bluuebunny/crossref_metadata_2025_split huggingface dataset:
Determination of the Maximum Tolerated Dose of Cyclophosphamide in Conjunction with High-Dose Etoposide and Carboplatin Followed by Autologous CD34+ Hematopoietic Rescue in Pediatric Patients with Recurrent/Refractory Solid Tumors.
Authors: Hudspeth Michelle P. , Duffy Barbara , Noga Stephen , Goodman Steven , Civin Curt I. , Chen Allen R. 
Published: October 2019

Abstract
Cyclophosphamide (Cy) is an alkylating agent utilized in almost all preparative regimens for hematopoietic stem cell transplantation, and in vitro sensitivity has been demonstrated with carboplatin and etoposide. Multiple previous studies have involved this combination of agents; however, they almost exclusively utilized a fixed dose of Cy and/or bolus administration. The potential fatal cardiac toxicity of Cy is likely related to high peak concentrations of active metabolites from bolus dosing. Additionally, some evidence exists that the increased metabolism of Cy with continuous infusion has a therapeutic benefit. The objective of this study was to determine the maximum tolerated dose (MTD) of continuous infusion Cy with fixed dose etoposide (2400 mg/m2) and carboplatin (2175 mg/m2) followed by autologous CD34+ hematopoietic rescue in pediatric patients with recurrent/refractory solid tumors. Twenty patients were enrolled, and fourteen patients completed therapy on study. The MTD of Cy was 60 mg/kg/24 hr by continuous 60 hr infusion on days -4 to -2 for a total dose of 150 mg/kg. Alopecia, stomatitis and nausea were the most common toxicities. The single episode of direct cardiac toxicity was reversible. Two Grade IV toxicities occurred, consisting of diarrhea in one patient and coma in another patient who recovered and remains a long-term survivor. All patients were evaluable for neutrophil engraftment, with a median time to ANC > 200/μl of 11 days (range 7–30 days). Thirteen of the fourteen patients were evaluable for platelet engraftment, with a median time to platelet count of 20,000/μl independent of transfusion of 21 days (range 11–72 days). Six out of the fourteen patients remain alive greater than five years after study completion. The MTD of Cy was lower than expected. These results suggest that autoinduction of Cy metabolism (repeated administration of Cy at 24 hour intervals leads to increased hepatic cytochrome enzymes) could be a primary factor in the determination of the MTD. Future studies should aim to further characterize the pharmacokinetics and effectiveness of continuous infusion Cy in pediatric patients.
Dose Dense Dreifach* Melphalan100 for Multiple Myeloma.
Authors: Berz David , McCormack Elise M. , Winer Eric S. , Karwan Patricia , Colvin Gerald , Rathore Ritesh , Lum Larry , Abedi Mehrdad , Elfenbein Gerald , Quesenberry Peter J. 
Published: October 2019

Abstract
Background: Tandem high dose melphalan therapy with autologous peripheral stem cell support has emerged as standard of care for patients without prohibitive comorbidities. Mucositis and gastrointestinal side effects are the most common extrahematologic side effects. Two previously published studies presented a triple transplant with a conditioning regimen melphalan 100mg/m2 (MEL100) with peripheral stem cell support every two to five months schedule. This dose reduced regimen was implemented to limit those extrahematologic dose limiting side effects in the elderly or high risk population. We are presenting a novel approach that investigates the triple melphalan100/m2 approach on a dose dense, every three weeks schedule in a patient population without significant comorbidities for conventional high dose tandem transplantation.
Patients and methods: Thirteen standard or high risk patients with stageIII multiple myeloma were prospectively treated This population contained eight patients with IgG clonality, 3 IgA, 1 nonsecretory and one light chain isotype. The induction regimens of the thirteen patients were heterogenous and included 5 VAD, 3 DCIE, 2 Thal/Dex, 2 CIE and 1 pulse decadron. Patients had stem cells harvested through peripheral blood leukopheresis, and these cells were divided into three equal sets and frozen. The patients were scheduled to receive Melphalan at 100mg/m2 on day -1, 20, and 41 and then the autologous infusions occurred at day 0, 21, and 42.
Results: All patients were able to receive all three cycles of the MEL100 regimen. Seven patients (54%) received the treatements on the every three week schedule; three treatments (23%) during the second cycle and six treatments (46%) of the third cycle had to be delayed a median of 6 and 4 days respectively. Three patients were managed completely in the outpatient setting, and the average total hospital stay for the three transplants was 18 days. Median progression free survival was 761 days (range 73 – 1571) and the overall survival of this cohort has yet to be reached. No patient had worse than grade II mucositis, and no serious adverse events were recorded.
Conclusions: Our regimen of three consecutive autologous peripheral stem cell transplants with a reduced dose of melphalan at 100mg/m2 given every three weeks is very well tolerated. The progression free survival and overall survival are similar and can be compared favorably with the standard tandem myeloma regimens. Although our data is intriguing, further studies with larger numbers need to be performed to confirm these results.
Novel Gene Delivery System of Sendai Virus Transduced Human Peripheral Monocytes.
Authors: Tanaka Tomoko , Kurita Ryo , Takasugi Kashiya , Nakamura Takafumi , Inoue Makoto , Hasegawa Mamoru , Ono Fumiko , Terao Keiji , Yoshikawa Yasuhiro , Tani Kenzaburo 
Published: October 2019

