , Haas Rainer 
Published: October 2019

Abstract
Introduction: Nilotinib is a selective bcr-abl tyrosine kinase inhibitor that is 30-fold more potent than Imatinib in vitro. To examine the molecular and functional effects of Nilotinib and Imatinib we performed gene expression and functional analyses in K562 cells following in vitro treatment with the two tyrosine kinase inhibitors. Particular emphasis was put on 1539 genes which we found to be differentially expressed in primary CD34+ cells from patients with CML in first chronic phase in comparison to CD34+ cells from normal bone marrow (Diaz-Blanco et al., Leukemia 2006).
Methods: Affymetrix U133A 2.0 microarrays covering 21.722 probe sets were used to analyse the gene expression profile of 5x107 K562 cells after 24h in vitro treatment with Imatinib (0.5 μM) or Nilotinib (0.05 μM) (half maximal inhibitory concentration, IC 50). FISH analysis confirmed the K562 cell line to be BCR-ABL positive. Gene expression data of the treated cells were compared with the data of untreated cells. In addition, proliferation (Cell Titer 96 AQueous One Solution Cell Proliferation Assay, Promega), apoptosis (Cell Death Detection ELISAPLUS, Roche) and cell cycle (FITC BrdU Flow Kit, BD Pharmingen) assays were performed. A colony assay was performed to see differences in cell growth.
Results: Looking at those 1539 differentially expressed genes in K562 cells which distinguish patients with CML from healthy donors, we found that Imatinib led to a significant downregulation of 187 and upregulation of 45 genes. In general, Nilotinib had a more pronounced effect than Imatinib regarding the number of genes affected and the degree of suppression. It caused downregulation of 418 and upregulation of 41 genes. Of note, genes affected by Nilotinib included all genes altered by Imatinib such as those related to bcr-abl signalling (Lyn, BCL2, Myc, PIK3CB, G3BP2). Downregulation of genes involved in cell cycle (CDK2, ORC5L, MCM3, POLE2, CCNG1) was only observed following Nilotinib exposure. The stronger effect of Nilotinib is in line with the results of cell cycle experiments showing that Nilotinib exposed cells had the lowest proportion of actively cycling cells. The proportion of apoptotic K562 cells was 5.5 fold greater following treatment with Nilotinib in comparison to Imatinib after 24 hours. Treatment with either Imatinib or Nilotinib produced a similar apoptotic rate and similar decrease in cell numbers after 96 hours. In the colony forming assay, the controls (K562 cells incubated with DMSO only) displayed strong leukemic growth which was inhibited by both Nilotinib and Imatinib, allowing only small clusters to appear.
Conclusion: Nilotinib is apparently more potent than Imatinib with regard to the number of genes downregulated and the degree of their suppression. Many of the suppressed genes are associated with bcr-abl signalling and cell cycle.
High-Resolution Assessment of Gains and Losses of Chromosomes in Patients with Multiple Myeloma Treated with Bortezomib.
Authors: Danaee Hadi , Bryant Barb , Roels Steve , Lichter Dave , Koening Eric , Trepicchio William L. , Boral Anthony , Richardson Paul , Meng Yan , Bergsagel Peter Leif , Sonneveld Pieter , Broyl Annemiek , Mitsiades Constantine , Chng Wee-Joo , Anderson Kenneth , Mulligan George 
Published: October 2019

Abstract
Multiple myeloma is a disease of the antibody producing plasma cells in the bone marrow. It has been characterized by frequent genomic alterations including gains and losses of chromosomes. Some of these alterations result in poor clinical prognosis. Bortezomib, a first in class proteasome inhibitor, is approved for the treatment of relapsed multiple myeloma. To better understand how the genome wide changes observed in myeloma relate to prognosis and treatment response, we have used single nucleotide polymorphism (SNP) array technology to assess genotypes and DNA copy number at up to 100,000 genomic loci. DNA from 147 bone marrow tumor biopsies collected in multi-center phase II and III clinical trials of relapsed multiple myeloma patients prior to treatment with Bortezomib were hybridized on SNP arrays to assess genomic aberrations. Commonly seen genomic alterations in myeloma were observed, including deletions of chromosome 13 (37%), 1p (25%), 6q (23%), and amplifications on 1q (35%). We found that 35% of this relapsed myeloma population exhibited a hyperdiploid genome characterized by gains of chromosomes 3, 5, 7, 9, 11, 15, and 19. Other notable genomic deletions included 17p (22%), 8p (27%) and 10p (16%). A correlation of response to bortezomib therapy in patients with differential gains and losses at the chromosomal level is ongoing. Consistent with previous reports, response to bortezomib was observed in patients with poor prognostic chr 13 deletions. A more in-depth analysis of specific loci associated with clinical outcome will be discussed. In summary, this data will help further define genomic aberrations in multiple myeloma and their association with treatment outcome.