bluuebunny/crossref_metadata_2025 huggingface dataset:
The effects of environmental variation on bryophytes at a regional scale
Authors: Vanderpoorten Alain , Engels Patrick
Published: March 2003

The distribution of bryophytes in central Belgium was investigated using species grid‐mapping superimposed on a series of maps which included information on soil conditions and land use. Our objectives were to assess the influence of environmental variation on the bryoflora at a regional scale, to examine how bryophytes respond to environmental variation, and to assess the extent to which species ecological and life‐history traits determine the accuracy of the predictability of species occurrence in order to provide comprehensive lists of species based on environmental conditions. The first two axes of a correspondence analysis (CA) of the floristic data explained 14.6% of the total χ2. CA1 was significantly correlated with loamy‐sandy soils on a sand layer (r=−0.74, p<0.001), forest cover (r=−0.80, p<0.001), loamy soils (r=0.79, p<0.001), and agricultural fields cover (r=0.61, p<0.001). CA2 had a low but significant correlation coefficient with pebbly soils cover (r=0.38, p<0.001). The probability of occurrence of 59% of the investigated species could be significantly predicted by logistic regression from the sets of environmental variables. About 55% of the species exhibited an increasing probability of occurrence with increasing forest cover and loamy‐sandy soils cover, 1% with agricultural fields and loamy soils cover, and 3% with pebbly soils cover. The predictability of species occurrence varied as a function of four life‐history traits (minimum spore size, life expectancy, type of gametophyte and papillose leaf cell walls) and three ecological traits (indicator values of light, temperature and soil acidity). The most predictable species, including a number of leafy liverworts, were characteristic for acidic, fresh and shaded conditions and displayed a strong preference for forest habitats. Taxa with limited predictability included epiphytes and mosses characteristic of pebbly soils due to the ability of these species to efficiently disperse and adapt to various ecological conditions. Species for which the distribution range could not successfully be predicted were either ubiquitous, characteristic for ephemeral habitats, or highly successful in a very common habitat.
Geographic range, turnover rate and the scaling of species diversity
Authors: Arita Héctor T. , Rodríguez Pilar
Published: March 2003

The study of the relative roles of local and regional processes in determining the scaling of species diversity is a very active field in current ecology. The importance of species turnover and the species‐range‐size frequency distributions in determining how local and regional species diversity are linked has been recognised by recent approaches. Here we present a model, based on a system of fully nested sampling quadrats, to analyse species diversity at several scales. Using a recursive procedure that incorporates increasingly smaller scales and a multiplicative formula for relating local and regional diversity, the model allows the simultaneous depiction of alpha, beta and gamma diversity in a single “species‐scale plot”. Species diversity is defined as the number of ranges that are intersected by sampling quadrats of various sizes. The size, shape and location of individual species ranges determine diversity at any scale, but the average point diversity, measured at hypothetical zero‐area localities, is determined solely by the size of individual ranges, regardless of their shape and location. The model predicts that if the species‐area relationship is a power function, then beta diversity must be scale invariant if measured at constant scale increments. Applying the model to the mammal fauna of four Mexican regions with contrasting environmental conditions, we found that: 1) the species‐range‐size frequency distribution at the scale of the Mexican regions differs from the log‐normal pattern reported for the national and continental scales. 2) Beta diversity is not scale‐invariant within each region, implying that the species‐area relationship (SAR) does not follow a power function. 3) There is geographic variation in beta diversity. 4) The scaling of diversity is directly linked to patterns of species turnover rate, and ultimately determined by patterns in the geographic distribution of species. The model shows that regional species diversity and the average distribution range of species are the two basic data necessary to predict patterns in the scaling of species diversity.
Dynamics or constancy inSphagnumdominated mire ecosystems? A 40‐year study
Authors: Gunnarsson Urban , Malmer Nils , Rydin Håkan
Published: March 2003

Traditionally mire ecosystems (especially bogs) have been viewed as stable systems with slow changes in the vegetation over time. In this study the mire Åkhultmyren, south‐central Sweden was re‐investigated in 1997 after 40 yr of continued natural development. The results show a high degree of dynamics in aSphagnumdominated bog and fen. Altogether 97 vascular plant and bryophyte species were recorded in the two inventories of the bog and poor fen vegetation. pH and electrical conductivity in the mire water were also surveyed. In 1997 we found 10 new species and that 8 species had disappeared since 1954 but the over‐all mean number of species per plot (size 400 m2) had hardly changed. However, 21% of the species increased and 21% decreased significantly in frequency. Most of the species that decreased in frequency were low‐grown vascular plants, most common in wet microhabitats. Vascular plant species that increased in frequency included trees (defined as >1.3 m in height) and were generally taller than the unchanged or decreasing species. The frequency of dwarf shrubs and hummock bryophytes increased too. Areas with an initial pH of 4.5–5.0 showed the strongest decrease in pH, coinciding with an enlarged distribution of someSphagnumspecies. The species diversity increased on the bog, but decreased in the wettest parts of the fen, where the pH also decreased. Species with unchanged or increasing frequency often showed high capacity to colonise new plots. On average the sum of gains and losses of species in the plots in 1997 was ca 50% of the species number in 1954. The vegetation changes indicate a drier mire surface and an increased availability of nitrogen. The increased tree cover may have triggered further changes in the plant cover.
Elevated serum levels of soluble CD44 variant 6 are correlated with shorter survival in aggressive non‐Hodgkin's lymphoma
Authors: Sasaki Kyoko , Niitsu Nozomi
Published: March 2003

