ALBERT

Description: Background/Aim: Hepcidin, a hepatic antimicrobial protein, is usually over-expressed in iron deficiency anemia. However, whether gastric Helicobacter pylori (H. pylori) infection effects a change in hepcidin level in iron deficiency anemia is still uncertain. We evaluated whether H. pylori eradication decreased hepcidin level and increased hemoglobin and ferritin levels in iron deficiency anemia. Method: From October 2006 to April 2007, nine females (mean age of 32.2 year-old) who were diagnosed as iron deficiency anemia without definite blood loss, were included in the study. All the subjects underwent upper gastrointestinal endoscopy and colonoscopy to diagnose gastric H. pylori infection and to exclude any source of gastrointestinal bleeding. Blood samplings for hemoglobin, hematocrit, iron (Fe), total iron binding capacity (TIBC), ferritin and hepcidin tests were taken just before H. pylori eradication and four weeks after H. pylori eradication and oral iron supplement therapy, respectively. Serum prohepcidin level was measured by Hepcidin Prohormone ELISA (Solid Phase Enzyme-Linked Immunosorbent Assay) kits. Statistical analysis was done by Wilcoxon signed rank test. Result: H. pylori eradication was successful in all the subjects who revealed negative in urea breath test. Hemoglobin, hematocrit, Fe, TIBC and ferritin levels improved in all the subjects after H. pylori eradication (Table 1). Mean serum hepcidin level was 224.8 ± 23.2 ng/ml initially, but decreased to 179.7 ± 40.8 ng/ml after H. pylori eradication therapy (p = 0.015). Conclusion: Our result provides evidence that hepcidin level decreases after successful H. pylori eradication with the improvement of iron deficiency anemia. A fall in serum hepcidin level resulting from successful H. pylori eradication reflects that hepcidin is an important mediator of iron absorption in iron deficiency anemia associated with gastric H. pylori infection. Table 1. Results of blood tests before and after Helicobacter pylori eradication Before H. pylori eradication After H. pylori eradication p-value* *Wicoxon signed ranks test Fe, iron; TIBC, total iron binding capacity Hemoglobin (g/dl, mean ± SD) 7.67 ± 2.01 10.76 ± 2.43 0.008 Hematocrit (%, mean ± SD) 26.36 ± 5.76 34.26 ± 5.76 0.008 Fe (ug/dl, mean ± SD) 11.78 ± 4.21 45.00 ± 54.39 0.038 TIBC (ug/dl, mean ± SD) 480.78 ± 51.39 427.00 ± 66.30 0.008 Ferritin (ng/ml, mean ± SD) 2.67 ± 0.99 7.39 ± 6.38 0.021 Hepcidin (ng/ml, mean ± SD) 224.80 ± 23.23 179.70 ± 40.84 0.015

Description: Intravenous (IV) busulfan has been developed to overcome variable absorption of oral busulfan and tested in several trials. We tested its pharmacological properties and tolerability in 16 Korean SCT patients. IV busulfan was administered at 0.8 mg/kg every 6 hours for a total of 16 doses (days -7 to -4), which was followed by cyclophosphamide administration at 60 mg/kg every 24 hours for 2 days (days -3 and -2). The median AUCinf values (at the 1st dose) and AUCss (at the steady state) were 1060.4 mM·min (range: 511.1– 1812.7) and 1092.5 mM·min (range: 539.7 – 1560.8), respectively. All patients had an AUCinf of 〈 1500 mM·min at the 1st dose, and 13 of the 16 (81.3 %) maintained AUCss levels between 800 and 1500mM·min. Overall, pharmacokinetics of IV busulfan in our SCT patients appeared comparable to those observed in other study. Thirteen of 16 patients achieved successful engraftments but 4 patients (25%) developed hepatic VOD (2 of which were fatal). Although there was no apparent correlation between AUC and hepatic VOD development, 3 out of 4 patients with VOD had advanced disease at the time of SCT. Notably, patients who had uncontrolled diseases (CML blast crisis, ALL 2nd relapse) succumbed to VOD and those who survived VOD had less advanced disease (CML CP, ALL in 2nd CR) at the time of SCT. In conclusion, pharmacokinetics of IV busulfan in our SCT patients appeared comparable to those observed in other studies, however, hepatic VOD was still a major morbidity, especially in patients with advanced disease, warranting consideration of other parameters in addition to pharmacokinetic monitoring of IV busulfan to decrease VOD in high risk patient populations. Hepatic veno-occlusive disease cases (n=4) sex/age disease state AUCinf AUCss Outcome Comment F/41 CML blast crisis 511.0 539.7 death unrelated donor F/49 CML chronic phase 739.4 991.0 recovered F/39 ALL 2nd CR 1472.1 1465.6 recovered M/17 ALL 2nd relapse 1302.7 1560.8 death pseudomonas sepsis

