Altitudinal flower size variation correlates with local pollinator size in a bumblebee-pollinated herb, Prunella vulgaris L. (Lamiaceae).

The influence of locally different species interactions on trait evolution is a focus of recent evolutionary studies. However, few studies have demonstrated that geographically different pollinator-mediated selection influences geographic variation in floral traits, especially across a narrow geographic range. Here, we hypothesized that floral size variation in the Japanese herb Prunella vulgaris L. (Lamiaceae) is affected by geographically different pollinator sizes reflecting different pollinator assemblages. To evaluate this hypothesis, we posed two questions. (1) Is there a positive correlation between floral length and the proboscis length of pollinators (bumblebees) across altitude in a mountain range? (2) Does the flower-pollinator size match influence female and male plant fitness? We found geographic variation in the assemblage of pollinators of P. vulgaris along an altitudinal gradient, and, as a consequence, the mean pollinator proboscis length also changed altitudinally. The floral corolla length of P. vulgaris also varied along an altitudinal gradient, and this variation strongly correlated with the local pollinator size but did not correlate with altitude itself. Furthermore, we found that the size match between the floral corolla length and bee proboscis length affected female and male plant fitness and the optimal size match (associated with peak fitness) was similar for the female and male fitness. Collectively, these results suggest that pollinator-mediated selection influences spatial variation in the size of P. vulgaris flowers at a fine spatial scale.

Completion task to uncover consumer's perception: a case study using distinct types of hen's eggs.

The choice and consumption of eggs are made considering a consumers' multidimensional perception, and their understanding becomes essential to the production targeting and the products' success in the market. In this context, this work aimed to verify the consumers' perception about the distinct types of hens' eggs, using a projective technique of completion task combined with presentation of images. A hundred consumers (n = 100) evaluated the main factors, both positive and negative, involved at the purchase time of eggs besides estimating their price. Between the positive factors that guide the eggs' consumption and purchase, the category with highest mention of terms was "Health," whereas negatively it was highlighted the category "Price." Concerning the perception of price, the results showed that the factory farm white eggs' value was the one that least differed from the average market price, possibly due to the nearness and familiarity with this variety. The methodology of completion task combined with presentation of images proved as being a practical and efficient tool to capture the consumers' perception of eggs, capable of providing valuable information to the ones involved in the production chain and commercialization of these products.

Delay and attrition before treatment initiation among MDR-TB patients in five districts of Gujarat, India.

Setting: Gujarat, a state in west India. Background: Although treatment initiation has been improving among patients diagnosed with multidrug-resistant tuberculosis (MDR-TB) in programme settings, it has still not reached 100%. Objectives: To determine pre-treatment attrition (not initiated on treatment within 6 months of diagnosis), delay in treatment initiation (>7 days from diagnosis) and associated factors among MDR-TB patients diagnosed in 2014 in five selected districts served by two genotypic drug susceptibility testing (DST) facilities and a drug-resistant TB centre in Gujarat. Design: This was a retrospective cohort study involving record review. Results: Among 257 MDR-TB patients, pre-treatment attrition was seen in 20 (8%, 95%CI 5-12). Patients with 'follow-up sputum-positive' as their DST criterion and sputum smear microscopy status 'unknown' at the time of referral for DST were less likely to be initiated on treatment. The median delay to treatment initiation was 8 days (interquartile range 6-13). Patients referred for DST from medical colleges were more likely to face delays in treatment initiation. Conclusion: The Gujarat TB programme is performing well in initiating laboratory-confirmed MDR-TB patients on treatment. However, there is further scope for reducing delay.

Contexte : Dans le Gujarat, un état de l'ouest de l'Inde, même si la mise en route du traitement a été améliorée pour les patients ayant eu un diagnostic de tuberculose multirésistante (TB-MDR) dans le contexte des programmes, elle n'a pas encore atteint 100%.Objectif : Déterminer l'attrition avant traitement (c'est-à-dire un traitement pas mis en route dans les 6 mois suivant le diagnostic), le retard à la mise en route (>7 jours du diagnostic) et les facteurs associés parmi des patients TB-MDR diagnostiqués en 2014 dans cinq districts sélectionnés servis par deux structures de test génotypique de pharmacosensibilité (DST) et un centre de TB résistante au Gujarat.Schéma : Ceci a été une étude rétrospective de cohorte basée sur une revue de dossiers.Résultats : Sur 257 patients TB-MDR, l'attrition avant traitement a été constatée chez 20 patients (8% ; IC95% 5­12). Les patients ayant un « crachat de suivi positif ¼ comme critère de DST et un statut de microscopie de frottis de crachats « inconnu ¼ lors de la référence pour DST ont été moins susceptibles d'être mis sous traitement. Le délai médian de mise en route du traitement a été de 8 jours (intervalle interquartile 6­13). Les patients référés pour DST de centres hospitalières universitaires sont plus susceptibles de rencontrer des retards à la mise en route du traitement.Conclusion : Le programme TB du Gujarat est performant en mettant en route le traitement de TB-MDR confirmé par le laboratoire. Il reste cependant une marge d'amélioration en matière de réduction des délais.