Abstract
Extensive monocytes extravasation is seen in infection sites, inflamed tissue and tumor tissues and exhibit a tissue-specific range of functions including phagocytosis, antigen presentation to T cells, and the release of a wide array of cytokines, chemokines, enzymes and nitrogen species. As monocytes have natural tropism to such tissues in each disease condition, these cells can be used as novel gene delivery system to cure the condition. On the other hand, as monocytes are quiescent cells, available gene transfer vectors are limited. Sendai virus (SeV) is a negative-strand RNA virus and has recently been used for the gene therapy vectors, SeV vectors (SeVV), for somatic gene therapy. This vector does not have potential of stable transformation of target cells because they generate no DNA intermediates and therefore are unable to integrate. Also, SeVV has been shown to ensure efficient expression of foreign genes in various types of tissues and can be used for gene transfer into monocytes. We first of all studied the gene transduction efficiency of SeVV into human peripheral blood CD 14 positive monocytes using GFP gene as a marker. About 82% of monocytes became GFP positive, whereas only 23% of lymphocytes and 1.8% of granulocytes became positive. The transduction efficiency was peaked at M.O.I.=1. The expression level was at least preserved for 24 hours in vitro. To do the in vivo preclinical experiments, we also transduced cDNA of GFP and GDNF (glial cell line-derived neurotrophic factor) into the peripheral monocytes of Macaca fascicularis monkey and high level expression of these genes were obtained. In vitro monocyte function after gene transduction are now under investigation. SeVV is considered to be promising gene transfer vector for primate monocytes.
Impact of Hydroxyurea on Thrombin Antithrombin Complex and Vaso-Occlusive Crisis in Pediatric Sickle Cell Disease.
Authors: Elalfy Mohsen Saleh , Abdelmonem Ashraf M. , Youssef Soha , Ismail Heba 
Published: October 2019

Abstract
Background: Several studies suggest that increased activity of the coagulation system may be important in the pathogenesis of vascular occlusion in sickle cell disease. Hydroxyurea (HU) has been shown to reduce the frequency of vaso-occlusive manifestations in both adults and children with sickle cell disease (SCD).
Aim: To analyze the effect of HU on Thrombin-Antithrombin (TAT) as a marker of thrombin generation and hypercoagualbility in SCD and to find out the relation between TAT level and vaso-occusive crisis.
Subjects and Method: we evaluated 37 child with sickle cell hemoglobinopathy (mean age 10.92±5.39 years) and 15 normal control children (mean age 9.75±6.34 years). Informed consent was obtained from patients and/or guardians and study approval by local IRB was obtained. Twenty-two patients (59.5%) were on HU, 15 (41.5%) patients did not receive HU, 7 (46.7%) of them were transfusion dependant. TAT assay was done in vitro using a sandwich enzyme immunoassay.
Results: Mean patients’ age at institution of HU was 8.54± 3.85 years with median treatment duration of 4.5 years. Causes for initiating HU therapy were frequent blood transfusion in 11 patients (50%), frequent pain crisis (≥ 3/year) in 9 patients (41%), severe anemia and parents refusing blood transfusion in 1 patient (4.5%) and stroke in another patient (4.5%). HU dose was 20.82±4.95 mg/kg/day. We measured TAT in all patients and compared them to healthy control. There was significant difference in TAT level in sickle cell patients (198.86±185.7) compared to healthy control 2.91±0.94, [P value < 0.0001]. When the level of TAT was compared between the HU and non-HU groups we found that patients on HU had statistically significant lower TAT level (172.36 vs.225.37) [P=0.039]. There was also a significant negative correlation between HU dose and TAT level (p=0.03). A significant positive correlation between number of vaso-occlusive crisis/year [P=0.03], frequency of pain crisis/year [P=0.04], duration of pain crisis [P=0.03] and TAT level was observed.
Conclusion: Hydroxyurea has significant inhibitory effect on thrombogenesis in sickle cell patients, which may be another mechanism for reducing vaso-occlusive crisis. Sickle cell children with higher TAT level had more frequent and severe vaso-occlusive crisis.
Impact of Clinical/Demographic Characteristics and Duration of Clinical Benefit on Average Weekly Doses of Darbepoetin Alfa and Epoetin Alfa in Oncology: Results from a Claims Analysis.
Authors: Song Xue , Long Stacey R. , Kallich Joel D. , Marder William D. 
Published: October 2019

