Human leukocyte antigen-DR expression on flow cytometry and tumor-associated macrophages in diffuse large B-cell lymphoma treated by rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone therapy: retrospective cohort study.

Loss of human leukocyte antigen (HLA)-DR expression may be related to a poor prognosis of diffuse large B-cell lymphoma (DLBCL), and tumor-associated macrophages (TAMs) may influence tumor progression. We retrospectively reviewed 36 patients with newly diagnosed DLBCL who received R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone) therapy at Kanagawa Cancer Center in Japan from 2004 to 2010. HLA-DR expression by lymphoma cells was evaluated using flow cytometry, and TAMs in lymphoma tissue were detected by immunohistochemistry for CD68 as a marker of macrophages and CD163 as a marker of M2 TAMs. Three-year overall survival was, respectively, 100% versus 69.6% in the HLA-DR "bright" and "not bright" groups (p = 0.012). Patients from the HLA-DR "not bright" group with strong CD163 expression had a much worse prognosis than other patients. The HLA-DR status shown by flow cytometry can be used to predict the prognosis of patients with DLBCL receiving R-CHOP therapy and prognostic accuracy can be increased by also assessing TAMs.

[Immunophenotypic analysis of hematogones in patients with hematological malignancies].

We studied immunophenotypic analysis of hematogones by flow cytometry. A total of 102 specimens from 93 patients with acute leukemia (52 specimens), myelodysplastic syndromes (4), or malignant lymphoma (46) were analyzed between April and August, 2011. Hematogones were detected in 55 specimens and highly identified in patients with acute myeloid leukemia in remission and B cell lymphoma. Stage 1 (CD34(+)CD20(-)) and stage 2/3 (CD34(-)CD20(+)) were detected in 9.9% and 52.7%, respectively. In addition, the intermediate type (CD34(+)CD20(+)) was identified in 37.4%. All specimens of stage 3 in bright CD45 expression were positive for CD5 and included CD5(+)CD23(-)CD11c(-), 11.1%, CD5(+)CD23(+)CD11c(-), 85.2%, and CD5(+)CD23(+)CD11c(+), 3.7%. These findings suggest that hematogones with unreported immunophenotypes may exist and the appearance of hematogones in hematologic malignancies may be relatively frequent.

The impact of the dose of natural killer cells in the graft on severe acute graft-versus-host disease after unrelated bone marrow transplantation.

The impact of lymphocyte subpopulations on the outcome of bone marrow transplantation (BMT) remains uncertain. We investigated the relationship between the lymphocyte subpopulations of bone marrow grafts and the outcome of BMT. A total of 121 patients who underwent BMT at Kanagawa Cancer Center between 2000 and 2009 were analyzed. Grade III-IV acute graft-versus-host disease (GVHD) occurred in 35.9% of patients who received unrelated BMT with a CD56 cell dose ≤2.80×10(6)/kg versus only 9.7% of patients with a CD56 cell dose >2.80×10(6)/kg (P=0.017). In patients receiving related BMT, the cumulative incidence of grade III-IV acute GVHD did not differ significantly in relation to the CD56 cell dose. On multivariate analysis, older donor age (hazard ratio (HR): 1.09, 95% confidence interval (CI): 1.03-1.15, P=0.004) and a high dose of CD56 cells (>2.80×10(6)/kg) (HR: 0.15, 95%CI: 0.03-0.92, P=0.040) were significant determinants of grade III-IV acute GVHD after unrelated BMT. None of the lymphocyte subpopulations had a significant impact on the outcome of transplantation, including the rate of neutrophil engraftment, relapse, relapse-free mortality, and overall survival. Our findings suggest that a high natural killer cell dose prevents severe acute GVHD after unrelated BMT, while sparing the graft-versus-leukemia effect.

Production of starch with antioxidative activity by baking starch with organic acids.

A starch ingredient with antioxidative activity, as measured by the DPPH method, was produced by baking corn starch with an organic acid; it has been named ANOX sugar (antioxidative sugar). The baking temperature and time were fixed at 170 °C and 60 min, and the organic acid used was selected from preliminary trials of various kinds of acid. The phytic acid ANOX sugar preparation showed the highest antioxidative activity, but the color of the preparation was almost black; we therefore selected L-tartaric acid which had the second highest antioxidative activity. The antioxidative activity of the L-tartaric acid ANOX sugar preparation was stable against temperature, light, and enzyme treatments (α-amylase and glucoamylase). However, the activity was not stable against variations in water content and pH value. The antioxidative activity of ANOX sugar was stabilized by treating with boiled water or nitrogen gas, or by pH adjustment.

The presence of mature granulocytes/monocytes derived from leukemic cells in MLL-associated leukemia.

We observed the mature granulocytes/monocytes derived from leukemic cells in patients with acute myeloid leukemia who present mixed lineage leukemia gene (MLL). Morphologic observation and fluorescence in situ hybridization analysis (FISH) for chromosome 11q23 abnormality were studied, and a multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was done to identify the fusion partners with MLL. The bone marrow cells with FISH signals of MLL showed the cell differentiation of the myeloid and/or monocytic lineages in 4 of 6 AML patients. MLL partner genes were AF6, AF9, ELL, and ENL, respectively. There was no correlation between the fusion partner and the appearance of mature cells derived from MLL clones. RT-PCR showed the fusion between MLL exon 9 or 10 and the partner genes in mature granulocytes/monocytes. These findings suggest that subgroup of leukemia cells with MLL rearrangement has the differentiation potential of leukemic cells and mature granulocytes/monocytes derived from MLL clones may be biologically different from normal mature cells.

