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: 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: 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.

Description: Background: The recent discovery of a single point mutation in the JAK2 gene (V617F) is a major advance in our understanding of the pathogenesis of BCR/ABL-negative chronic myeloproliferative disorders (CMPD). The frequency of the JAK2 V617F mutation in CMPD differs according to methods and research groups. We investigated the frequency of JAK2 mutations in Korean CMPD patients and demonstrated their usefulness as a new molecular marker for treatment response and disease progression in BCR/ABL-negative CMPD using quantitative real time PCR and pyrosequencing. Methods: Seventy-eight patients with BCR/ABL-negative CMPD comprising 42 cases of essential thrombocythemia (ET), 26 of polycythemia vera (PV), 7 with idiopathic myelofibrosis (MF), and 3 unclassifiable (UC) CMPD were enrolled in this study. A 364-bp PCR product containing the JAK2 V617F mutation was sequenced in both directions from total bone marrow cells. Restriction enzyme-based assessment with BsaXI was also used to search for JAK2 mutations. A quantitative real-time PCR-based allelic discrimination assay and pyrosequencing (Pyrosequencer PSQ96) were used to quantify the JAK2 V617F mutation status. Results: A single JAK2 V617F point mutation was identified in 20 (77%) of 26 patients with PV, 12 (26%) of 42 with ET, 7 (100%) of 7 with MF, and 2 (67%) of 3 with UC. The V617F mutation was present in over half of Korean patients with BCR/ABL-negative CMPD, giving an overall frequency of 53%. The proportion of mutant alleles ranged from 36 to 100% in the real-time PCR and pyrosequencing analysis. Patients with MF had a higher percentage of JAK2 mutant alleles than patients with ET (MF〉PV〉ET). PV patients (n=8) with a good response (phlebotomy only) had a relatively low proportion (58.9±20.1%) of the JAK2 mutant, while patients (n=9) showing a poor response (additional chemotherapy) initially had a higher proportion (76.6±19.0%; p=0.04185). Conclusion: The frequency of the JAK2 V617F mutation in Korean patients with PV was lower than that in reports from Western countries, although the overall frequency in Korean patients with BCR/ABL-negative CMPD was similar to previous reports. The results also suggest that the JAK2 mutation (quantification of JAK2 mutant alleles) can be used as a new marker for treatment response and disease progression in BCR/ABL-negative CMPD using quantitative real-time PCR or pyrosequencing.