Abstract: A variant form of CD44 that has additional amino acids in the common protein backbone (CD44‐v6) seems to play a role in the metastasis of malignancies. We measured soluble CD44‐v6 (sCD44‐v6) by ELISA in 201 patients with malignant lymphoma. The sCD44‐v6 level was significantly elevated in patients with non‐Hodgkin's lymphoma (NHL) (n=184). The sCD44‐v6 level was correlated significantly with the standard sCD44 and soluble interleukin‐2 receptor levels, but only weakly with serum lactate dehydrogenase (LDH). In 149 patients with aggressive NHL, the sCD44‐v6 level was elevated in the subgroups with a high LDH level, stage III/IV disease, T‐cell lymphoma, and high–intermediate or high risk group as identified by the International Prognostic Index (IPI). When the sCD44‐v6 level was 800 ng/ml the overall survival rate was significantly decreased (p=0.0001). In the low+low–intermediate risk group (IPI) both overall survival rates (log‐rank p=0.0005, Wilcoxon p=0.002) were significantly decreased when the sCD44‐v6 level was 800 ng/ml. In multivariate analysis, sCD44‐v6 was shown to be independent of the five prognostic factors in the IPI (age, performance status, number of extranodal sites, Ann Arbor stage and LDH level), so it may be useful for predicting the outcome of aggressive NHL.
Normal ranges for packed cell volume and hemoglobin concentration in adults: relevance to ‘apparent polycythemia’
Authors: Fairbanks Virgil F. , Tefferi Ayalew
Published: March 2003

Abstract: Published data from Europe and North America indicate that for non‐iron‐deficient adult Caucasian males, the normal mean packed cell volume (PCV) is 0.46 and the 2.5–97.5 percentile interval is04.0–0.53. Corresponding values for adult Caucasian females are: mean PCV 0.42; 2.5–97.5 percentile interval 0.36–0.48. It is not usually appropriate to undertake studies of polycythemia in adult Caucasian males with PCV<0.55 (Hb Conc.<180 g/L) or in adult Caucasian females with PCV<0.50 (Hb Conc. <16.5 g/L). Application of this principle will reduce the number of inappropriate and costly studies that would otherwise be performed in patients whose PCV values are only in the upper percentiles of the normal range, and will help to avoid misdiagnoses and therapeutic misadventures.
Lenograstim for the treatment of neutropenia in patients receiving ganciclovir for cytomegalovirus infection: a randomised, placebo‐controlled trial in AIDS patients
Authors: Dubreuil‐Lemaire M‐L. , Gori A. , Vittecoq D. , Panelatti G. , Tharaux F. , Palisses R. , Gharakhanian S. , Rozenbaum W. ,   GCS 309 European Study Group
Published: March 2003

Abstract: This phase IIa, randomised, single‐blind, placebo‐controlled study was conducted to determine the dose of recombinant human granulocyte colony‐stimulating factor (lenograstim) suitable for use in AIDS patients. The study was conducted at 27 European AIDS/HIV centres, and recruited 69 AIDS patients with an initial episode or relapse of cytomegalovirus infection (neurological site excluded) and an absolute neutrophil count (ANC) 1.0×109/L upon diagnosis or between days 1 and 12 of ganciclovir (GCV) treatment. The patients were randomised to placebo (n=14) or one of four lenograstim arms: 150 µg/m2/d (the standard onco‐haematology dose, n=13) or 100 (n=13), 50 (n=15), or 25 µg/m2/d (n=14). In all groups, the planned dose of GCV was 10mg/kg/d for 21 d. Median ANC at weeks 2 and 3 was significantly higher in each lenograstim group than in the placebo group (p=0.05). At week 3, median ANC (×109/L) was 0.7 in the placebo group, compared with 6.0, 7.4, 4.5, and 2.0 in the 150, 100, 50, and 25 µg2/d lenograstim groups, respectively. Median ANC was not significantly different between the 150, 100, and 50 µg/m2/d lenograstim groups at any time point, but significantly higher in the 50 than in the 25 µg/m2/d group at weeks 2 (p=0.05) and 3 (p=0.02). Lenograstim was generally well tolerated, leading to no severe adverse events. In conclusion, lenograstim 50µg/m2/d is suitable for the treatment of ganciclovir‐induced neutropenia and is safe. These results should help the physician choose an optimal and cost‐efficient regimen for patients with AIDS‐related neutropenia when rHuG‐CSF support is indicated.
Thrombopoietin activates the growth of megakaryoblasts in patients withchronic myeloproliferative disorders and myelodysplastic syndrome
Authors: Hashimoto Shigeo , Toba Ken , Fuse Ichiro , Watanabe Kenichi , Takahashi Hidenobu , Abe Takashi , Yano Toshio , Koike Tadashi , Takahashi Masuhiro , Aizawa Yosifusa
Published: March 2003

Abstract: The effects of thrombopoietin (TPO) on cell proliferation and differentiation, and the relation between these effects and the expression of c‐mpl on leukemia cells were studied in seven acute myelogeneous leukemia cell lines and seven myelogeneous blast cell preparations from patients with chronic myeloproliferative disorders (CMPDs) and myelodysplastic syndrome (MDS). Among the leukemia cells, five preparations of megakaryoblastic leukemia cells from patients and one megakaryoblastic cell line, CMK 11.5, proliferated in response to TPO in vitro. CMK 11.5 and the blastic cells from one patient diagnosed with MDS with myelofibrosis differentiated with increasing expression of CD41a in response to TPO. However, TPO had no effect on the cells lacking megakaryocytic characteristics. Some patients with CMPD and MDS develop acute transformation with blasts demonstrating megakaryocytic features, and some of these cells show growth in response to TPO. Therefore, in vivo administration of TPO should be considered carefully for patients with CMPD or MDS, since TPO may induce leukemic cell proliferation.
Two‐phase liquid culture system models normal human adult erythropoiesis at the molecular level
Authors: Pope Sharon H. , Fibach Eitan , Sun Jun , Chin Kyung , Rodgers Griffin P.
Published: March 2003

