The first fluorescent sensor for boronic and boric acids with sensitivity at sub-micromolar concentrations--a cautionary tale.

The first reported fluorescent sensor for boronic and boric acids is actually not a sensor for boronic and boric acids but rather is a sensor for protons; the system is also not the first fluorescent sensor since Alizarin has been used as a fluorescent sensor for boric acids since 1936.

Modular fluorescence sensors for saccharides.

Modular and modular polymer supported fluorescence photoinduced electron transfer (PET) sensors 2 and 3 with two boronic acid receptor units, a pyren-1-yl fluorophore, and hexamethylene linker show selective saccharide binding in aqueous methanolic solution at pH 8.21.

Effect of recombinant human granulocyte colony-stimulating factor on combination therapy with aztreonam and clindamycin for infections in neutropenic patients with hematologic diseases.

The present multicenter study was performed to evaluate the effect of recombinant human granulocyte-colony stimulating factor (rhG-CSF) on combination therapy using aztreonam (AZT) and clindamycin (CLDM) to treat severe infection in neutropenic patients with hematologic diseases. Forty-three neutropenic patients with infections (rhG-CSF group) were treated with AZT (2 g) and CLDM (600 mg) 2-3 times daily as well as rhG-CSF (Lenograstim or Filgrastim: 2-5 mu/kg/day). The clinical efficacy of this regimen was compared to that obtained in 44 febrile neutropenic patients, with hematologic diseases, who received only AZT and CLDM in a previous study (historical control group). The overall efficacy rate was 69.8% (30/43) in the rhG-CSF group and 65.9% (29/44) in the historical control group. Although the neutrophil count was significantly increased and C-reactive protein tended to be lower in the rhG-CSF group, the daily maximum body temperature profiles of the 2 groups were nearly the same. These results suggest that rhG-CSF is of little benefit in the treatment of single infectious episodes in neutropenic patients, and that appropriate antibiotic therapy is more important.

Promotion of differentiation and proliferation of peripheral blood CD34+ cells in vitro by G-CSF.

OBJECTIVE: To observe the effect of granulocyte-colony stimulating factor (G-CSF) on differentiation and proliferation of CD34+ cells from peripheral blood in presence of recombinant hematopoietic growth factor (HGF). METHODS: Peripheral blood mononuclear cells mobilized by G-CSF were obtained from patients suffering from carcinoma, preparing for autologous bone marrow transplantation. CD34+ cells were isolated by derivatized polystyrene tissue culture flask which had covalently immobilized soybean agglutinin lectin and was coated with anti-CD34 antibody; and this kind of cells were incubated in liquid culture medium for up to 28 days under the stimulation of combination of growth factors, i.e., stem cell factor (SCF) and interleukin-3 (IL-3) with or without G-CSF. The changes of nucleated cells, colony forming unit-granulocyte and monocyte (CFU-GM), burst forming unit-erythrocyte (BFU-E), colony forming unit-megakaryocyte (CFU-MK), and myeloid-associated markers were evaluated. RESULTS: An increase of nucleated cells (mean 640-fold increase) occurred during culture CFU-GM production is parallel to the nucleated cell production until the 11th day (mean 82-fold increase) in combination of 3 HGF, i.e., G-CSF, IL-3 and SCF. A large number of cells expressing late myeloid markers appeared on the 11th day in suspension culture of CD34+ cells. CONCLUSION: G-CSF was found to synergize with IL-3 and SCF in inducing rapid proliferation of purified CD34+ cells and differentiation to multiple myeloid lineages. The stroma-free, cytokine-driven culture system could achieve a degree of amplification of colony forming cells, suggesting the feasibility of culture of hematopoietic progenitor cells in vitro as an adjunct to hematopoietic stem cell transplantation.

Nucleotide sequence of a human genomic DNA fragment containing the PCNA pseudogene and its localization on chromosome 4.

A one kb human genomic DNA fragment, containing a processed pseudogene of a proliferating cell nuclear antigen (PCNA/DNA polymerase delta auxiliary protein), was isolated and sequenced. The PCNA pseudogene consisted of the 3' half of exon 4 and the 5' half of exon 5 of the PCNA gene, and shared 84% nucleotide homology with the human PCNA cDNA. The PCNA pseudogene was localized on human chromosome 4, based on data obtained from a panel of human-mouse hybrid cell lines.

[Pulmonary functions in patients with progressive systemic sclerosis].

