bluuebunny/crossref_metadata_2025_split huggingface dataset:
Predicitve Value of Pretransplant Testing.
Authors: Copelan. Edward A. 
Published: April 2020

Abstract
Routine evaluation prior to stem cell transplantation generally includes assessment of left ventricular ejection fraction (EF), pulmonary function, renal function, liver function, and functional status. Patients in whom compromised organ function is detected, are often deemed ineligible for myeloablative transplantation. However, the predictive value of specific pretransplant tests is not well documented, and has not been critically analyzed in patients receiving busulfan-based preparative regimens. We analyzed the prognostic value of the pretransplant evaluation performed in 1035 consecutive patients who underwent autologous (n=444) or allogeneic (n=591) transplantation with busulfan-based regimens between 2/84 and 12/03 for acute (250) or chronic leukemia (198), myelodysplasia (50), multiple myeloma (88), or Hodgkin’s (139) or non-Hodgkin’s lymphoma (303). Ages ranged from 4–75 (median 41). BuCy was given to 477 and BuCyVP16 to 558 individuals.
Ninety three patients had EF &lt; 50%, 61 had DLCO &lt; 65%, 103 had elevated creatine, 130 had elevated tranaminases or bilirubin, and 116 had Karnofsky scores &lt; 80%. For the entire group, multivariable analysis demonstrated only Karnofsky score &lt; 80% (P &lt; .001) and elevated LFTs (P=.04) to have significant adverse prognostic influence on survival. Karnofsky score was significant in patients undergoing autologous (P &lt;.001) or allogenic (P &lt;.001) transplantation. Neither EF nor DLCO were predictive of poor outcome for the overall group (P &gt;.2), nor for the autologous or allogeneic transplant groups. However in patients undergoing transplants from unrelated donors (n=119) DLCO &lt; 65% was a significant adverse prognostic factor (P=.004). There was no association of compromised cardiac or pulmonary function with cardiac or pulmonary failure as a primary cause of death. This study suggests that moderately compromised organ function, particularly cardiopulmonary limitations which might not be clinically apparent, are of limited value in prediction of transplant outcome. Direct clinical assessment, including functional status are of much greater value in prediction of outcome. It is hoped that this investigation will stimulate larger studies of pretransplant predictors of outcome with attention to their relevance to specific preparative regimens, sources of stem cells, and other treatment variables.
Endothelial Cell Injury of Carmustine (BCNU) in Hepatic Sinusoids: Contribution of Oxidation and Immune Reaction.
Authors: Ozdemir Oner , Ravindranath Yaddanapudi , Savasan Sureyya 
Published: April 2020

Abstract
Background: Carmustine (BCNU) is used for the treatment of brain tumors and is included in conditioning regimens of stem cell transplantation. It has been long believed that BCNU causes hepatotoxicity by inducing pericholangitis and intrahepatic cholestasis leading to biliary cirrhosis. Either hepatocytes or biliary ductal cells have been thought to be the target in BCNU-induced liver toxicity. However, the mechanisms operational at cell level have not been well elucidated. Possible explanations include glutathione depletion in cells, DNA damage accompanied by cellular oxidative stress and/or acceleration of apoptosis. It has been shown that BCNU treatment inhibits hepatic glutathione reductase activity in rats. Recently, a two-step process involving free radical formation and immune reaction has been proposed as a mechanism of BCNU-induced alveolar epithelial cell damage by our group (Savasan S, Ozdemir O, Buck S, et al., 95th annual meeting of the AACR, Orlando, March 27–31, 2004. Abstract#: 1133, 2004). In this study, we have investigated BCNU effect on human liver sinusoid endothelial cells (HLS-EC) in vitro.
Material and Methods: Cytotoxicity in HLS-EC was measured by flow cytometric detection of annexin V/PI-positive cells, DiOC6(3) /PI staining, further quantification of fluorosphere-adjusted events and trypan blue exclusion test in repeated experiments. The generation of reactive oxygen species (ROS) was measured by flow cytometric DCF staining. Lymphokine activated killer (LAK) cell-mediated cytotoxicity against BCNU-treated and untreated HLS-EC was measured by flow cytometric cell mediated cytotoxicity assay. The influence ROS-scavenging compound butylated hydroxyanisole (BHA) and anti-inflammatory dexamethasone (DXM) on BCNU treatment was also studied.
Results: Severe HLS-EC damage occurred following BCNU treatment shown by four different cytotoxicity methods. This effect was associated with oxidative stress demonstrated by significant increase in DCF staining. Although pre-treatment of cells with DXM did not affect BCNU cytotoxicity, BHA suppressed BCNU cytotoxicity by 25% (p &lt; 0.05). BCNU treatment also resulted in increased LAK cell-mediated elimination of HLS-EC (65%) compared to untreated cells (16%). Similar to its effect on direct BCNU cytotoxicity, BHA prevented BCNU treated HLS-EC killing by LAK cells by 25%. However, DXM did not influence LAK cell-mediated killing of BCNU-treated HLS-EC cells. These results suggest that both ROS generation and immune reaction against perturbed endothelial cells play a central role in BCNU-associated HLS-EC injury. In contrary to our findings on human alveolar epithelial cells, BHA ameliorated BCNU-induced direct and immune-mediated cytotoxicity on HLS-EC, but not DXM.
Conclusion: Reactive oxygen species scavenging may be useful in prevention of liver toxicity of BCNU.
Phase I Trial of CG53135-05 to Prevent Mucositis in Patients Undergoing High-Dose Chemotherapy (HDCT) and Autologous Peripheral Blood Stem Cell Transplantation (PBSCT).
Authors: Schuster Michael W. , Shore Tsiporah B. , Greenberg June , Jalilizeinali Bita , Possley Scott , Annino Vincent I. , Hahne William 
Published: April 2020