Description: Abstract 5101 Background Helicobacter pylori infection seems to subvert the human iron regulatory mechanism, and thus up-regulate hepcidin that results in unexplained iron deficiency anemia (IDA). We evaluated serum pro-hepcidin levels before and after H. pylori eradication in IDA patients to assess whether it plays a role in H. pylori-related IDA. Materials and Methods Subjects diagnosed as unexplained IDA underwent upper gastrointestinal endoscopy and colonoscopy to diagnose H. pylori infection and to exclude gastrointestinal bleeding. Blood sampling were done before H. pylori eradication and after a month. Serum pro-hepcidin level was measured by a commercialized enzyme-linked Immunosorbent assay kit. Results Initial serum pro-hepcidin levels were not different between 23 H. pylori-infected subjects (212.9 ± 88.2 ng/ml) and nine non-infected subjects (217.8 ± 56.2 ng/ml) (p=0.879). Serum pro-hepcidin level decreased after either dual oral iron replacement with H. pylori eradication (p=0.011) or H. pylori eradication without iron replacement (p=0.075). It also decreased after iron replacement in non-infected subjects (p=0.086). The reduction ratio of serum pro-hepcidin level after the treatment was not different between three groups (p=0.972). Conclusions Serum pro-hepcidin level decreases after either H. pylori eradication or oral iron administration with IDA improvement. Serum pro-hepcidin is related to the status of anemia rather than the presence of H. pylori itself. Disclosures No relevant conflicts of interest to declare.

Description: Background and Objectives Mesenchymal stem cells plays an important role in the hematopoietic stem cell engraftment condition with SDF-1 (CXCL12)-CXCR4 signaling and in their homing in various tissues. In this study, we evaluated that the regulation of homing efficiency for mesenchymal stem cells to support ex vivo expansion of hematopoietic stem cells from umbilical cord blood. Methods We investigated the expression of CXCR4 and Stromal-Derived Factor-1 (SDF-1) in cocultured mesenchymal stem cell with umbilical cord blood-derived CD34-positive cell, which stimulated with granulocyte macrophage-colony stimulating factor (GM-CSF) and stem cell factor (SCF) cytokine. Results In this study, we evaluated that coculturing of SDF-1+ mesenchymal stem cells with stimulated CD34+ cells significantly increased the expression of CD34, CD45, and CD19 for myeloid surface marker and intracellular CXCR4 within a few hours as compared with culturing of CD34-positive cells alone or with SDF-1− mesenchymal stem cells or untreated mesenchymal stem cells by Flow cytometre. In the result of stimulation for 48 hours with various cytokines in CD34-positive cells, CXCR4 gene and ERK-1,2 protein up-regulated, and increased in vitro migration capacity of cocultured SDF-1+ mesenchymal stem cell with CD34+ cells as examined by quantitative RT-PCR of human GAPDH. To enhance homing effect by mesenchymal stem cell, we maintained expanded mesenchymal stem cells for up to 5–10 passages with monitoring of the expression of various tissue surface antigens, such as skeletal muscle, neural, liver, and endothelial cells. SDF-1+ mesenchymal stem cells induced the homing of cellular products of stimulated cord blood-derived CD34-positive cells for 10 days. Moreover, the tranfected SDF-1+ cells with a green fluorescent protein gene using lentivirus maintained their capacities of protein release and homing in culture system. SDF-1− mesenchymal stem cells reduced CXCR4 expression in cocultured CD34-positive cells. Conclusions: These results demonstrate that the role of the SDF-1/CXCR4 axis is an important rold in the regulation of homing and engraftment of mesenchymal and hematopoietic stem cells. SDF-1+ mesenchymal stem cells have clinical potential to regulate homing and short-term engraftment for hematopoietic stem cell transplantation.