We studied the correlations of pulmonary function with clinical symptoms or autoantibody profiles in patients with progressive systemic sclerosis (PSS). Percent vital capacity (% VC) was low in 40% of PSS patients and percent diffusing capacity of carbon oxide (% DLCO) was decreased in 76% of the patients. Eighty-five percent of the patients had abnormal pulmonary functions. The % DLCO was significantly low in male patients compared with female patients. The arterial partial pressure of oxygen (PO2) in patients with esophageal hypomotility was significantly lower than those without esophageal hypomotility. PSS with Sjögren's syndrome (SS) showed a significant decrease of the % VC (p < 0.05) and PO2 (p < 0.05), and of lung fibrosis was frequent in the patients compared with those without SS. Pulmonary fibrosis was recognized in 63% of PSS patients and was more frequent in the patients with anti-Scl-70 antibody than those without the antibody. The annual decrease of VC was 62 ml in PSS patients, and that of DLco was 0.12 ml/min/mmHg. The annual decrease of VC was more prominent in the PSS patients with severe pulmonary fibrosis or anti-Scl-70 antibody. The decrease of VC, however, was mild in the patients with anti-RNP antibody compared with those without the antibody.

Combinations of HLA-DPB1 and HLA-DQB1 alleles determine susceptibility to early-onset myasthenia gravis in Japan.

HLA class II alleles in the DQA1, DQB1, DRB1, and DPB1 genes were investigated in Japanese patients with myasthenia gravis (MG) by digestion of polymerase chain reaction amplified DNAs with allele specific restriction endonucleases (PCR-RFLP). A significantly higher frequency of DQB1*03, which includes *0301, *0302, *0303 and determines the serological DQ3 specificity, was observed in female patients less than 30 years in age at onset of disease compared with healthy controls (90.5 vs. 53.2%). This study also confirms the high incidence of DPB1*0201 in early-onset female patients compared to the controls (85.7 vs. 40.3%). Moreover, 81.0% of the early onset female patients were found to carry both DQB1*03 and DPB1*0201, compared to 17.7% of the controls. Since DQB1*03 and DPB1*0201 are not in linkage disequilibrium, both these alleles are supposed to be synergistically involved in disease development in early-onset female MG. In contrast, no obvious association of HLA-DQA1, DQB1, DRB1 and DPB1 alleles with either late-onset patients or patients with thymoma was observed. Clearly, the genetic background of Japanese females with early onset MG is different from that of other patients with MG.

Flow cytometric analysis of cell-surface antigen expressions on acute myeloid leukemia cell populations according to their cell-size.

Acute myeloid leukemia (AML) cells which expanded from a single leukemic cell show certain degrees of morphological and biological heterogeneity. In the present study, we determined cell-surface antigen expressions (CD13, 33, 34 and 38, and HLA-DR) on AML cells based on their cell-size (large vs small cells) by flow cytometry. We found that the cell-surface antigens were more strongly expressed on the large leukemic cells than the small cells, regardless of FAB subtypes. Furthermore, our preliminary study demonstrated that AML patients who showed a relatively small difference in antigen expression between large and small leukemic cells had longer remission durations and survival periods, compared with those with a more prominent difference in antigen expression. Thus, the heterogeneity of AML cells determined by the combination of cell-surface antigen expressions and cell-size may be associated with clinically important biological behaviors.

[Renal and cerebral infarctions in a patient with systemic lupus erythematosus without antiphospholipid antibodies].

Renal artery infarction is a very rare complication in patients with systemic lupus erythematosus (SLE), even in patients with antiphospholipid syndrome which often causes thromboembolism: Renal infarctions have only been reported in 4 SLE patients with antiphospholipid antibodies (aPL). Here we report a case of SLE without aPL who accompanied by renal and cerebral infarctions. A 42-year old Japanese woman with 8 year history of SLE manifested by arthralgia, central nervous system symptoms, positive-antinuclear and anti-DNA antibodies was admitted to our hospital for the treatment of progressive lupus nephritis. Physical examinations revealed hypertension (130-160/80-110 mmHg) without pitting pretibial edema. Laboratory evaluations showed proteinuria (3.7 g/day), normal serum creatinine level (0.9 mg/dl), low serum albumin level (2.3 g/dl) and high cholesterol level (317 mg/dl). Old cerebral infarctions were recognized by magnetic resonance imaging. However, hematological and immunological studies revealed that this case has neither a prolonged activated partial thromboplastin time, lupus anticoagulant nor anticardiolipin antibodies. Prednisolone was increased from 30 mg/every other day to 30 mg/day, and oral azathioprine, 50 mg/day, was started for the treatment of lupus nephritis. On the 11th day, she suddenly complained severe abdominal pain, which gradually localized on the right side. Computed tomography of the kidney suggested right renal infarctions, and arteriography of right renal artery confirmed both an obstruction of the ventral branch and a narrowing of the dorsal branch of right renal artery. No intra-cardiac thrombus was demonstrated by echocardiography. Following to the treatment with fibrinolytic agent and anticoagulant, her symptoms have improved.(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of c-myc, c-Ki-ras and c-Ha-ras oncogene products in peripheral blood mononuclear cells from patients with myasthenia gravis.