Abstract: We have studied the patterns of expression of various genes during maturation of normal human adult erythroid precursors cultured in a two‐phase liquid culture method. In the first phase, peripheral blood mononuclear cells are cultured for one week in the presence of a combination of growth factors, but not erythropoietin (Epo). In Phase II, Epo is included in the medium. Cell samples were taken throughout phase II, and expression of globins, transcription factors, and cytokine receptors was assayed by RT‐PCR and quantified by phosphor imaging. We have divided phase II into stages: early (days 0–5), intermediate (days 6–10) and late (days 11–15) and measured maximum expression of each gene. During early phase II, γ‐globin, Sp1, and GATA‐2 mRNAs were expressed at their highest levels. As the cells matured during the intermediate period, GATA‐2 levels remained high, and then declined, while the transcription factors GATA‐1, EKLF, NF‐E2, and the Epo receptor (EpoR) reached maximum expression. In late phase II, β‐globin increased and reached its maximum level of expression. This erythroid culture system appears to recapitulate normal adult erythropoiesis at the molecular level, and thus may be a suitable model to examine the molecular basis of severe congenital or acquired disorders oferythropoiesis.
Biochemical markers of bone metabolism reflect osteoclastic and osteoblastic activity in multiple myeloma
Authors: Abildgaard N. , Glerup H. , Rungby J. , Bendix‐Hansen K. , Kassem M. , Brixen K. , Heickendorff L. , Nielsen J. L. , Eriksen E. F.
Published: March 2003

Abstract: In order to evaluate the use of recently developed assays of bone metabolism in multiple myeloma we performed a histomorphometric study of bone biopsies in 16 myeloma patients. Furthermore, we measured the levels of interleukin‐6 (IL‐6), soluble IL‐6 receptor (IL‐6sR), IL‐1β, tumour necrosis factor (TNF) α, TNFβ, and transforming growth factor (TGF) β in marrow plasma aspirated from the biopsy area. Markers of bone resorption: The N‐terminal telopeptide of collagen I (Ntx) in urine showed a strong positive correlation with the dynamic histomorphometric indices of bone resorption (r=0.68–0.72). Slightly weaker correlations were observed between the dynamic indices of bone resorption and the C‐terminal telopeptide of collagen I (ICTP) in serum (r=0.57–0.62) and deoxypyridinoline (Dpyr) in urine (r=0.54), whereas urinary pyridinoline (Pyr) did not correlate with the histomorphometric findings. Markers of bone formation: Serum C‐terminal propeptide of procollagen I (PICP) and serum bone‐specific alkaline phosphatase (bAP) showed significant correlations with the dynamic parameters of bone formation (r=0.57–0.58), whereas serum osteocalcin and serum total AP did not. Cytokines: Highly significant correlations were observed between marrow IL‐6 and rates of bone resorption and activation frequency (r=0.76–0.82) and with serum ICTP (r=0.63). Minor, but also significant correlations were observed between the resorptive indices and IL‐6sR and IL‐1β. The data indicate that measurements of the biochemical markers of bone metabolism may be useful in monitoring myeloma bone disease, and might thus be of use for dose titration of bisphosphonate therapy.
Human immunodeficiency virus‐associated malignant lymphoma in eastern Denmark diagnosed from 1990 to 1996: clinical features, histopathology, and association with Epstein‐Barr virus and human herpesvirus‐8
Authors: Boye Hansen Per , Penkowa Milena , Kirk Ole , Skinhøj Peter , Pedersen Court , Lisse Ida , Kiss Katalina , Zhou Xiao‐ge , Hamilton‐Dutoit Stephen J.
Published: March 2003

Abstract: The clinicopathological features of human immunodeficiency virus (HIV)‐associated lymphoma were investigated in a retrospective study of 85 adult patients in eastern Denmark diagnosed during the period 1990–1996. The possible pathogenetic role of Epstein‐Barr virus (EBV) and human herpesvirus 8 (HHV‐8) in these tumours was also studied. Seventy patients (82%) presented with extranodal disease and 26 (31%) had CNS involvement at diagnosis. Diffuse large cell B‐cell lymphoma was the most frequent histological subtype, comprising 65 of 79 cases available for microscopic re‐evaluation (82%) and including 20 of 23 evaluable patients with CNS lymphoma (87%). EBV RNA was demonstrated by in situ hybridization in 51 of 65 evaluable tumours (79%) and in 14 of 16 cases (88%) with CNS‐lymphoma. Three cases showed a T‐cell phenotype. The presence of HHV‐8 DNA was analysed by PCR in 32 cases. A strong band consistent with tumour cell infection was detected in only one case, weaker bands being seen in 4 cases. None of these patients had primary effusion lymphomas. In conclusion, Danish AIDS‐related lymphomas are of predominantly high‐grade B‐cell type with extranodal localization and atypical presentation. Our results provide further evidence that EBV plays a major role in the pathogenesis of large cell AIDS‐related lymphoma, whereas HHV‐8 does not appear to contribute significantly to the development of solid lymphomas in this group of patients.
Cytogenetic and FISH studies of a single center consecutive series of 152 childhood acute lymphoblastic leukemias
Authors: Andreasson Patrik , Höglund Mattias , Békássy Albert N. , Garwicz Stanislaw , Heldrup Jesper , Mitelman Felix , Johansson Bertil
Published: March 2003