Marco de referencia: Guyarat es un estado del occidente de la India donde se han logrado avances en la iniciación del tratamiento de los pacientes con diagnóstico de tuberculosis multirresistente (TB-MDR) en el marco programático, pero aún no se ha alcanzado el 100%.Objetivos: Determinar la tasa de abandono anterior al tratamiento (no haber iniciado tratamiento en un lapso de 6 meses después del diagnóstico), el retraso en la iniciación del tratamiento (>7 días después del diagnóstico) y los factores asociados en los pacientes diagnosticados con TB-MDR de cinco distritos escogidos de Guyarat atendidos por dos centros de pruebas genotípicas de sensibilidad a los medicamentos (DST) y un centro de tuberculosis resistente en el 2014.Método: Fue este un estudio de cohortes retrospectivo con examen de las historias clínicas.Resultados: De los 257 pacientes con diagnóstico de TB-MDR, se observó un abandono anterior al tratamiento en 20 casos (8%; IC95% 5­12). La probabilidad de iniciar el tratamiento era menor en los pacientes cuyo criterio para practicar las DST era 'seguimiento a la positividad del esputo' y su situación de la baciloscopia del esputo era desconocida en el momento de la remisión para las pruebas. La mediana del retraso en la iniciación del tratamiento fue 8 días (amplitud intercuartil 6­13). Los pacientes remitidos de las facultades de medicina para realizar las DST presentaban con mayor frecuencia retrasos en la iniciación del tratamiento.Conclusión: El desempeño del programa contra la TB de Guyarat es adecuado con respecto a la iniciación del tratamiento de los pacientes con TB-MDR confirmada por el laboratorio. Sin embargo, aún son necesarios progresos en esta esfera con el fin de acortar los retrasos.

Clinical implications of chromosomal abnormalities in 401 patients with myelodysplastic syndromes: a multicentric study in Japan.

It is well known that cytogenetic analysis in patients with myelodysplastic syndrome (MDS) provides information useful in determining their prognosis. Based on the chromosomal results obtained from 401 MDS patients by a multicentric study in Japan, we studied correlations between chromosomal findings and prognosis or leukemic transformation in MDS patients. Patients with complex aberrations (cytogenetic abnormalities at more than three chromosomes), of any subtype, had a poor prognosis; for example, > 60% of patients with refractory anemia (RA) showing complex aberrations died within one year, but only 11% of them developed leukemia. In patients with RA with ringed sideroblasts (RARS), > 70% of those with complex aberrations evolved into the leukemic phase and survived for less than one year, suggesting a biologic heterogeneity in RARS patients. By contrast, about 5% of patients with RA or RARS exhibiting chromosomal findings other than -7/7q-, +8, two aberrations, and complex aberrations, developed leukemia and had a favorable prognosis. Therefore, the presence of chromosome abnormalities alone in patients with RA or RARS is not a factor in predicting leukemic transformation or poor prognosis. In patients with refractory anemia with an excess of blasts (RAEB), the presence of chromosome aberrations at MDS diagnosis affected the occurrence of leukemic transformation (24% versus 43%), however, no particular difference was noted in patients with RAEB in transformation with regard to whether they had chromosome changes or not, and about 60% of them evolved into leukemia. The poor prognosis related to complex aberrations was consistently noted in all MDS subtypes or age-matched groups, indicating that this cytogenetic anomaly is an independent risk factor for a poor prognosis in MDS patients. The duration between MDS diagnosis and development of the leukemic phase and that between the latter and death were significantly shorter in patients with complex aberrations than those without this change. Although the clinical significance of certain chromosomal abnormalities differs among subtypes of MDS, a new scoring system for predicting prognosis by cytogenetic changes, in combination with hematologic parameters, was proposed.