Abstract
Background: Comparison of darbepoetin alfa (DA) and epoetin alfa (EA) has often relied on calculation of unadjusted average weekly doses (AWD). This study examined the impact of addressing differences in clinical/demographic characteristics and duration of clinical benefit (DCB) of last dose administered on AWD.
Methods: MarketScan claims data were used to build EA and DA treatment episodes among cancer patients with >= 1 EA or DA claim and continuously enrolled January–June 2005. AWDs were calculated based on 2 different DCB scenarios (DCB1 and DCB2) for the last dose administered in the episode. DCB1 assumed DA doses <=100mcg =7 days, 101–299mcg =14 days, and >=300mcg =21 days; and EA doses <35,000units =2 days and >=35,000units =7 days. DCB2 assumed DA doses <60mcg =2 days, 60–149mcg =7 days, 150–299mcg =14 days, and >=300mcg =21 days; EA doses <=15,000units =2 days, 15,001–35,000units =5 days, and >35,000units =7 days. Logistic regression examined if clinical/demographic characteristics (age, gender, cancer type, comorbidities, and metastases) influenced likelihood of DA vs EA use; ANOVA models estimated an adjusted AWD controlling for these clinical/demographic differences.
Results: 2,942 DA and 2,574 EA episodes were created. Younger patients, with more comorbidities, and advanced cancer were found to have increased likelihood of DA use (all p<0.001). The DCB analysis revealed that EA was more sensitive to DCB changes than DA. Switching from DCB1 to DCB2 increased AWD for DA from 94mcg to 105mcg, while EA AWD decreased from 44,044units to 35,496units. With ANOVA adjustment to equalize the populations, estimated AWD for DCB1 and DCB2, respectively, were 97mcg and 104mcg for DA and 41,902units and 34,973units for EA.
Conclusions: Equalizing clinical/demographic characteristics and addressing possible DCB differences for DA and EA can substantially change the average weekly doses calculated from claims data; unadjusted AWDs may be misleading. In addition, comparisons of dose using claims data must be interpreted with caution because such data lack information on patient clinical outcomes of erythropoiesis stimulating agents and are subject to bias including data recording errors and confounding by indication.
Efficacy, Safety and Tolerability of a Novel High-Purity, Double Virus-Inactivated VWF/FVIII-Concentrate in the Treatment of Von Willebrand Disease (VWD) - Clinical Experience in the Treatment of Bleeding and Surgery.
Authors: Schwartz B.A. , Windyga J. , von Depka M. , Walter O. , Jansen M. , Auerswald G. 
Published: October 2019

Abstract
Introduction: Wilate is a highly purified, double viral inactivated, VWF/FVIII-concentrate developed for the treatment of VWD. A program of prospective studies was conducted in VWD patients to investigate the clinical efficacy and safety of Wilate® in acute bleeding episodes, prophylaxis and surgical procedures.
Methods: In these prospective open label studies 70 VWD patients (37 VWD of type 3) were treated with the VWF/FVIII concentrate Wilate and followed to assess clinical efficacy and tolerability. Dosing and monitoring were performed using FVIII assays. This is standard practice in most of the study sites and was feasible because of the product’s physiological 1:1 ratio and parallel PK profiles of FVIII and VWF. Dosing decisions were based on established guidelines and the investigators’ clinical judgment. The efficacy of the product was rated on a four-point scale by the investigator.
Results: Bleeding- 43 patients (25 of type 3) were treated for a total number of 1088 bleeding episodes (BE). The median dose per exposure day (ED) was calculated to be 31 IU VWF/FVIII:C/kg and were treated for a median of 1.2 days, with excellent or good efficacy achieved in 96% of treatments. Surgery- Efficacy and safety was also studied in 31 VWD patients who underwent 54 elective or urgent surgical procedures of which 27 were classified as major; the overall efficacy was rated as excellent or good in most cases (94%), with a mean dose per infusion being 34 IU VWF/FVIII:C/kg. Pediatric Use- A total of 8 children below 12 years of age with 310 BE s were treated, with an excellent/good efficacy in 98% of bleedings. Prophylaxis- 19 patients were on a prophylactic regimen for more than 10 consecutive weeks (total of 2,338 ED), with an overall reduction of bleeding frequency. Tolerability - Out of 5,662 rated infusions in all studies, the tolerability was assessed as “very good” or “good” in 99% of the cases.
Conclusions: The results of the prospective clinical trial program demonstrate the safety, efficacy and tolerability of this VWF/FVIII concentrate for the treatment of acute bleeding episodes, prophylaxis and surgical procedures in patients with VWD.
Tonsillar CD38+ Activated B Cell Fraction Harbours the Highest EBV Load in Patients with Infectious Mononucleosis.
Authors: Chaganti Sridhar , Bell Andrew , Heath Emily , Bergler Wolfgang , Kuo Michael , Starczynski Jane , Rudzki Zbigniew , Rickinson Alan 
Published: October 2019