Abstract: Between 1977 and 1996, cytogenetic investigations were performed on 182 childhood (16 yr) acute lymphoblastic leukemias (ALL), constituting 94% (182 of 194) of all ALL patients diagnosed and treated at the Departments of Pediatrics, Lund and Malmö University Hospitals, Sweden, during these two decades. The cytogenetic analyses were successful in 152 cases (84%). The failure rate was higher for the ALL investigated before 1987 (30% vs. 4%, p<0.0001), and also the incidence of cytogenetically normal cases was higher during 1977–86 (43% vs. 25%, p<0.05). Clonal chromosomal abnormalities were found in 103 (68%) ALL. Structural rearrangements were detected, by chromosome banding alone, in 76 cases (50%). Fluorescence in situ hybridization (FISH) was used to identify cases with t(12;21), 11q23 rearrangements, and 9p deletions, using probes for ETV6/CBFA2, MLL, and CDKN2A/B, in 72 cases from which cells in fixative and/or unstained metaphase preparations were available. In total, the most common structural rearrangements were del(9p) (17%), t(12;21) (15%), del(6q) (8%), and MLL rearrangements (4%). Six (32%) of nineteen cytogenetically normal ALL analyzed by FISH harbored cryptic abnormalities; three displayed t(12;21) and four had del(9p), one of which also carried a t(12;21). Five (45%) of the t(12;21)‐positive ALL showed +der(21)t(12;21) or ider(21)(q10)t(12;21), resulting in the formation of double fusion genes. Among the more rare aberrations, eight structural rearrangements were identified as novel recurrent ALL‐associated abnormalities, and nine cases harbored rearrangements previously not reported. Sixteen cases displayed karyotypically unrelated clones at different investigations. Seven ALL (5%) showed simple chromosomal changes, unrelated to the aberrations detected at diagnosis, during morphologic and clinical remission, and in all but one instance the patients remained in remission, with the abnormal clone disappearing in subsequent investigations. This indicates that the emergence of novel clonal chromosomal aberrations during remission in childhood ALL is rather common and does not by necessity predict a forthcoming relapse.
Plasma‐soluble Fas (APO‐1, CD95) and soluble Fas ligand in immune thrombocytopenic purpura
Authors: Yoshimura Chie , Nomura Shosaku , Nagahama Minori , Ozaki Yoshio , Kagawa Hideo , Fukuhara Shirou
Published: March 2003

Abstract: We investigated the levels of various cytokines and soluble factors in ITP patients, in order to determine the influence of these factors on the pathogenesis of ITP. We found increases in IL‐2, IL‐6, IFN‐γ, and M‐CSF levels in ITP patients compared with those in healthy individuals. On lymphocyte phenotype analysis, we found no clear difference in total T cell population (CD2+ CD19− cells) or cytotoxic T cell frequency (CD8+ CD11b− cells) between these two groups. The frequency of helper/inducer T cells (CD4+ CD8− cells) was decreased in ITP patients. There was a significant increase in activated T cells (CD3+ HLA‐DR+ cells) in ITP patients. Furthermore, frequencies of NK cells of potent activity (CD16+ CD56+ cells) were significantly elevated in ITP patients. Seventeen of the 54 ITP patients (31.5%) had elevated levels of sFas, and 11 of the 54 patients (20.4%) of sFasL. In addition, a significant increase of sFasL was observed in sFas‐positive ITP patients, and in these patients the sFasL level was correlated with that of sFas (r=0.687, p<0.01). We found significant increases in IL‐2 and sIL‐2R levels in sFas‐positive ITP patients. For other factors examined, however, there were no differences in level between sFas‐positive and‐negative ITP patients. Percentages of activated T cells (CD3+ and HLA‐DR+ cells) and NK cells (CD16+ and CD56+ cells) were significantly higher in sFas‐positive ITP patients than in sFas‐negative ITP patients. These findings suggests that the pathogenesis of ITP includes alteration of the Fas/FasL pathway.
Cladribine in combination with mitoxantrone and cyclophosphamide(CMC) in the treatment of heavily pre‐treated patients with advanced indolent lymphoid malignancies
Authors: Robak Tadeusz , Góra‐Tybor Joanna , Lech‐Marańda Ewa , Błoński Jerzy Z. , Kasznicki Marek
Published: March 2003