Effects of an aplastic anemia urinary extract on mouse erythroid progenitor cells in vivo.

We investigated the in vivo effects of a crude extract from the urine of aplastic anemia patients (AA urinary extract) on erythroid precursor cells in the femoral bone marrow and spleens of normal adult mice. A single intraperitoneal injection of AA urinary extract induced a significant increase in the number of splenic erythroid burst-forming units (BFU-e) and erythroid colony-forming units (CFU-e) within 24 h after injection. We then injected pure recombinant erythropoietin (Epo) equivalent to the amount present in the urinary extract. This addition increased the number of splenic CFU-e by almost the same degree as the amount induced by the AA urinary extract 24 h after injection, but failed to elicit any change in the number of splenic BFU-e. In other studies, mice were injected with the same amount of lipopolysaccharide (LPS) and/or pure Epo as that present in the AA urinary extract. Experiments with Limulus amebocyte lysate-adsorbed (endotoxin-depleted) or nonadsorbed (endotoxin-containing) AA urinary extracts showed that endotoxin contamination interfered with the increase in numbers of marrow CFU-e and enhanced the increase in splenic CFU-e numbers induced by pure Epo or Epo activity in the AA urinary extract. The number of splenic BFU-e, however, was not affected by administration of LPS and/or Epo or by adsorbed endotoxin. These data suggest that AA urinary extract contains a stimulating activity for mouse splenic BFU-e, and that this activity is not attributable to the Epo activity or endotoxin contamination within the urinary extract.

Interleukin-6 and erythropoietin act as direct potentiators and inducers of in vitro cytoplasmic process formation on purified mouse megakaryocytes.

Mouse megakaryocytes were purified using a rabbit antimouse platelet serum, and magnetic beads were conjugated with an antirabbit IgG antibody. The purified cells were 95.8 +/- 1.2% megakaryocytes, and the recovery and viability of the megakaryocytes were 70 +/- 18.4%, and 80 +/- 13.4%, respectively. The effects of recombinant erythropoietin (Epo), interleukin-6 (IL-6), and IL-1 beta on these purified megakaryocytes were studied. Epo and IL-6 significantly increased DNA synthesis in these cells, but IL-1 beta did not. Similarly, both Epo and IL-6, but not IL-1 beta, increased the acetylcholinesterase (AchE) activity in the megakaryocytes. Epo and IL-6 stimulated the megakaryocytes to form cytoplasmic processes, which are considered to represent in vitro proplatelet formation. This process formation was inhibited by the addition of colchicine to the cultures. It was concluded that Epo and IL-6 are not only direct potentiators of megakaryocytes, but also inducers of in vitro cytoplasmic process formation on megakaryocytes.

In vitro and in vivo effects of MDP-Lys(L18) on mouse megakaryocyte progenitor cells (CFU-Meg).

We investigated the in vitro and in vivo effects of MDP-Lys(L18), a derivative of muramyl dipeptide (MDP), on megakaryocyte progenitor cells (megakaryocyte colony-forming units, CFU-Meg) in the mouse bone marrow and spleen. When CFU-Meg culture was performed with a suboptimum concentration (2%) of pokeweed mitogen-stimulated mouse spleen-conditioned medium (PWM-SCM), addition of 0.1-20 micrograms/ml of MDP-Lys(L18) increased the number of megakaryocyte colonies. The size of the megakaryocyte colonies (the number of megakaryocytes per colony) was also significantly increased by the addition of MDP-Lys(L18) under the same culture conditions in comparison with cultures without MDP-Lys(L18). MDP-Lys(L18) itself did not stimulate megakaryocyte colony formation without PWM-SCM, and it failed to enhance megakaryocyte colony formation in cultures with an optimum PWM-SCM concentration (10%). Furthermore, no effect of MDP-Lys(L18) was observed in cultures of phagocytic cell-depleted bone marrow cells. However, MDP-Lys(L18) enhanced megakaryocyte colony formation in cultures of T-lymphocyte-depleted bone marrow cells. The culture supernatant from a macrophage cell line, J774.1, plus MDP-Lys(L18) enhanced in vitro megakaryocyte colony formation in cultures with a suboptimum PWM-SCM concentration. Although interleukin 1 (IL-1)beta in the culture supernatant of J774.1 plus MDP-Lys(L18) was increased in a dose-dependent manner in response to MDP-Lys(L18), the effect of the culture supernatant was not blocked by an anti-IL-1 antibody, and IL-1 beta failed to enhance megakaryocyte colony formation in the presence of suboptimum PWM-SCM levels. The enhancement of megakaryocyte colony formation by MDP-Lys(L18) could be neutralized, however, by an anti-interleukin 6 (IL-6) antibody. Intraperitoneal administration of MDP-Lys(L18) (100 micrograms daily for 3 days) significantly increased the number of bone marrow and spleen megakaryocyte colonies at 24 to 72 h after the final injection. These in vitro and in vivo observations strongly suggest that MDP-Lys(L18) indirectly enhances the proliferation and differentiation of mouse CFU-Meg via colony-stimulating factor(s) other than IL-1, probably as a result of the stimulation of macrophages to produce IL-6.

