Automated analyzer evaluation of reticulocytes in bone marrow and peripheral blood of hematologic disorders.

The R-3000 reticulocyte analyzer uses flow cytometry with an argon laser as its light source. This analyzer stains residual RNA with auramine O to provide a reticulocyte maturation differential. Using the R-3000, we analyzed 119 samples of bone marrow (BM) and peripheral blood (PB) from 111 patients with hematologic disorders. Parameters were reticulocytes, immature reticulocyte fraction (IRF) percentage in BM and PB, BM/PB reticulocyte ratio, and BM/PB IRF ratio. Reticulocytes and IRF percentage in BM were significantly higher than in PB (p < 0.01). There was also a good correlation between reticulocyte percentages in BM and in PB (r = 0.81). Patients were classified into a normal group (without anemia) and an anemia group. Furthermore, the anemia group was classified into three groups: group 1: cases with hematopoietic dysfunction; group 2: cases in bone marrow recovery phase after chemotherapy and hematopoietic stem cell transplantation, and hematologic disorders with bone marrow accelerative phase, and group 3: cases with ineffective hematopoiesis (myelodysplastic syndrome). The mean reticulocyte percentage of the normal group was 2.3 +/- 1.1%, which was close to the normal value in BM. The BM/PB reticulocyte ratio of group 3 was statistically higher than that of groups 1 and 2. This indicates that group 3 had ineffective erythropoiesis and that the BM/PB ratio is a useful indicator for the diagnosis of myelodysplastic syndrome.

Evaluation of noninvasive hemoglobin monitoring for hematological disorders.

The Astrim is a noninvasive hemoglobin monitoring device that uses near-infrared radiation. We studied the usefulness of this device for patients with hematological disorders. We measured hemoglobin levels 309 times in 140 subjects, using an automated hematology analyzer (K-4500) and the Astrim. The coefficient of correlation between Hb levels with the K-4500 (K-Hb) and the Astrim (A-Hb) was r = 0.591 (P < 0.001). The coefficient of correlation between A-Hb and K-Hb from 174 specimens with anemia and 135 without anemia was r = 0.531 (P < 0.001) and r = 0.345 (P < 0.001), respectively. In the 309 specimens, the Astrim showed a diagnostic sensitivity and specificity for anemia of 78.3% and 69.0%, respectively. While the r-values, sensitivity, and specificity were passable, they appeared to be insufficient for an accurate evaluation. We believe this was caused by the measurement conditions (i.e., the finger selected for measurement, the finger position in relation to the CCD camera, and the finger temperature). On the other hand, the precision test results were good. Therefore, if careful attention is paid to measurement conditions, the noninvasive Astrim is clinically useful for continuous hemoglobin monitoring of hematological disorders and patients with hemorrhagic diseases, such as perioperative hemorrhage and gastrointestinal bleeding. However, it is difficult to determine Hb concentrations in one measurement using the Astrim because the r-values, sensitivity, and specificity are insufficient for accurate evaluations of in-patients, particularly patients with advanced diseases. Therefore, we believe it is necessary to improve the system so that A-Hb can be accurately determined in one measurement.

Automatic analysis of normal bone marrow blood cells using the XE-2100 automated hematology analyzer.

The bone marrow aspiration test conventionally has been performed by visual methods, using a light microscope, because automatic blood cell analyzers cannot adequately capture erythroblasts and immature granulocytes (IGs) (Tatsumi et al.: Osaka City Med J 1988;34:135-146; Tatsumi et al.: Am J Clin Pathol 1986;86:50-54). With the development of the XE-2100 automatic blood cell analyzer (Sysmex Corporation, Kobe, Japan) in 1999, the classification of erythroblasts and IGs by means of flow cytometry (Zini et al.: Infus Ther Transfus Med 2001;28:277-279; Briggs et al.: Sysmex J Int 1999;9:113-119) became possible. In the present study we classified cells in 65 bone marrow aspiration specimens by the microscopic method and with the XE2100, and compared the results. A good correlation was found in the nucleated red blood cell (NRBC), white blood cell (WBC), and total nucleated cell (TNC) counts; the myeloid/erythroid ratio (M/E ratio); neutrophils, lymphocytes, eosinophils, and IGs in the immature myeloid information (IMI) channel; and the total cell count. These items can all be analyzed in about 54 sec with the XE2100, which is faster than with the microscopic method. Therefore, analysis of bone marrow aspiration specimens with this apparatus appears to be very useful for clinical screening as well as laboratory testing.

[A case of amelanotic melanoma of the nasal cavity].

A 68-year-old male visited Hospital A for treatment of epistaxis, his chief complaint. He was told that he had an easily-bleeding tumor in the nasal cavity. Based of biopsy, a diagnosis of amelanotic melanoma was made. Operation was performed for removal of the tumor. About 8 months after discharge, he visited Hospital B with complaints of lumbar pain and epistaxis. After biopsy at Hospital B, malignant lymphoma (diffuse large cell) was diagnosed, and the patient was referred to our hospital. On bone marrow puncture and biopsy, tumor cell infiltration was observed. Flow cytometric surface marker analysis revealed that these tumor cells were negative for CD45. Results of HE staining of the nasal cavity tumor were insufficient for diagnosis, and staining by immunohistochemistry was necessary to confirm the diagnosis. On immunohistochemical staining of the nasal cavity tumor tissue and bone marrow biopsy tissue, LCA, L26 and UCHL-1 were negative, and S-100 and HMB-45 positive. Recurrence of amelanotic melanoma accompanied by bone marrow infiltration was therefore diagnosed. The incidence of amelanotic melanoma with primary lesions in the nasal cavity is low. However, in making the diagnosis of a nasal cavity lesion, the possibility of such a melanoma should be kept in mind. In many cases, it is difficult to diagnose amelanotic melanoma with HE staining alone, and immunohistochemistry must be used.