Abstract
Epstein-Barr virus (EBV) persists within the B cell system and, in the blood of healthy virus carriers, is sequestered in the immunoglobulin (Ig)Dneg, CD27+ (“class-switched”) memory B cell compartment. This selective colonisation is apparent even in the blood of infectious mononucleosis (IM) patients undergoing primary EBV infection. Exactly how this is achieved is not understood, one theory being that the virus infects naïve B cells preferentially in vivo and drives them into memory using the physiologic process of germinal centre (GC) transit within lymphoid tissues. Here we have studied the distribution of EBV-infected B cells in tonsil cell preparations from acute IM patients and from chronic carriers. The following subsets of tonsillar B cells were isolated to high purity by FACS sorting and their EBV DNA loads were then determined by quantitative PCR: Naïve (CD38neg, IgD+, CD27neg); Class-switched memory (CD38neg, IgDneg, CD27+); Non-switched memory (CD38neg, IgD+, CD27+); Germinal centre (CD38+); and Plasma cell (CD38hi). In 15 carrier tonsils, the highest EBV load was consistently detected in class-switched memory, followed by significant amounts in non-switched memory. However, viral load in the CD38+ GC subset was usually very low, comparable to that in the naïve B cell fraction. This contrasted with the situation in 7 IM tonsils where overall viral loads were on average 100-fold higher than above. Here most viral DNA was consistently found in the CD38+ fraction, with lower loads in the class-switched memory and the smaller non-class switched memory subsets. We then investigated further the phenotype of the CD38+ population and found that, in IM tonsils, this not only included cells with the CD10 and CD77 markers of germinal centre origin but also other cells lacking these markers. Adopting a different FACS sorting strategy based on CD10, CD77 and IgD staining, we found that the virus was not present at significant levels in either the centrocyte (CD10+ CD77neg) or centroblast (CD77+) fractions. Thus EBV appeared to be concentrated in B cells expressing CD38 as a marker of activation rather than of germinal centre origin; this concurred with the results of immunohistochemical staining for the EBER RNAs showing an extrafollicular distribution of virus-infected cells in these IM tonsils. Our findings suggest that CD38 expression is a useful marker for B cells recently activated by EBV infection in vivo and show that, during primary EBV infection, the tonsillar CD38+ B cells lacking germinal centre markers harbour the highest viral load. If EBV does exploit the germinal centre pathway as an entry route into B cell memory, where primary infection is manifest as IM this pathway must be taken by a very small fraction of infected B cells and/or must have been completed before disease symptoms appear.
Complementary JAK/STAT Signalling Is Required for the Pro-Inflammatory Effects of CD40 Ligation: Differential Effects in Human Myeloid and B Cells.
Authors: Conzelmann Michael , Rodionova Elena , Hess Michael , Giese Thomas , Ho Anthony D. , Dreger Peter , Luft Thomas 
Published: October 2019