Abstract
Mucositis is a painful side effect of many transplant conditioning regimens used in HDC and PBSCT. This complication often requires treatment with potent narcotic analgesics and may even require intravenous patient-controlled analgesia (PCA) for adequate pain relief as well as total parenteral nutrition (TPN). Infections in this setting may also result from disruption of the mucosal barrier with subsequent migration of intestinal bacteria into the blood stream. Previous treatments with oral rinses and topical applications of soothing gels have largely been ineffective. Recently, a breakthrough class of drugs in the fibroblast growth factor (FGF) family holds great promise in more effectively ameliorating or even preventing oral mucositis (OM). We report on the results of a Phase I trial with CG53135-05, a novel investigational protein therapeutic (FGF-20) that promotes epithelial and mesenchymal cell proliferation in vitro and has demonstrated activity in animal models. 14 patients (ages 25–75) undergoing HDCT with PBSCT were treated with escalating doses of study drug, including 0.1 mg/kg, 0.2 mg/kg and 0.33 mg/kg (concentrations are determined by the UV method which are equivalent to 0.3, 0.6, and 1 mg/kg by the Bradford method previously used). Conditioning regimens used included melphalan (Mel 200), cyclophosphamide, carmustine and etoposide (CBV), carboplatin and thiotepa (CT), cyclophosphamide, etoposide and carmustine (CEC) and busulfan/cyclophosphamide (targeted BuCy). The primary objective of this phase I trial was to evaluate safety, tolerability and pharmacokinetics of CG53135-05. Patients (pts.) were also scored daily for presence of OM using both the WHO and OMAS (oral mucositis assessment scale) grading scales. 7/14 pts. in this study experienced no OM (including 2 Mel 200 patients), 5 pts. experienced only grade 1 OM. while 2 pts. (both treated with Mel 200) experienced grade 3 OM, and no pts. experienced grade 4 OM. 1 pt. experiencing grade 3 OM required TPN. Only 4 pts. experienced diarrhea that lasted more than 4 days and only 1 pt. had gut mucositis-associated (E. coli) bacteremia. The median day of engraftment (ANC&gt;500/uL) occurred on day 14 (range: day 11–19). Patients tolerated the study drug well with no significant side effects up to a dose of 0.33 mg/kg. At that dose, 2 pts. experienced an infusional reaction consisting of fevers, nausea, and mild hypotension. Pharmacokinetics were measured at all dose levels and will be presented. CG53135-05 is a member of a breakthtrough drug class (FGF family) that was well tolerated in autologous stem cell transplant patients at doses up to 0.33 mg/kg with apparent clinical effects in ameliorating or preventing OM - 12/14 pts, thus, avoided severe (grades 3–4) mucositis following HDCT. A larger Phase II clinical trial is planned.
t(8;21)-Positive AML with Mutant Kit Is a Novel Therapeutic Target of Imatinib Mesylate.
Authors: Asou Hiroya , Kyo Taiichi , Inaba Toshiya 
Published: April 2020