Three stages of differentiation in mouse megakaryocyte progenitor cells (CFU-Meg).

Mouse megakaryocyte colonies developed in a fibrin clot culture system were morphologically classified into three types: immature homogeneous (IH), heterogeneous (H), and mature homogeneous (MH) colonies. The colony size (number of megakaryocytes per colony), the ploidy distribution of megakaryocytes in colonies, and the ratios of their progenitors (megakaryocyte colony-forming units, CFU-Meg) in the S-phase of the cell cycle were compared among the three types. Also, morphological changes in colonies were examined in time sequence. The mode of the number of megakaryocytes per colony on day 6 of culture was greater than 64, between 17 and 32, and between 4 and 8, in IH, H, and MH colonies, respectively. The mean DNA content of megakaryocytes was 3.1N, 4.2N, and 7.2N on day 6 of incubation, in IH, H, and MH colonies, respectively, these values being significantly different from each other (p less than 0.001). The mean proportion of CFU-Meg synthesizing DNA was 21.9%, 26.0%, and 48.6% for CFU-Meg forming IH, H, and MH colonies, respectively (p less than 0.01; IH- versus MH-CFU-Meg and H- versus MH-CFU-Meg). Successive observation of the colony morphology from days 5 to 11 of culture suggested a transformation of the colony types from IH colony to MH colony through H colony. These observations indicate that the three types of megakaryocyte colonies classified on the basis of their morphological features reflect various stages of differentiation of mouse CFU-Meg.

Cloning of murine megakaryocyte progenitor cells in a fibrin clot culture system.

A fibrin clot culture system was applied to the cloning of mouse megakaryocyte colony-forming cells (CFU-Meg). The culture medium in this new method consists of Iscove's minimal essential medium containing fetal bovine serum, bovine fibrinogen, bovine thrombin, and pokeweed mitogen-stimulated mouse spleen cell-conditioned medium (PWM-SCM). CFU-Meg colony frequency with 10% PWM-SCM was maximal on days 5-6 of culture. Plating efficiencies averaged 36.1 +/- 3.9 and 51.9 +/- 6.0 per 1.5 X 10(5) BDF1 bone marrow cells and 1.0 X 10(6) spleen cells, respectively. The addition of bovine serum albumin to the culture medium had no effect on the efficiency of megakaryocyte colony growth in this culture system. This simplified and reproducible culture system supported not only the growth of colonies composed of megakaryocytes in "synchronous maturation," but also so-called "heterogenous" megakaryocyte colonies composed of cells in all stages of maturation.

In vitro and in vivo effects of KT6352, a derivative of indolocarbazole compounds, on murine megakaryocytopoiesis.

We investigated the in vitro and in vivo effects of KT6352, a derivative of indolocarbazole compound, on murine megakaryocytopoiesis. When serum-free megakaryocyte (Meg) colony assay was performed with 100 U/mL of recombinant mouse interleukin-3 (rmIL-3), the addition of 1x10(-11)M to 1x10(-9)M of KT6352 increased the number of Meg colonies. An additional increase of Meg colonies by KT6352 was observed in the serum-free culture containing rmIL-3 plus recombinant mouse interleukin-6 or rmIL-3 plus recombinant mouse stem cell factor. KT6352 did not stimulate Meg colony formation without rmIL-3. When KT6352 was administered intraperitoneally to normal BALB/c male mice at a dose of 10 mg/kg daily for 5 consecutive days, a 2.1-fold increase in the platelet count was observed on day 14, and the prolonged thrombocytopoiesis was detectable from 9 to 27 days after KT6352 administration. A marked increase in the white blood cell count was also observed from 5 to 14 days after KT6352 treatment. Before the gradual increase of platelet counts, 8 days after KT6352 administration, a marked increase in the number of colony-forming units of megakaryocytes (CFU-Megs) in bone marrow and spleen was observed, and a substantial increase in the number of splenic CFU-Megs was observed 14 and 23 days after KT6352 administration. Bone marrow Meg ploidy analysis by two-color flow cytometry showed a shift in the modal ploidy class from 16 to 32 and an increase in the frequency of 64 cells in KT6352-treated mice. These results suggest a possible therapeutic benefit of KT6352 in the management of thrombocytopenia.