The oncogene products of c-myc, c-Ki-ras and c-Ha-ras and the subsets of natural killer cells in the peripheral blood mononuclear cells (PBMC) from 39 patients with myasthenia gravis (MG) and 40 healthy individuals were studied by immunocytochemical procedures. The MG patients showed an increased expression of c-myc, c-Ki-ras and c-Ha-ras oncoproteins in both lymphocytes and monocytes compared to those of healthy individuals (P < 0.01, P < 0.005 and P < 0.001, respectively). On the contrary, the number of Leu-7-Leu-11c+ cells of MG patients was less than that of healthy controls (P < 0.05). The oncogene expressions in PBMC of MG patients correlated positively with white blood cell counts, total natural killer cell counts (NK), Leu-7+Leu-11c+ of NK cell subset, gamma-globulin, erythrocyte sedimentation rate and serum immunoglobulins; the oncogene expressions were negatively correlated with the Leu-7+Leu-11c-NK subset. Thus, the increased expressions of c-myc, c-Ki-ras and c-Ha-ras oncoproteins accompanying the decreased Leu-7-Leu-11c+NK cell in MG PBMC are likely responsible for the acceleration of the clinical manifestations in MG patients.

[Estimation of megakaryothrombopoiesis by mean platelet volume in leukemia patients during the course of chemotherapy and patients undergoing bone marrow transplantation].

The mean platelet volume (MPV) was measured in 14 patients with acute leukemia during the course of chemotherapy for remission induction and 21 patients undergoing bone marrow transplantation, in order to assess its usefulness as an indicator for megakaryothrombopoiesis at bone marrow suppression and recovery. In patients with acute leukemia, the MPV fell from 9.7 +/- 0.8 fl to 9.2 +/- 0.4 fl during the myelosuppression, and then it rose transiently to 10.2 +/- 0.5 fl and fell to 9.3 +/- 0.5 fl during recovery of platelet count. Similarly, in patients undergoing bone marrow transplantation, the MPV elevation was followed by the recovery of platelet count. Thus, when engraftment was defined as this MPV elevation, engraftment was confirmed significantly earlier by MPV (19.9 +/- 5.8 days) than by platelet count (24.8 +/- 6.7 days). MPV should be a useful indicator for engraftment or recovery from marrow aplasia in cases of acute leukemia and those of bone marrow transplantation.

Pulmonary fibrosis with megakaryocytoid cell infiltration in accelerated phase of chronic myelogenous leukaemia.

We report a patient in the accelerated phase of Philadelphia-chromosome-positive chronic myelogenous leukaemia who developed fibrosis in lungs, spleen and bone marrow. In the lungs, fibrosis was demonstrated in the alveolar septa which had been infiltrated by giant, bizarre-shaped cells resembling the megakaryocytes in the bone marrow. The relationship between the fibrosis and megakaryocytoid cell infiltration in the lungs is discussed.

[Soluble IL-2 receptor levels in rheumatic diseases].