Abstract
Point mutations in the kinase domain of c-Kit are frequently associated with t(8;21)-acute myeloid leukemia (AML). In our study, eleven (26 %) of 43 patients had mutations: six Asp816Val, two Asp816Tyr, one Asp816Ala, one Asp816His, and one Asn822Lys. Here, we provide evidences that proliferation of leukemia cells expressing both the AML1-ETO chimera and a c-Kit mutation heavily depends on signals originating from mutated c-Kit, and this mutation is a possible therapeutic target for imatinib mesylate. We initially investigated effects of imatinib on the growth of the Kasumi-1 cell line, which harbors both t(8;21) and a c-Kit kinase domain mutation (Asn822Lys). Imatinib inhibited autophosphorylation of c-Kit at the standard concentration (0.1 μM), and induced cell cycle arrest and apoptosis in an even faster time course than Ph1-positive cell lines treated with this drug. By contrast, growth of SKNO-1, another t(8;21)-positive leukemia cells without c-Kit mutation was not affected by imatinib. To test whether imatinib is effective for Asp816 c-Kit mutants, we isolated t(8;21)-positive fresh leukemia cells from untreated patients. Numbers of cells with an Asp816 mutant from four patients after short-term cultures in the presence of imatinib (0.1 μM, 4 days) were 20–30 % of those in the absence of imatinib, while no significant difference was observed for cells isolated from four patients without c-Kit mutation. (Viability of fresh leukemia cells without imatinib was maintained over 80% during this short-term culture.) Moreover, autophosphorylation of mutated c-Kit in leukemia cells from one patient with an Asp816 mutant was inhibited by imatinib. Our results disagree with those of previous studies, which indicated that cells with c-Kit mutations in the kinase domain are resistant to imatinib in murine IL-3-dependent cells and human mast cell leukemia cells. This discrepancy could be explained by high expression levels of c-Kit mutants in IL-3-dependent cells by powerful ectopic promoters, since overexpression of Bcr-Abl kinase is one of the major causes of resistance to imatinib in the treatment of CML patients. In addition, drug metabolism may be different between human t(8;21)-positive leukemia cells and in murine IL-3-dependent cells or mast cell leukemia cells. Although t(8;21) in AML represents a favorable prognostic indicator for achievement of cure, a substantial number of these patients relapse and eventually die of their disease. Indeed, of five patients harboring both t(8;21) and c-Kit mutations who we identified and followed up for more than five years, four relapsed. Therefore, our results suggest that imatinib would be useful for eliminating minimal residual disease in these patients after achievement of complete remission.
Bone Marrow-Derived Stem Cells Reduce Radiation-Induced Damage to Salivary Glands.
Authors: Wierenga Pieter K. , Lombaert Isabelle , Visser Willy , Kampinga Harm H. , de Haan Gerald , Coppes Rob P. 
Published: April 2020

Abstract
The salivary glands are often included in the field of irradiation during radiotherapy of head and neck cancer. This can result in severe side-effects that reduces the quality of life of the patient and may even limit the treatment dose. Late damage to the salivary glands is mainly caused by exhaustion of the tissue specific stem cells. Post-irradiation replacement of salivary gland stem cells with donor stem cells may ameliorate radiation-induced complications. Bone marrow-derived stem cells (BMSC) have been shown to be multipotent and thereby able to engraft in many tissues after injury. In this study, we assessed the potential of BMSC to reduce irradiation-induced salivary gland damage. C57BL/6 mice were transplanted with bone marrow from eGFP transgenic animals. After two months the salivary glands of these chimeric mice were locally irradiated with 15 Gy. BMSC were mobilized 10, 30 and/or 60 days after irradiation by s.c. injection of rHu-PEG-G-CSF. Saliva secretion (μl/15 minutes) was measured up to 90 days after irradiation by pilocarpine induction. Hereafter, the glands were extirpated and examined for eGFP-expression. From every individual animal one parotid and one submandibular gland was used to prepare single cell suspensions in order to detect eGFP-positve cells by flow cytometry. The other parotid and submandibular glands were analyzed using confocal laser fluorescence scanning microscopy and light microscopy. G-CSF treatment yielded in an increase of saliva flow for all time points. The optimal time-point for mobilization, however, was 30 days after irradiation as is demonstrated by an improvement of salivary flow from 5 to 30% when compared to radiation alone. FACS analysis showed that up to 10% of the isolated cells were eGFP-positive. Microscopic analysis revealed a similar amount of positive cells and an improved morphology. Immuno-histochemistry using anti-SM-actin antibodies showed the close vicinity of actin and eGFP within the cells, demonstrating the occurence of BMSC derived myoepithelial cells in irradiated salivary glands. Furthermore, using cell-type specific antibodies, the meyoepethilial nature of the eGFP positive was revealed. In conclusion, the results show that BMSC home to severely damaged salivary glands after mobilization. Hence, BMSC mobilization could become a promising modality to ameliorate radiation-induced complications in salivary glands after radiotherapy.
Juxtaposition of the HPFH2 Enhancer Is Not Sufficient To Reactivate the g-Globin Gene in Adult Erythropoiesis in bYAC Transgenic Mice.
Authors: Xiang Ping , Ye Xin , Han Hemei , Stafford Mary , George Stamatoyannopoulos , Li Qiliang 
Published: April 2020