Description: Abstract 5282 BACKGROUND: The laboratory test with the highest sensitivity and specificity for the diagnosis of iron deficiency anemia (IDA) is serum ferritin. Chronic blood loss, one of the common causes of IDA in adults, is often associated with an occult gastrointestinal (GI) malignancy, especially in adult males and postmenopausal females. It is difficult to set the cutoff ferritin level for GI endoscopic evaluation in adult patients with anemia which is presupposed to result from GI malignancies. To answer this question, we conducted a retrospective study to find out the optimum values of serum ferritin and other hematologic indices in adult anemic patients to be referred for thorough GI endoscopic evaluation. METHODS: We reviewed retrospectively patients' medical records. The subject of study was adult anemic patients (n=544) at Konkuk University Medical Center who underwent upper and lower GI endoscopy to search for possible GI blood loss from August 2005 to August 2009. Patients were stratified into three groups according to the results of GI endoscopy: benign group vs. premalignant group vs. malignant group. RESULTS: Among a total of 544, benign, premalignant and malignant diseases were detected in 265, 220 and 59 patients, respectively. The prevalence of GI malignant diseases was 10.8% (59/544) for all patients, 13.9% (34/244) for male patients and 8.3% (25/300) for female patients. As compared to non-malignant groups (benign and premalignant), malignant group demonstrated statistically significant differences in terms of median values of age (56 vs. 66 vs. 70 years, P 〈 0.001), male gender (41.89 vs. 45 vs. 57.63%, P = 0.0367), MCV (86.7 vs. 88.2 vs. 78.6 fL, P = 0.0005), ferritin (69.39 vs. 108.5 vs. 21.7 ng/mL, P = 0.0002), Fe (35 vs. 38 vs. 13.5 μg/dL, P 〈 0.001), TIBC (279 vs. 259 vs. 320 μg/dL, P = 0.0024), and TIBC saturation (13.07 vs. 14.78 vs. 4.64%, P 〈 0.001). However, Hb, Hct and RDW did not show significant differences. By ROC curve analyses to find out optimum cut-off points of the serum ferritin and TIBC saturation which distinguish between non-malignant diseases and malignant diseases, the cut-off ferritin value of 44.33 ng/mL in male had a sensitivity of 72.73% and a specificity of 70.95% (AUC 0.705 and P = 0.0001). In contrast, ROC curve analyses were not useful for ferritin in female (AUC 0.609 and P =0.0787). The cut-off TIBC saturation value of 9.13% in male had a sensitivity of 73.33% and a specificity of 70.92% (AUC 0.750 and P = 0.0001). And the cut-off TIBC saturation value of 6.16% in female had a sensitivity of 69.57% and a specificity of 65.13% (AUC 0.643 and P = 0.0262). CONCLUSION: Our study proposes that adult male patients with anemia require thorough endoscopic evaluation to detect GI malignancy when their serum ferritin levels are ≤ 44 ng/mL or TIBC saturation values are ≤ 9%. For adult female anemic patients, only TIBC saturation values less than 6% may contribute to determining whether they require GI endoscopic evaluation. Disclosures: No relevant conflicts of interest to declare.