Abstract: The aim of our study was to determine the effectiveness and toxicity of combined chemotherapy consisting of cladribine (2‐chloro‐deoxyadenosine, 2‐CdA), mitoxantrone and cyclophosphamide (CMC regimen) in the treatment of refractory or relapsed indolent lymphoproliferative disorders. The treatment course consisted of 2‐CdA given at a dose of 0.12 mg/kg/24 h in a 2‐h intravenous infusion for 5 (CMC5) or 3 (CMC3) consecutive days, mitoxantrone 10 mg/m2 on day 1 and cyclophosphamide 650 mg/m2/iv on day 1. Thirty‐three patients (19 with B‐CLL and 14 with LG‐NHL) entered the study and all of them were eligible. Twenty patients (60.6%) were recurrent after prior therapy and 13 (39.4%) had refractory disease. All patients received 5 or more cycles of chemotherapy before CMC treatment. Twenty‐one patients were treated with CMC5 regimen and 12 with CMC3 regimen. The overall response rate, including CR and PR, was 48.6% (95% CI 32–66). There were no differences in the frequency of responses between the CMC3 and CMC5 treated groups (p>0.05). One patient with B‐CLL and three patients with lymphocytic lymphoma achieved CR (12.1%). Among 12 patients (36.4%) who achieved PR there were 6 CLL patients, and 6 lymphoma patients. The major toxicity was myelosuppression. Severe neutropenia was seen in 11/33 (33.3%) patients, more frequently in patients who received CMC5 than in the patients who received CMC3, both in the CLL (50.0% and 28.5%, respectively) and in the LG‐NHL group (22.2% and 0%, respectively). The rate of thrombocytopenia was similar in both groups. Infections and fever of unknown origin complicated the treatment with CMC5 more often than with CMC3: five episodes were seen in 3 patients treated with CMC3 when compared to 15 episodes in 12 patients treated with CMC5. In conclusion, the CMC programme is an active combined regimen in heavily pre‐treated CLL and LG‐NHL patients. However, its toxicity is significant and we suggest a shortening of 2‐CdA infusion from 5 to 3 d in further studies. Whether a combination of 2‐CdA with cyclophosphamide and mitoxantrone would result in improved outcome as compared to 2‐CdA alone, is being investigated in a prospective, randomised trial.
Stromal‐mediated down‐regulation of CD13 in bone marrow cells originating from acute myeloid leukemia patients
Authors: Dybkær Karen , Olesen Gitte , Skou Pedersen F. , Schøler Kristensen Jørgen
Published: March 2003

Abstract: The metallopeptidase CD13 is expressed on normal myeloid cells of monocytic and granulocytic origin and on the surface of leukemic blasts in most acute myeloid leukemias (AML). To study the mechanisms regulating lineage restricted CD13 expression in AML we determined normalised CD13 mRNA levels in bone marrow cells and peripheral blood cells of 27 AML patients. Cells of bone marrow origin had lower levels of normalised CD13 mRNA than cells of peripheral blood origin, even though fluorescence intensity and fraction of cells expressing CD13 on the surface was unchanged. In particular, AML patients with very low levels of normalised CD13 mRNA in bone marrow cells showed an increase in CD13 mRNA expression in peripheral blood. To evaluate the effects of bone marrow microenvironment on CD13 mRNA expression, we cultured leukemic myeloid cells with and without murine stromal cells. Bone marrow cells with high and low CD13 surface expression that entered the stromal layers all down‐regulated CD13 mRNA expression as compared to cells in suspension above. For peripheral blood cells within stromal layers, CD13 mRNA expression was diminished in only 3 out of 6 cases. The ambiguous effect of stromal cells on peripheral blood cells may illustrate a differentiation‐dependent response towards stroma. We determined the polyadenylation status of CD13 mRNA for 9 bone marrow aspirates and 7 peripheral blood samples. Polyadenylation was diminished in bone marrow cells from AML patients with low levels of normalised CD13 mRNA, raising the possibility of involvement of mRNA instability in regulation of CD13 mRNA expression in this subgroup of patients.
Successful treatment with low‐dose splenic irradiation for massive splenomegaly in an elderly patient with hairy‐cell leukemia
Authors: Nishii Kazuhiro , Katayama Naoyuki , Maeda Hideaki , Usui Eiji , Takemitu Takashi , Lorenzo V Felipe , Nakase Kazunori , Shiku Hiroshi
Published: March 2003

Abstract: The therapeutic approach to hairy‐cell leukemia (HCL) is in some instances still debated. Although management with α‐interferon and purine analogues is well established, there is an alternative role for therapeutic splenectomy in patients with massive splenomegaly who have failed to respond to systemic therapy. Most patients with HCL will not be suitable for treatment with splenectomy as their ages at diagnosis are high. Here, we report an elderly Japanese HCL patient whose refractory massive splenomegaly responded well to low‐dose splenic irradiation.
Decrease of bone marrow angiogenesis in myeloma patients achieving a remission after chemotherapy
Authors: Sezer O. , Niemöller K. , Kaufmann O. , Eucker J. , Jakob C. , Zavrski I. , Possinger K.
Published: March 2003

Abstract: The impact of angiogenesis is well known for the growth and viability of solid tumors. Fewer studies have been published relating angiogenesis to clinical or pathological parameters in hematological malignancies. In this report, we have estimated the bone marrow microvessel density (MVD) before and after conventional‐dose or high‐dose chemotherapy with autologous stem cell transplantation. Immunohistochemical CD34‐stained paraffin‐embedded bone marrow biopsies of 21 patients with stage III multiple myeloma were studied. Microvessels were counted at 400× magnification, and the mean number of vessels per area in each sample was noted as the MVD. The median MVD of all patients was 53.1 vessels/mm2 (range 15.5–174.7 vessels/mm2) before treatment and 29.3 vessels/mm2 (range 0–221.1 vessels/mm2) after chemotherapy. The post‐treatment MVD in the two groups of patients with and without remission was significantly different (p=0.001), whereas the pretreatment MVD was not. Responders but not nonresponders showed a significant decrease of MVD after therapy in comparison to their pretreatment levels. The progression‐free survival in patients who achieved a reduction in MVD after chemotherapy was significantly longer than in patients without a decrease in MVD (P=0.006). Furthermore, we compared the MVD of patients after achievement of a remission to MVD of 15 untreated stage I myeloma patients. The MVD of patients in remission was not statistically different from the MVD in stage I myeloma. These results underscore the impact of angiogenesis in myeloma and give the first report that effective chemotherapy is accompanied by a significant decrease in bone marrow angiogenesis in this disease.
Haematopoietic progenitor cells from the common marmoset as targets of gene transduction by retroviral and adenoviral vectors
Authors: Hibino Hitoshi , Tani Kenzaburo , Sugiyama Hajime , Suzuki Shuzo , Wu Ming‐Shiuan , Izawa Kiyoko , Hase Hidenori , Nakazaki Yukoh , Tanabe Tsuyoshi , Ooi Jun , Izeki Toru , Tojo Arinobu , Saitoh Izumu , Tanioka Yoshikuni , Asano Shigetaka
Published: March 2003