Effects of c-mpl ligand on cytoplasmic maturation of murine megakaryocytes and on platelet production.

To test the hypothesis that the c-mpl ligand is not a primary factor in thrombocytopoiesis, we investigated the biological effects of recombinant human (rh) c-mpl ligand on differentiation of murine progenitor cells and on maturation of the cultured murine megakaryocytes under serum-free conditions on the basis of ploidy distribution, megakaryocyte/platelet-specific surface antigen CD 61 [glycoprotein (GP) IIIa], and cytoplasmic acetylcholinesterase (AchE) expression in vitro. In addition, we studied the effect of c-mpl ligand on proplatelet formation (PPF) by murine mature megakaryocytes. AchE was less strongly expressed in cultured megakaryocytic cells stimulated by c-mpl ligand than in those stimulated by recombinant murine (rm) IL-3 + rh IL-6 during the differentiation of progenitor cells. Less CD 61 was expressed by c-mpl ligand during both the differentiation of progenitor cells and the maturation of megakaryocytes compared with that by rm IL-3 + rh IL-6. Endomitosis, however, was more stimulated by c-mpl ligand than by rm IL-3 + rh IL-6 under both conditions. Furthermore, PPF of mature megakaryocytes was not stimulated by c-mpl ligand. These results indicate that c-mpl ligand stimulates the nuclear development of megakaryocytic cells but that it does not stimulate cytoplasmic maturation and PPF as much as IL-6. These data strongly suggest that c-mpl ligand is not a primary factor in platelet pro-duction. (J Histochem Cytochem 46:49-57, 1998)

Purification of proplatelet formation (PPF) stimulating factor: thrombin/antithrombin III complex stimulates PPF of megakaryocytes in vitro and platelet production in vivo.

In this study, the protein which stimulates proplatelet formation (PPF) of megakaryocytes was purified from normal human plasma using 7 steps procedures. Two different protease inhibitors were identified based on their amino acid sequences, i.e. antithrombin III (AT III) and C1 inhibitor. They were included in high density lipoprotein (HDL). HDL was necessary for AT III to be active in PPF in vitro. The biological effects of the AT III/HDL or thrombin-AT III (TAT)/HDL were studied in vitro. PPF of murine megakaryocytes was stimulated by negative control (BSA) (1.8 +/- 0.3%), AT III (2.0 +/- 0.4%), HDL (1.2 +/- 0.9%), AT III/HDL (14.8 +/- 2.1%) or TAT/HDL (23.3 +/- 3.5%), respectively. TAT/HDL also had a synergistic effect with the mpl ligand, judging by the acetylcholinesterase (AchE) expression of murine megakaryocytes (2.7 fold increase). In vivo subcutaneous administration of AT III alone or TAT for 3 days significantly stimulated thrombocytosis (136% and 144%, respectively, p<0.05) and AT III/HDL showed rapid and further stimulation (150%, p <0.01). These results and the previous studies indicate that megakaryocytopoiesis is regulated by the mpl ligand, while a protease/protease inhibitor complex such as TAT, which is involved in the coagulation cascade associated with platelet consumption, might be one of the regulators in platelet production.

Flow cytometric analysis of aberrant antigen expression of blasts using CD45 blast gating for minimal residual disease in acute leukemia and high-risk myelodysplastic syndrome.