Four-color flow cytometric analysis of myeloma plasma cells.

We monitored the behavior of residual myeloma plasma cells in patients with multiple myeloma after high-dose therapy and autologous or allogeneic transplantation using 3 methods of a flow cytometric technique using 4-color staining, immunofixation, and polymerase chain reaction approaches. We analyzed 17 cases by a relatively simple flow cytometric technique using CD38/CD45/CD19/CD56. Detectable myeloma plasma cells were found in 5 patients at diagnosis and 9 patients after treatment. Of 14 cases, 9 (64%) had CD19-CD56+ myeloma plasma cells, and 5 (36%) of 14 had CD19-CD56- myeloma plasma cells. When 37 bone marrow samples that had less than 5% myeloma plasma cells were assessed, myeloma plasma cells were detected in all 20 immunofixation-positive cases and 3 of 17 immunofixation-negative cases (P = .002). All 4 polymerase chain reaction-negative samples characterized as immunofixation-negative contained no detectable myeloma plasma cells. Flow cytometry can provide effective information to detect low levels of myeloma plasma cells.

[Cord blood transplantation after successful treatment of brain abscess caused by Bacillus cereus in a patient with acute myeloid leukemia].

Central nervous system infection caused by Bacillus cereus is a rare condition, which often progresses rapidly and is fatal in immunocompromised patients. A 54-year-old woman with acute myelogenous leukemia fell into a coma with high fever during severe neutropenia while undergoing chemotherapy. A blood culture demonstrated the presence of B. cereus and magnetic resonance imaging showed multiple abnormal lesions in her brain. The patient was treated with meropenem and vancomycin, and recovered from the coma in a week. Antibiotic therapy was administered for seven weeks, and then she underwent cord blood transplantation for refractory acute myelogenous leukemia with successful engraftment without exacerbation of the brain abscess. This case demonstrates that brain abscess caused by B. cereus can be treated without surgical treatment.

[Fatal acute visceral disseminated varicella-zoster virus infection in a patient with chronic graft-versus-host disease].

A 34-year-old woman was admitted for chronic graft-versus-host disease ten months after an unrelated bone marrow transplantation. Cytomegalovirus (CMV) antigenemia on day 5 of hospitalization was negative. Thrombocytopenia occurred on day 9. Laboratory findings revealed severe liver dysfunction on day 13. On day 14, the patient developed interstitial pneumonia and disseminated intravascular coagulation, and died from progressive respiratory failure. Multinucleated giant cells were found in the lung, liver, spleen, esophagus, pancreas and intestine obtained at autopsy. Varicella-zoster virus (VZV) was detected in the blood using the polymerase chain reaction (PCR) technique. CMV, herpes virus type 6 and Epstein-Barr virus were detected in the liver and lung with the PCR technique. We concluded visceral VZV infection was the main cause of her death because of her aggressive clinical course and the histology at autopsy. In this case, chronic GVHD and its immunosuppressive treatment resulted in her fatal VZV reactivation.

[Clinical significance of detecting strong bcl-2 protein expression with flow cytometry in follicular lymphoma].

The expression of cell surface antigens and bcl-2 protein was examined with flow cytometry in 30 lymphoma samples including 28 lymph nodes and others, from January, 2001 to May, 2003. Cases studied included the following: follicular lymphoma (FL), 11; diffuse large B-cell lymphoma (DLBCL), 12; diffuse small B-cell lymphoma, 1; mantle cell lymphoma, 1; MALT lymphoma, 1; precursor-B lymphoblastic lymphoma, 1; T-lymphoblastic lymphoma, 1; blastic NK-cell lymphoma, 1; and unknown, 1. In addition, cytogenetic analysis, fluorescence in situ hybridization (FISH), and a real-time polymerase chain reaction assay were performed. Strong bcl-2 expression was observed in 91% of FL, and 17% of DLBCL. On the other hand, strong bcl-2 expression was found in all 9 cases of FL with t(14;18), all two cases of DLBCL with t(14;18), one unknown case with t(14;18), and one FL case with t(11;14). Our results support that strong bcl-2 expression as demonstrated with flow cytometry is correlated with FL (p<0.0001), and lymphomas with the t(14;18) (p<0.0001).

Inter-relationship between electrophoretic characteristics of pseudocereal and cereal proteins and their microscopic structure for possible substitution based on nutritional evaluation.

Amaranth, soybean and maize were screened for proteins and their nutritional value. Isopropanol-soluble protein and buffer-soluble protein fractions were extracted from seeds and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The comparison of the identity and differences between investigated plants was carried out by the obtained SDS-PAGE electrophoretic patterns, and their microstructure was determined by scanning electron microscopy. Electrophoretic patterns of extracted proteins have shown that the main protein subunits were concentrated between 10 and 50 kDa. Variations were found in major fractions and minor bands as well as in the fine structure. The microstructure of pseudocereal and cereal protein fractions was inter-related with the results obtained by their electrophoretic separation. Pseudocereal amaranth can be used as a nutritive substitute of cereal maize in functional foods.