Abstract: To establish a new non‐human primate model for human cytokine and gene therapy, we characterized lymphocytes and haematopoietic progenitor cells of the small New World monkey, the common marmoset. We first assessed the reactions of marmoset bone marrow (BM) and peripheral blood (PB) cells to mouse anti‐human monoclonal antibodies (mAbs) for the purpose of isolating marmoset lymphocytes and haematopoietic progenitor cells. Both cell fractions stained with CD4 and CD8 mAbs were identified as lymphocytes by cell proliferation assay and morphological examination. Myeloid‐specific mAbs such as CD14 and CD33 did not react with marmoset BM and PB cells. No available CD34 and c‐kit mAbs could be used to purify the marmoset haematopoietic progenitor cells.

Furthermore, we studied the in vitro transduction of the bacterial β‐galactosidase (LacZ) gene into CFU‐GM derived from marmoset BM using retroviral and adenoviral vectors. The transduction efficiency was increased by using a mixed culture system consisting of marmoset BM stromal cells and retroviral producer cells. It was also possible to transduce LacZ gene into marmoset haematopoietic progenitor cells with adenoviral vectors as well as retroviral vectors. The percentage of adenovirally transduced LacZ‐positive clusters was 15% at day 4 (multiplicity of infection=200), but only 1–2% at day 14. The differential use of viral vector systems is to be recommended in targeting different diseases. Our results suggested that marmoset BM progenitor cells were available to examine the transduction efficiency of various viral vectors in vitro.
Sustained remission of platelet counts following monoclonal anti‐CD20 antibody therapy in two cases of idiopathic autoimmune thrombocytopenia and neutropenia
Authors: Faurschou Mikkel , Hasselbalch Hans C. , Nielsen Ove J.
Published: March 2003

Abstract: Idiopathic autoimmune thrombocytopenia and neutropenia (ITN) is a primary haemocytopenic disorder clinically characterised by recurrent mucocutaneous bleeding episodes and infections. Unlike in simple idiopathic thrombocytopenic purpura, the platelet deficiency of ITN tends to be chronic and difficult to treat. We describe two patients with ITN who obtained sustained remission of their platelet counts after therapy with the chimeric monoclonal anti‐CD20 antibody Rituximab. In one of two cases, Rituximab also induced prolonged normalisation of the neutrophil count and disappearance of auto‐antibodies.

Our observations indicate that disturbed B‐cell function plays a central role in the pathogenesis of ITN. Anti‐CD20 antibody therapy seems to constitute a safe and efficient alternative to corticosteroids for the management of ITN patients with chronic thrombocytopenia.
Spectral karyotyping and interphase FISH reveal abnormalities not detected by conventional G‐banding
Authors: Nordgren Ann , Heyman Mats , Sahlén Sigrid , Schoumans Jacqueline , Söderhäll Stefan , Nordenskjöld Magnus , Blennow Elisabeth
Published: March 2003

Abstract: Seventy uniformly treated children with acute lymphoblastic leukemia were analysed for chromosomal abnormalities with conventional G‐banding, spectral karyotyping (SKY) and interphase fluorescent in situ hybridisation (FISH) using probes to detect MLL,BCR/ABL, TEL/AML1 rearrangements and INK4 locus deletions. Numerical and/or structural changes could be identified in 80% of the patients by the use of molecular cytogenetic techniques, whereas abnormalities could be detected in 60% of the patients using G‐banding alone. Altogether, 106 structural aberrations were defined by FISH compared to 34 using G‐banding. Seventy‐four percent of the patients had numerical aberrations, 54% structural aberrations and 20% had no identified aberrations. Twelve cases had prognostically unfavourable chromosomal aberrations that had not been detected in the G‐banded analysis. We identified three novel TEL partner breakpoints on 1q41, 8q24 and 21p12, and a recurrent translocation t(1;12)(p32;p13) was found. In addition, two cases displayed amplification (7–15 copies) of AML1. Our results demonstrate the usefulness of SKY and interphase FISH for the identification of novel chromosome aberrations and cytogenetic abnormalities that provide prognostically important information in childhood ALL.
Elevated soluble urokinase plasminogen activator receptor in plasma from patients with idiopathic myelofibrosis or polycythaemia vera
Authors: Jensen Morten Krogh , Riisbro Rikke , De Nully Brown Peter , Brünner Nils , Hasselbalch Hans Carl
Published: March 2003