To evaluate the availability of quantification of blasts with aberrant antigen expression (AAE) using CD45 gating for minimal residual disease (MRD), 15 patients with acute leukemia (AL) and myelodysplastic syndrome (MDS) were studied. In patients with complete remission (CR), the frequency of blasts with AAE (%AAE) by CD45/side scatter (SSC) gating was significantly higher than that by the traditional forward scatter (FSC)/SSC combination (median, 4.1 vs. 0.3%, P<0.0001). We also demonstrated two representative cases, in which leukemia relapse could be predicted before 3 weeks and the early treatment for MRD could be performed for MRD after allogeneic bone marrow transplantation (BMT). These results indicate that this procedure is very useful for the evaluation of the quality of CR.

The relationship between the site of breakpoints within the bcr gene and thrombopoiesis of Philadelphia-positive chronic myelocytic leukemia.

We determined the position of the breakpoint within the bcr gene in 22 patients with Philadelphia-positive chronic myelocytic leukemia using conventional Southern-blots and analyzed its relationship to thrombopoiesis. After DNA digestion with restriction endonucleases (Hind III, Bam HI and Bgl II), we localized the breakpoint in bcr using two genomic probes. The location of the breakpoint within the bcr was assigned to one of five zones. Breakpoints in zones 1 and 2 were grouped as "5"', and those in zones 3, 4 and 5 as "3'". Thus we subdivided patients with bcr rearrangements into those with genomic breaks at either 5' or 3' of the Bam HI site, just upstream of exon 3. Nine patients had 5' breakpoints and 13 patients had 3' breakpoints. The platelet counts of 3' patients were significantly higher than those of 5' patients (1395 vs 274 x 10(9)/l; p less than 0.03). The megakaryocyte counts from bone marrow histological sections in 3' patients (n = 12) and 5' patients (n = 7) were 63.4/mm2 and 19.5/mm2, respectively, with a significant difference of p less than 0.006. The mean number of megakaryocyte progenitor cells assayed by in vitro cloning was 128.3/2 x 10(5) bone marrow cells for 3' patients (n = 7) compared with 46.3 for 5' patients (n = 4). These results suggest that Philadelphia-positive CML patients with 3' breakpoints have higher thrombopoietic activity than patients with 5' breakpoints.

Successful treatment of two patients with primary cardiac malignant lymphoma.

We describe two patients with primary cardiac malignant lymphoma involving the right atrium and superior vena cava, resulting in intractable right cardiac failure and superior vena cava syndrome. Patients were diagnosed by surgical myocardial biopsy and were treated with combination chemotherapy for non-Hodgkin's lymphoma. Each attained a marked response, and hence avoided sudden death from tricuspid atresia. Both have remained alive for more than 21 and 34 months, respectively, and continue intermittent combination chemotherapy.

Clinical and cytogenetic findings of myelodysplastic syndromes showing hypocellular bone marrow or minimal dysplasia, in comparison with typical myelodysplastic syndromes.

Hematologic and cytogenetic data were collected on 401 myelodysplastic syndromes (MDS) patients in Japan and their clinical relevance was analyzed. More than 50% of the MDS patients with hypocellular bone marrow had > 5% marrow blasts at the time of MDS diagnosis and frequently had complex aberrations (chromosome changes at three or more regions). They showed peripheral blood findings resembling those of aplastic anemia, but progression into leukemic phase and the prognosis tended to mimic typical MDS. In MDS patients with minimal dysplasia, hematologic parameters were different from those of aplastic anemia. However, the low incidence of leukemic transformation and the favorable prognosis was similar to that of aplastic anemia. More than 90% of the patients in this group had refractory anemia and had normal karyotypes. Thus differential diagnosis from a low grade aplastic anemia is important, since this type of MDS might have a clonal nature as well. In conclusion, although some hematologic and clinical deviations are noticed in MDS with hypocellular marrow or minimal dysplasia, these MDS subtypes might constitute marginal forms of MDS.

Efficacy of a new formulation of lenograstim (recombinant glycosylated human granulocyte colony-stimulating factor) containing gelatin for the treatment of neutropenia after consolidation chemotherapy in patients with acute myeloid leukemia.