Abstract
Hereditary persistence of fetal hemoglobin (HPFH) is characterized by high-level expression of the g globin gene in adult patients. HPFH2 contains a genetic deletion of approximately 83.5 kb from the middle of intron 2 of the yb gene to the region 66 kb 3′ to the b gene. The deletion juxtaposes an enhancer located downstream to the 3′ breakpoint to the vicinity of the g gene. To understand the reactivation mechanism of this fetal stage specific globin, we introduced the deletion into a 213 kb b-YAC. Four transgenic lines bearing 1–3 intact copies of the YAC construct were established. Globin gene expression at different developmental stages was measured by RNase protection assays. In the HPFH2 transgenic mice the e and g genes were expressed at high levels similar to the wild type YAC mice in embryonic and fetal stages. Unexpectedly, the g gene was completely silenced in the adult mice. The failure of g gene reactivation by juxtaposition of the HPFH2 enhancer contradicts the results reported by the Forget lab (Mol. Cell. Biol.17:2076–2089, 1997) and the Strouboulis lab (Blood102:3412–3419, 2003). The discrepancy could be due to the differences between the constructs used in the different labs. The construct used in the Forget lab was a cosmid containing the mLCR linked a 13 kb Gg and Ag fragment. The cosmid used by the Strouboulis lab contained the 22 kb LCR and a 5 kb Ag fragment. The bYAC construct we used in this study contains the whole b globin locus and spanned from about 40 kb 5′ to the e gene to 100 kb 3′ to the b gene. The failure of g gene reactivation in the HPFH2 YAC mice suggests that additional elements, which are missing in the 213 kb bYAC construct, are involved in reactivation of the g gene in the HPFH patients. Based on data from literature (Bender et al., Mol. Cell5:387–393, 2000; Forrester et al., Genes & Dev.4:1637–1649, 1990) and our own studies (Ping et al., this meeting) we speculate that the 200 kb upstream region is the candidate for harboring this activity.
A Contigs Strategy of Southern Blot Hybridization: Screening for DNase I Hypersensitive Sites in the 200 Kb Region 5′ to the B-Globin Locus LCR.
Authors: Xiang Ping , Han Hemei , Fang Xiangdong , Stamatoyannopoulos George , Li Qiliang 
Published: April 2020

Abstract
Formation of DNase I hypersensitive sites is an indication of local disruption of chromatin conformation. It has been documented that HS sites are frequently associated with functional DNA sequences, such as, promoters, enhancers, and insulators. While Southern blot hybridization is the standard method to detect HS sites, this procedure is time-consuming and labor intensive. To improve the efficiency of HS detection through Southern blot hybridization, we designed a contigs strategy of Southern blot hybridization and test it in the 200 kb region 5′ to the LCR in the b-globin locus. Based on the human genome sequence we made physical maps of seven 6-bp-cut restriction enzymes in the 200 kb region. From the map we selected continuous contigs of 10 to 15 kb fragment; and designed hybridization probes for the 5′ and 3′ ends of each fragment (some probes can be used in two neighboring fragments). The screening was performed on erythroid (K562) and non-erythroid (Jurkat) cell lines. We found about 40 HS sites within the region. The major sites were either erythroid specific (for instance, HSs at −66 kb, −142 kb, and −236 kb, the cap site of the e-globin gene is +1), or non-erythroid specific (for instances, HSs at −111 kb, −164 kb, and −205 kb). These HS sites will be investigated for enhancer, promoter, and insulator function using transient and stable transfection studies. Due to the limited number of enzyme required and the fact that each blot could be used several times, this strategy can greatly expedite the screening process for presence of DNase I hypersensitive sites. Estimated efficiency of this screening approach is about 0.5 to1 Mb per person per year.
Association of Skin and Blood Dendritic Cells with Acute Graft Versus Host Disease after Human Haematopoetic Cell Transplantation.
Authors: Auffermann-Gretzinger Susanne , Eger Lars , Bornhaeuser Martin , Thiede Christian , Schaekel Knuth , Oelschlaegel Uta , Illmer Thomas , Schaich Markus , Ehninger Gerhard 
Published: April 2020