We measured soluble interleukin-2 receptors (sIL-2R) in plasma and/or joint fluid (JF) from patients with rheumatoid arthritis (RA: 16 patients), Sjögren's syndrome (SS: 24 patients) and Behçet's disease (BD: 23 patients). Plasma sIL-2R levels were significantly higher in RA patients (956 +/- 994 U/ml, p < 0.01) and SS patients (1025 +/- 1112 U/ml, p < 0.01) than 20 healthy subjects (338 +/- 159 U/ml), but not significantly increased in BD patients (408 +/- 329 U/ml). sIL-2R levels higher than the mean of healthy subjects +2 standard deviations were observed in 3 BD patients (13.0%), 7 RA patients (43.8%) and 10 SS patients (41.7%). sIL-2R levels positively correlated with erythrocyte sedimentation rates and C-reactive protein levels in RA patients, or IgM levels in SS. There was no significant correlation between plasma sIL-2R levels and disease activity, clinical symptoms, or other laboratory parameters in BD patients. However, sIL-2R concentrations in the JF from 4 BD patients (1644 +/- 198 U/ml: p < 0.05) or 16 RA patients (3946 +/- 5424 U/ml: p < 0.01) were significantly higher than corresponding plasma sIL-2R concentrations. Above results indicate that lymphocytes are being activated in patients with RA or SS and in the inflammatory lesions of BD patients, while the whole immunologic activation are relatively lower in BD than RA and SS.

[Refractory acute myelogenous leukemia successfully treated with a continuous infusion chemotherapy of low dose aclarubicin and cytosine arabinoside: a case report].

A 44-year-old female with refractory acute myelogeneous leukemia had experienced an early relapse after a third complete remission (CR) with combination chemotherapy (BHAC-AMP) using daily one-shot infusion of 20 mg aclarubicin (ACR). Further treatments including intermediate dose of cytosine arabinoside (Ara-C) were not effective. After treatment with a continuous administration of low dose ACR (9 mg/day) and Ara-C (21 mg/day) for 14 days she entered a fourth CR. The mechanism of this treatment relates to both cell-killing and differentiation effects on leukemic cells.

HLA-DPB1 allele associates with early-onset myasthenia gravis in Japan.

We investigated the HLA-DPB1 allele in 43 unrelated Japanese patients with myasthenia gravis (MG) using digestion of polymerase chain reaction (PCR)-amplified DNA with allele-specific restriction endonucleases (PCR-RFLP method). We found a higher frequency of the DPB1*0201 allele in female patients whose ages at onset were less than 30 years (83.3%) than in controls (35.6%). The study also included serologic typing of HLA-A, -B, -C, and -DR antigens in 72 patients with MG, and confirmed previous results demonstrating a strong association of HLA-DR53 with early onset of MG in females. These results indicate that both the DPB1*0201 allele and DR53 play key roles in the disease process of MG in early-onset females, and that the genetic background of Japanese females with early-onset MG is different from that of other patients with MG.

[Hematologic response to low dose natural interferon-alpha in a case of CML with myelo-megakaryoblastic crisis].

A 59-year-old female with splenomegaly was admitted in November, 1989. Her WBC was 7,900/microliters with 51% myeloblasts and 10% megakaryoblasts. Analysis of the surface markers showed that 56.5% were CD13 positive and 66.8% carried platelet GpIIb/IIIa. The Ph1 chromosome was 100% positive. VPM therapy was started but proved ineffective, as was subsequent MCNU therapy. The patient was given intramuscular human lymphoblastoid interferon-alpha (1.2 million IU daily) for more than 20 months. She had improved to the accelerated phase after INF-alpha therapy for 13 months. Thus there appears to be a relationship between INF-alpha and myelo-megakaryoblastic crisis.

Phase IV group study of natural human interferon alpha (HLBI) on chronic myelogeneous leukaemia.

Under the control of Kanagawa CML/HLBI phase IV study group in Japan, 18 cases out of registered 30 cases of chronic myelogeneous leukaemia consisting of 17 chronic phase and 1 accelerated phase, during July, 1991 to January, 1992, were analyzed for their hematological responses and cytogentic responses preliminarily. Hematological response rate (PR + CR) was 83.3% including 50.0% of CR, as judged by Kimura's criteria after treatment with HLBI alone (16 cases) or/and with other chemotherapy (2 cases). The dosage and duration of HLBI therapy required to get into the complete remission ranged from 212 to 1272 millions IU and between 6 to 42 weeks (mean value was 20 weeks), respectively. The clonally proliferated leukocytes and decreased physiological hematopoiesis started to recover from 2 to 4 weeks and reached their normal ranges from 16 weeks after 6 millions IU of HLBI were administered every day. In the 4 cases examined, 3 cases showed minimal cytogenetic responses and a case showed no cytogenetic response. Slight and temporary adverse effects were observed in 15 out of 18 cases (83.3%) including fever, general malaise, appetite loss, eruption, diarrhea, glossitis, hypogustation, weight loss and local muscle pain.