Abstract: Extracellular proteolytic enzymes of the urokinase‐type plasminogen activator (uPA) system and the family of metalloproteases play a crucial role in the matrix degradation and tissue remodelling processes characteristic of malignant disorders. The receptor for urokinase plasminogen activator (uPAR) serves to localise and intensify the action of uPA and is expressed on the surface of malignant as well as tumour stromal cells including fibroblasts. A soluble form of uPAR (suPAR) cleaved from its glycosyl‐phosphatidylinositol anchor is detected in plasma from healthy individuals and increased levels of suPAR have been found in advanced malignancy, suggesting that suPAR may be a marker of extensive tissue remodelling. In an attempt to clarify whether suPAR might be a marker for bone marrow tissue remodelling we measured plasma suPAR levels in a patient cohort comprising 17 with myelofibrosis (MF), 17 with polycythaemia vera (PV), 15 with essential thrombocythaemia (ET), one with a transitional myeloproliferative disorder evolving from PV and 30 controls. Compared with controls suPAR levels were significantly higher in the patients (P < 0.0001) (median 3.35 vs. 2.32 µg L−1). Moreover, in subgroup analyses including patients with MF, PV, and ET, respectively, suPAR levels differed significantly with the highest levels found in patients with MF and PV (MF vs. PV vs. ET; P = 0.0003). When comparing suPAR levels of the individual patient subgroups with controls, only suPAR levels of PV and MF patients were significantly increased (P < 0.0001). Sixty‐five percent of patients with PV and MF (22/34) had suPAR plasma values that were above the mean +2 standard deviations (SD) of controls. The concentration of suPAR was significantly correlated to plasma lactate dehydrogenase, thrombomodulin, and complex of tPA:PAI‐1 in the patients. There was no difference between patients and controls when comparing plasma uPA levels. Increased plasma suPAR levels in patients with chronic myeloproliferative disorders may reflect increased uPAR production in the bone marrow, leading to enhanced bone marrow remodelling.
Molecular determinants of UV‐induced immunosuppression
Authors: Schwarz Agatha , Schwarz Thomas
Published: March 2003

Abstract: It is almost three decades ago that it was discovered that ultraviolet radiation (UV) can compromise the immune system. UV suppresses immune responses in several ways. It inhibits the function of antigen‐presenting cells, induces T cells with suppressor activity and induces the release of immunosuppressive cytokines. The latter phenomenon is mainly responsible for systemic immunosuppression. Although UV can also target cytoplasmic and cell membrane components, UV‐induced DNA damage has been recognized as the most important molecular structure in mediating UV‐induced immunosuppression. Recently, it was observed that interleukin‐12 (IL‐12), which antagonizes UV‐induced immunosuppression, can accelerate the removal of UV‐induced DNA lesions, probably via inducing DNA repair. Hence, it is tempting to speculate that the activity of IL‐12 to reduce UV‐induced immunosuppression may be due at least partially to this new biological activity of IL‐12.
Protection against UV‐induced suppression of contact hypersensitivity responses by sunscreens in humans
Authors: Cooper K. D. , Baron E. D. , LeVee G. , Stevens S. R.
Published: March 2003

Abstract: Both in vivo skin immune responses and the skin's reaction to sun exposure integrate a complex interplay of biologic responses. The complexity and multiplicity of events that occur in the skin during an immune response make it a sensitive indication of both UVB and UVA‐induced changes in the skin by sun damage, as well as those changes that are prevented by various sunscreens. Sunscreens are the most effective and widely available intervention for sun damage, other than sun avoidance or clothing. However, sunscreens vary widely in their relative ability to screen various UV waveband components, and their testing has been variably applied to outcomes other than for erythema to determine the sunburn protection factor (SPF), a measure primarily of UVB filtration only. Determination of an immune protection factor (IPF) has been proposed as an alternative or adjunctive measure to SPF, and recent studies show IPF can indeed detect added in vivo functionality of sunscreens, such as high levels of UVA protection, that SPF cannot. Clarification of the definition of IPF, however, is required. Excellent data are available on quantification of the IPF for restoring the afferent or induction arm of contact sensitivity, but other immune parameters have also been measured. Proposed here is nomenclature for whether the IPF is measured using contact sensitivity induction (IPF‐CS‐I), contact sensitivity elicitation (IPF‐CS‐E), delayed‐type hypersensitivity elicitation (IPF‐DTH‐E), antigen‐presenting cell function (IPF‐APC‐FXN) or numbers (IPF‐APC‐#), and cytokine modification such as IL‐10 (i.e. IPF‐cyto‐IL‐10). Similar nomenclatures could be used for other measures of skin function protection (i.e. DNA damage, p53 induction, oxidation products, etc.). A review of in vivo human studies, in which sunscreens are used to intervene in a UV‐induced modulation of immune response, cells or cytokines, highlights the technical variables and statistical approaches which must also be standardized in the context of an IPF for regulatory or product claim purposes. Development of such IPF standards would allow the integration of both UVB and nonUVB (UVA, blue and possible IR) solar waveband effect‐reversals, could be applied to integrate effects of other ingredients with protective function (i.e. antioxidants, retinoids, or other novel products), and would spur development of more advanced and complete protection products.
Transgene expression in human epidermal keratinocytes: cell cycle arrest of productively transfected cells
Authors: Jensen U. B. , Petersen M. S. , Lund T. B. , Jensen T. G. , Bolund L.
Published: March 2003

Abstract: We have analysed the consequences of liposome mediated gene transfer into human primary epidermal keratinocytes and compared non‐Epstein–Barr Virus (EBV) and EBV based expression vectors that carry the genes encoding human Growth Hormone (hGH) or Enhanced Green Fluorescent Protein (EGFP). Different kinetics between the non‐EBV and EBV based vectors were revealed upon subcultivation of hGH transfected keratinocytes. The keratinocytes transfected with non‐EBV based vector showed a rapid reduction in hGH production. Although the EBV based vector resulted in more stable expression, this was also reduced over time. Chromatin inactivation by deacetylation was investigated by treatment with sodium butyrate and found not to be the reason for the decreasing expression. Keratinocytes divided into subpopulations enriched for either stem cells or transit amplifying cells, based on β1‐integrin expression and function, do not differ significantly with respect to susceptibility to productive transfection. However, when the keratinocytes were transfected with the EGFP gene and sorted live by FACS into EGFP negative and positive populations, only the negative cells were capable of forming significant numbers of colonies. This is consistent with the observation that the ability to incorporate BrdU was dramatically reduced in the EGFP expressing population within 24–48 h post transfection indicating an almost complete cell cycle arrest. p53 levels were unaffected by the procedures, and the keratinocyte cell line HaCat, mutated in both p53 alleles, also shows a marked reduction in clonogenic potency upon transfection. There was a slight increase of TUNEL positive apoptotic nuclei in the positive population at early time points. However, the apoptotic index was still very low. When we measured the frequency of involucrin expressing cells, we found an increase in the productively transfected population over time indicating an initiation of terminal differentiation. In contrast to the transfected cultures, keratinocytes that were transduced using a retroviral vector showed no decrease in colony forming efficiency. In conclusion we find that transgene expressing cells from transfected cultures of epidermal keratinocytes undergo cell cycle arrest and initiate terminal differentiation by mechanisms which are independent of p53 levels.
RNA as a tumor vaccine: a review of the literature
Authors: Saenz‐Badillos J. , Amin S. P. , Granstein R. D.
Published: March 2003