The efficacy and safety of a new formulation of lenograstim (recombinant glycosylated granulocyte colony-stimulating factor) prepared by switching the stabilizer from human serum albumin (HSA) to gelatin was investigated for the treatment of neutropenia after consolidation chemotherapy in patients with acute myeloid leukemia (AML). The results obtained in the study using the gelatin-containing formulation (gelatin-lenograstim) were retrospectively compared to those obtained from a placebo-controlled double-blind randomized study (AML-DBT) using the HSA-containing formulation (HSA-lenograstim). The median time of neutrophil recovery to > or = 1000/mm3 was significantly shorter in the gelatin-lenograstim group (14 days) than in the placebo group (21 days, P = .0001), and there was no significant difference between the gelatin-lenograstim group and the HSA-lenograstim group (14.5 days of AML-DBT, P = .5462). The incidences of febrile neutropenia were significantly reduced in the gelatin-lenograstim group (24/43, 55.8%) compared to the placebo group (58/64, 90.6%, P < .0001). The incidence of fever and antibiotic use was also significantly lower in the gelatin-lenograstim group (69.8% and 83.7%, respectively) than in the placebo group (92.2%, P = .0034, and 96.9%, P = .0285, respectively). However, between the 2 groups there were no differences in the number of patients who had infectious episodes. No serious adverse drug reactions ascribed to gelatin-lenograstim were encountered. These results demonstrate that gelatin-lenograstim exerted beneficial effects in the acceleration of neutrophil recovery and in the reduction of fever, febrile neutropenia, and antibiotic use, and its efficacy was equivalent to HSA-lenograstim. Therefore, we concluded that the gelatin-lenograstim formulation, which offers no risk of virus contamination and can be stored at room temperature, is more beneficial than the HSA-lenograstim formulation.

TdT-Positive, SmIg-Negative B Precursor Cell Leukemia with Burkitt Morphology: A Case Report.

The phenotypic marker profile and molecular analysis of ALL with typical FAB-L3 morphology of a 66-year-old lady are described. We detected the phenotypic expression of B cell-associated antigens (B4, BA1, HLA-DR) and terminal deoxynucleotidyl transferase, but not of surface and cytoplasmic immunoglobulin or cALLA antigen. Molecular analysis revealed that immunoglobulin heavy chain gene was in the rearranged form and that T cell ß-receptor gene was in the germ-line form Cytomorphology, immunophenotype and genotype indicated that her leukemic cells had characteristics of B-cell precursor ALL of Burkitt type. Cytogenetic investigation showed a normal karyotype, and neither rearrangement nor amplification of c-myc gene was detected To the best of our knowledge this type of TdT-positive, Smlg-negative FAB-L3 with immunological and molecular confirmation has as yet not been reported.

[Autoimmune hemolytic anemia associated with adult-onset Still's disease].

A 37-year-old woman was admitted to our hospital in March 1995 because of high fever and cervical lymph node swelling. She had received prednisolone (PSL) therapy for autoimmune hemolytic anemia (AIHA). Laboratory examinations revealed increased serum levels of liver enzymes and C-reactive protein. Levels of rheumatoid factor and anti-nuclear antibody were within normal limits. On the basis of these criteria, she was diagnosed as having adult-onset Still's disease (AOSD). Although her fever ameliorated and physical symptoms disappeared immediately after a moderate dose of PSL, hemolysis recurred when attempts were made to reduce or withdraw the steroid. The patient is now receiving low-dose PSL therapy and is free of AIHA and AOSD.

[Effects of recombinant human alpha-2b and gamma interferons on bone marrow megakaryocyte progenitors (CFU-Meg) from patients with chronic myelocytic leukemia].

The effects of recombinant human interferon (IFN) alpha-2b and gamma on the bone marrow megakaryocyte progenitors (CFU-Meg) were compared between eight patients in the chronic phase of Ph1-positive chronic myelocytic leukemia (CML) and five hematologically normal patients. CFU-Meg was assayed in plasma clot culture added with phytohemagglutinin-stimulated leukocyte-conditioned medium as a source of colony stimulating activity. The average count of CFU-Meg colonies formed from the bone marrow of CML patients was 5.5 times that of normal controls. Spontaneous CFU-Meg colonies were grown in seven of eight CML patients, but in none of five controls. Colony formation by CFU-Meg in CML as well as normal bone marrow was suppressed by the two preparations of IFN in a dose dependent fashion. Their suppressive influence on colonies from CFU-Meg was comparable between CML and normal bone marrow at lower concentrations, but was less marked for CML than normal bone marrow at higher concentrations. The formation of CFU-Meg colonies from CML bone marrow was more severely suppressed by IFN-gamma than IFN-alpha-2b. Depletion of either T lymphocytes or adherent cells from the CML bone marrow cells diminished the suppressive effects of IFN-gamma, but had no influence on the effects of IFN-alpha-2b.