bluuebunny/crossref_metadata_2025 huggingface dataset:
High proportions of VLA‐5− immature myeloma cells correlated well with poor response to treatment in multiple myeloma
Authors: Kawano Michio M. , Mahmoud Maged S. , Huang Naihui , Lisukov Igor A. , Mihara Keiichiro , Tsujimoto Takako , Kuramoto Atsushi
Published: March 2008

Summary. Using two‐colour phenotypic analysis with anti‐CD38 antibody, human myeloma cells can be classified into VLA‐5− immature and VLA‐5+ mature cells. We examined the relationship between variations of these subpopulations and clinical responses during treatment in multiple myeloma (MM). 39 patients with MM were treated with combined chemotherapy. First estimation of clinical responses after induction therapy showed that early clinical responses were correlated with the percentage of immature myeloma cells present after induction therapy (P < 0.01), not at diagnosis. After three courses of cyclic maintenance therapy, immature myeloma cells significantly decreased in proportion along with a decrease in total myeloma cells in maintained or more responsive cases (P < 0.01). On the other hand, immature myeloma cells were still found in high proportions in non‐responsive cases with no change (NC) or minor response (MR) (P < 0.01). Furthermore, in relapsing cases from partial response (PR) or progressive disease (PD) from non‐responsive cases, immature myeloma cells increased markedly. Therefore these results show that high proportions of VLA‐5− immature myeloma cells remaining after induction therapy and during maintenance therapy correlate well with a declining clinical course of MM during maintenance therapy.
Disappearance of AML1 ‐MTG8(ETO) fusion transcript in acute myeloid leukaemia patients with t(8;21) in long‐term remission
Authors: Satake Noriko , Maseki Nobuo , Kozo Tomoko , Sakashita Akiko , Kobayashi Hirofumi , Sakurai Masaharu , Ohki Misao , Kaneko Yasuhiko
Published: March 2008

Summary. In a study of 23 patients with t(8;21)‐associated acute myeloid leukaemia the AML1‐MTG8 fusion transcript was present in the majority of serial samples obtained from 17 patients followed for up to 34 months after diagnosis, but was absent in samples from all six patients who had been in continuous complete remission for 61 months after allogeneic bone marrow transplantation (BMT), or for 52, 53, 123,182 and 198 months, respectively, after courses of intensive chemotherapy. Previous studies showed that the AML1‐MTG8 fusion transcript was present in most patients with this type of translocation in long‐term remission. Our results indicate that blood cells of patients with t(8;21) in remission of over 10 years may not show the AML1‐MTG8 fusion transcript, and that those of patients who have undergone allogeneic BMT or intensive chemotherapy may become fusion transcript‐negative much earlier. Our study suggests that leukaemic cells with the AML1‐MTG8 fusion transcript may survive for some time after courses of chemotherapy or BMT, but that they may eventually be eradicated by immunologic and other antileukaemic mechanisms.
Peripheral primitive neuroectodermal tumour after allogeneic bone marrow transplantation
Authors: Gamelin Erick , Francois Sylvie , Guyetant Serge , Rousselet Marie Christine , du Chatelard Pierre Pabot , Coindre Jean Michel , Ifrah Norbert
Published: March 2008

Summary. We report a case of peripheral primitive neuroectodermal tumour (PNET) in a 36‐year‐old man who 10 years earlier received an allogeneic bone marrow transplantation (BMT) for a non‐Hodgkin's lymphoma. Immunohistochemistry proved definitive elimination of a relapse of the original disease and confirmed the diagnosis of PNET. The role of total body irradiation pre‐BMT in the genesis of this secondary tumour is discussed. The importance of registering secondary malignant solid tumours after BMT is emphasized. To our knowledge, an Ewing family tumour following BMT has not previously been reported.
EBV‐associated lymphoproliferative syndrome with a distinct 69 base‐pair deletion in the LMP‐1 oncogene
Authors: Klein Christoph , Rothenberger Sylvia , Niemeyer Charlotte , Bachmann Edith , Odermatt Bernhard , Bohm Norbert , Brandis Matthias , Knecht Hans
Published: March 2008