Abstract: Many approaches have been attempted to harness the host immune system to act against malignant tumors. These have included animal and clinical trials with agents to non‐specifically boost immunity, factors to augment specific immunity, transfer of lymphokine‐activated killer cells and transfer of expanded populations of tumor‐infiltrating lymphocytes. Therapeutic vaccination strategies have been employed using tumor extracts, purified tumor antigens, recombinant peptide tumor antigens and specific DNA sequences coding for a tumor antigen (genetic vaccination) both through direct administration to the host and by administration of antigen presenting cells exposed to these materials ex vivo. Recently, the use of RNA has been proposed for use in tumor vaccination protocols. The use of RNA has several potential advantages. Since total cellular RNA or mRNA can be utilized, it is not necessary to know the molecular nature of the putative tumor antigen(s). RNA can be effectively amplified; thus, unlike tumor‐extract vaccines, only a small amount of tumor is needed to prepare the material for vaccination. Also, unlike DNA‐based vaccines, there is little danger of incorporation of RNA sequences into the host genome. The possible utility of RNA‐based vaccines for tumor immunotherapy should be further explored to determine whether such approaches are clinically useful.
Identification of immature cornified envelopes in the barrier‐impaired epidermis by characterization of their hydrophobicity and antigenicities of the components
Authors: Hirao T , Denda M. , Takahashi M.
Published: March 2003

Abstract: Cornified envelopes (CEs), rigid and insoluble structures in the stratum corneum, which are assembled by crosslinking of several precursor proteins by transglutaminases, provide a hydrophobic foundation for barrier function; omega‐hydroxyceramides are covalently attached to the outer surface of CE components, and onto this hydrophobic assembly, lamellar layers of intercellular lipids are organized. Morphologically irregular, fragile CEs are found in the deep layer of the stratum corneum or in certain disorders, such as psoriasis, whereas most CEs from healthy subjects are rigid and polygonal. We have established a staining method to characterize such fragile CEs as immature and less hydrophobic CEs, and employed it to examine regional differences in the properties of CEs, especially in relation to the barrier function of the skin. CEs from the outermost stratum corneum of the trunk and extremities of healthy subjects were relatively uniform in morphology with larger shape, and were homogeneous in hydrophobicity as judged from the use of an environment‐sensitive fluorescent dye, Nile red. However, CEs from the face were strikingly heterogeneous, and consisted of both rigid and fragile CEs. Rigid CEs were Nile red‐positive and little stained by anti‐involucrin. In contrast, fragile CEs were Nile red‐negative but strongly stained with anti‐involucrin, as detected by indirect immunofluorescence. Thus, CEs from the face were stained with Nile red or involucrin in a mutually exclusive manner. Fragile CEs were stained with antibodies against other CE components, including loricrin, envoplakin, filaggrin, and isopeptides. Such fragile, involucrin‐positive CEs were detected not only in the face, but also in the deep layer of the stratum corneum of the arm. In addition, experimental barrier disruption resulted in the appearance of involucrin‐positive CEs in the outermost stratum corneum. These results suggest that involucrin‐positive, fragile CEs are immature and less hydrophobic, and that their occurrence is closely related to impairment of the barrier function of the skin
Psoriasis scales contain C5a as the predominant chemotaxin for monocyte‐derived dendritic cells
Authors: Mrowietz U. , Koch W.‐A. , Zhu K. , Wiedow O. , Bartels J. , Christophers E. , Schröder J.‐M.
Published: March 2003

Abstract: Dendritic cells seem to be of major importance for the pathogenesis of psoriasis. They are increased in number in lesional psoriatic skin which is thought to be due to an increased influx from the peripheral blood regulated by chemotaxins. Using a biological/biochemical approach we have addressed the question whether psoriasis scale extracts contain proteinaceous chemotaxins for dendritic cells. Human monocytes differentiated into dendritic cells by culture with GM‐CSF and IL‐4 (MoDC) served as responder cells. Chemotactic activity for MoDC was purified by several HPLC‐steps. The results of our study show that C5a/C5adesarg is the major chemotactic peptide for MoDC in psoriasis scale extracts. In comparison to other stimuli such as fMLP or monocyte chemotactic peptide 1 (MCP‐1) C5a proved to be a most potent and efficient chemotaxin for MoDC. C5a co‐eluted with MRP14/calgranulin B which is present in large amounts in psoriasis scale extracts as identified by amino acid sequencing. However, MRP14/calgranulin B did not possess any chemotactic activity for MoDC. Our results provide evidence that C5a/C5adesarg although not specific for dendritic cells seems to be the major chemoattractant for these cells in lesional psoriasis skin.