Abstract
CD40L represents a strong endogenous danger signal that induces pro-inflammatory activation of CD40-expressing cells such as dendritic cells (DC), monocytes, and B cells. However, since CD40 activation alone is insufficient to induce pro-inflammatory cytokines such as IL-12p70, we studied whether CD40-mediated pro-inflammatory activity might be dependent on co-signalling pathways involving JAK/STAT. Using quantitative Western blotting, we demonstrate that JAK/STAT signalling is induced by cytokines such as IL-4, GM-CSF and IFNg, whereas CD40 activation mediates NFkB signalling. CD40L-induced IL-12p70 and IL-10 secretion in human DC, monocytes, B cells, and chronic lymphocytic leukemia (CLL) cells was measured upon complementary JAK/STAT activation by IL-4, GM-CSF and IFNg in the presence and absence of specific inhibitors of JAK2, JAK3, and pan-JAK. Whereas IL-12p70 could not be induced by CD40 ligation or by cytokines alone, IL-12p70 secretion and suppression of IL-10 was reproducibly observed after co-stimulation of CD40L with IL-4, GM-CSF, or IFNg. This effect could be completely reversed by pan-JAK inhibition. Persistence of IL-4/GM-CSF/IFNg-mediated JAK/STAT signalling as late as 12 hours following cellular activation via CD40 was required for IL-12p70 secretion as shown by the effects of delayed JAK inhibition. Similarly, persistence between 12 and 24 hours of IL-12p35 and p40 mRNA expression correlated best with the level of IL-12p70 secretion. Specific inhibition of JAK2 and JAK3 further revealed a context-dependent action of the distinct JAK family members: JAK2 showed a strong co-dominant effect in the setting of IL-4-induced JAK/STAT activity. Both, JAK2 and JAK3 were required for IL-12p70 secretion, whereas JAK2 alone was sufficient to modulate IL-10 secretion. However, in the context of IFNg-induced JAK/STAT signalling in DC, neither JAK2 nor JAK3 inhibition had effects on IL-12p70. Here, only inhibition by the pan-JAK inhibitor involving JAK1 abrogated IL-12p70 secretion, indicating that in IFNg-dependent signalling, JAK2 is apparently sub-dominant to JAK1 and had only a small enhancing effect on IL-10. This context dependence markedly differed in myeloid cells and B cells, as normal and malignant (CLL) B cells maintain a co-dominant JAK2 activity in the context of IFNg-induced JAK/STAT-signalling. In conclusion, complementary JAK/STAT signalling is required for the pro-inflammatory effects of CD40 ligation in humans, with different JAK subset predominance in myeloid and B cells. These results may open new ways of lineage-specific interfering with CD40 signals by modulating JAK/STAT activity using tyrosine kinase inhibitors.
Acquisition of New In Vivo Genomic Change over Time in Multiple Myeloma.
Authors: Tai Yu-Tzu , Buon Leutz , King Benjamin , Li Cheng Li Cheng , Burger Peter Burger Peter , Li Xian-Feng , Song Weihua , Schlossman Robert , Richardson Paul , Anderson Kenneth C. , Munshi Nikhil 
Published: October 2019

Abstract
Multiple myeloma (MM) cells present with significant genetic abnormality. This genetic instability is considered responsible for not only development of malignant phenotype but also progression of myeloma as well as development of drug resistance. We have previously demonstrated that MM cell lines have elevated homologous recombination activity that leads to acquisition of new genomic changes over time and is associated with development of drug resistance (Blood2004; 104: 3409). However, such genomic evolution in patient samples has not been documented. Here, we have performed a genome wide loss of heterozygosity (LOH) assay, using high-density oligonucleotide arrays capable of measuring up to 500K single nucleotide polymorphisms (SNP) loci, and identify areas of amplification and losses to determine ongoing development of new changes that may reflect instability. We have evaluated 17 MM patients with purified myeloma cells obtained at two time points, at least six months apart. CD138–expressing myeloma cells from these patients were purified and their peripheral blood mononuclear cells were also obtained. Genomic DNA from these cells was digested with StyI, PCR amplified and hybridized to 250K SNP array. Results from CD138+ myeloma cells from two time points or more were compared with each other using the dChip software for LOH and copy number analysis. During the LOH analysis, we have adapted a tumor vs. tumor comparison to track all the new SNPs affected. We have observed that myeloma cells acquired new LOH loci in the subsequent samples. These new areas of genetic changes were recurrent; for example on chromosome 13, an average area of recurrent LOH of 87 Mb was found to be present among three patients. We were able to observe recurrence of significant copy changes on several chromosomes with/without LOH. The reproducible area of new acquisition of LOH and/or significant copy changes interestingly involved areas on chromosomes 1p, 1q, 8p, 9q, 13q, 15, 16q, 20p, 21q, and X that may have significant role in the pathogenesis and progression of the disease. We are currently analyzing the gene expression profile from these time points to identify expression changes correlating with the observed genomic changes on follow up samples. In summary we demonstrate continued acquisition of new genomic changes in vivo in MM over time and provide a molecular basis for evolution of more aggressive disease able to escape therapeutic interventions. The mechanism governing such evolution is an important target to not only inhibit progression of the disease but also development of drug resistance.