Abstract
Both number and origin of dendritic cells (DC) in the blood have been associated with acute graft-versus-host disease (aGvHD), graft-verus-leukemia effect, relapse and graft failure after human allogeneic haematopoietic cell transplantation (aHCT). Aim of this study was the systematic simultaneous investigation of skin and blood DC subtypes, the analysis of their host/recipient origin after HCT and the correlation of DC kinetics with treatment, outcome and incidence of aGVD and its treatment in 31 recipients of allogeneic HCT. Epidermal shave biopsies and peripheral blood samples were collected from patients pre and at defined time points following aHCT, DC from skin and blood isolated and examined by FACS and quantitative PCR for Short Tandem Repeat markers. A significant reduction in number of skin and blood DC subtypes was observed already before start of conditioning chemotherapy in patients compared to healthy volunteers. A majority of donor derived epidermal Langerhans Cells (median 95%, range 53–100%) and also dermal DC (median 94%, range 39–100%) was found in all examined patients even early after transplant independent from chemotherapy regimen, graft, occurrence of skin GvHD or time point after transplantation. Numbers of both skin DC subsets remained low for months post HCT. Reconstitution kinetics of CD 11c+ preDC1 and CD 123+ preDC2 blood DC were similar to each other and pre transplant numbers were reached again around day +112 post transplant. Recipients of grafts containing higher T cell numbers had lower preDC1 counts on day +28. PreDC2 reconstitution was negatively affected by steroid treatment. Patients with aGvHD tended to have more preDC1 on day +28 and lower numbers of both preDC subsets on day 56 post transplant. Residual host blood DC were rare at all time points (median &lt;3% at all investigated timepoints range 2–97%). The kinetics of DC reconstitution at different sites might become an important diagnostic tool to predict the occurrence of acute and chronic GvHD and to monitor the efficacy of prophylactic and therapeutic interventions.
Gadd45a and Gadd45b Protect Haematopoetic Cells from Ultraviolet Radiation Induced Apoptosis Via Distinct Signaling Pathways.
Authors: Gupta Mamta , Gupta Shiv K. , Hoffman Barbara , Liebermann Dan A. 
Published: April 2020

Abstract
Gadd45 expression, which is stress inducible, has been associated with growth arrest, but the exact role of gadd45 family genes in apoptosis still remains unclear. We have found that myeloid progenitor cells from gadd45a and gadd45b-deficient mice are more sensitive to ultra-violet radiation, VP-16 or daunorubicin induced apoptosis. indicating that gadd45a or gadd45b protect haematopoetic cells from DNA damaging agents. To determine, how gadd45a or gadd45b proteins exert their anti-apoptotic function, bone marrow cells from wild-type and gadd45a or gadd45b deficient mice were exposed to ultraviolet radiation (UV) and analyzed for expression of stress responsive kinases, including JNK and p38. It was observed that P38 and JNK were activated in wt bone marrow cells in response to UV but not in bone marrow cells defecient in gadd45a. Also, the transcription factor NF-kB was activated in wt bone marrow cells, but not in gadd45a−/− cells. The pharmacological inhibitor SB203580 specific for p38, increased apoptosis in reponse to UV, indicating that p38 is implicated in signaling myeloid cell survival. SB203580 was observed also to inhibit the expression of certain NF-kB target genes, including cIAP-1, c-IAP-2, bcl-2 and bcl-xl, in gadd45a+/+ cells but not in gadd45a deficient bone marrow cells. Taken together this data provides first evidence for the role gadd45a plays in the control of hematopoietic cell survival in response to UV, via modulation of P38 MAPK and NF-kB signaling pathways. Unlike in gadd45a−/− bone marrow cells, p38 activation appeared not to be impaired in gadd45b−/− cells, indicating that gadd45b is not involved in p38 activation in myeloid cells. However, UV induced JNK activation was sustained in gadd45b−/− myeloid cells compared to wt cells, indicating that gadd45b is a negative modulator of UV induced JNK signaling in myeloid cells. UV induced activation of MKK4 an upstream regulator of JNK also was impaired in gadd45b−/−. NF-kB was also found activated in wt cells, but not in gadd45b−/− cells. This data indicates that in bone marrow cells exposed to UV, NF-kB induced expression of Gadd45b plays a protective role against UV induced apoptosis via inhibition of MKK4 kinase which in turn results in suppression of JNK activity. Taken together this data provides evidence that Gadd45a and Gadd45b protect haematopoetic cells from genotoxic-stress induced apoptosis via distinct signaling pathways.