Summary. We describe an immunocompetent 12‐year‐old boy with chronic EBV infection and lymphoid interstitial pneumonitis. Lymph node biopsies showed effacement of the architecture with polymorphic cellular infiltrates, consisting predominantly of T cells and natural killer cells. No clonal rearrangement of TCR or immunoglobulin genes was seen. DNA was extracted from hilar lymph nodes; sequencing of the carboxy terminal region of the latent membrane protein 1 (LMP‐1) oncogene revealed a 69 base‐pair deletion and four point mutations. Immunosuppressive treatment with prednisone and cyclosporine reversed the lymphadenopathy.
Incidence of human parvovirus B19 DNA detection in blood donors
Authors: Yoto Yuko , Kudoh Tooru , Haseyama Keiji , Suzuki Nobuhiro , Oda Takanori , Katoh Toshiaki , Takahashi Tsuneo , Sekiguchi Sadami , Chiba Shunzo
Published: March 2008

Summary. 1000 serum samples from blood donors were tested for human parvovirus B19 (B19) DNA by a nested PCR assay: six samples were positive for B19 DNA. The frequency was 1/167 (0‐6%), considerably higher than previous surveys (0‐004‐0‐03%). Five of the six samples were also positive for anti‐B19 IgM, indicating an acute phase of infection. It is recommended to screen for B19 DNA in blood products to prevent transfusion mediated viral infection for those susceptible such as immunocompromised patients and pregnant women.
Expression of erythroid‐specific genes in acute megakaryoblastic leukaemia and transient myeloproliferative disorder in Down's syndrome
Authors: ITO ETSURO , KASAI MIKIO , HAYASHI YASOHIDE , TOKI TSUTOMU , ARAI KOJI , YOKOYAMA SHINKICHI , KATO KOJI , TACHIBANA NAOKI , YAMAMOTO MASAYUKI , YOKOYAMA MASARU
Published: March 2008

Summary. Acute megakaryoblastic leukaemia (M7) and transient myeloproliferative disorder in Down's syndrome (TMD) are characterized by rapid growth of abnormal blast cells which express megakaryocytic markers. To clarify properties of the blast cells in M7 and TMD cases, we examined erythroid markers expression in blasts from six cases with M7 and seven cases with TMD in this study. Erythroid‐specific mRNAs encoding 7‐globin and erythroid 6‐aminolevulinate synthase were found to be expressed in blasts from most of these cases, indicating that majorities of the blasts in M7 and TMD cases have erythroid and megakaryocytic phenotypes. We also found that mRNAs encoding GATA‐1 and GATA‐2 are expressed in all these cases. These results suggest that M7 blasts and TMD blasts correspond to the erythroid/megakaryocytic bipotential progenitor cells.
Maternal alloimmunization against fetal platelet antigens: a prospective study
Authors: PANZER SIMON , AUERBACH LEO , CECHOVA EVA , FISCHER GOTTFRIED , HOLENSTEINER ANDREA , KITTL EVA‐MARIA , MAYR WOLFGANG RICHARD , POTZ MICHAEL , WAGENBICHLER PETER , WALCHSHOFER SABINE
Published: March 2008

Summary. Neonatal alloimmune thrombocytopenia (NAIT) is induced by maternal alloantibodies to fetal platelet antigens. This prospective study was carried out to evaluate the incidence of anti‐platelet antibodies in 933 mother‐child pairs where the mother and child were typed for the human platelet antigens (HPA)‐l, ‐2,‐3,‐5. Sera from mismatched mother‐child pairs were screened for anti‐platelet antibodies, anti‐HLA class I and blood group ABO IgG antibodies. Platelet‐specific antibodies were anti‐HPA‐3a in one and anti‐HP A‐5b in 17 neonates, respectively. All these neonates had normal platelet counts. One woman had autoreactive antibodies. Anti‐HLA class I and anti‐blood group A IgG antibodies were detected in five and four neonates, respectively, born with a platelet count <150×109/l. None of the 11 homozygous HP A‐lb mothers became immunized against their heterozygous offspring. The maternal HLA‐allotypes HLA‐DR52 and ‐DR6, typically found in individuals immunized against HPA‐la and ‐5b, respectively, were found in three of 11 HPA‐b/b non‐responders and eight of the anti‐HPA‐5b responders. The results indicate that a risk for NAIT due to HPA‐2 and ‐3 alloimmunization is low. The HLA allotypes do not predict the risk for NAIT due to HPA‐1 or ‐5 alloimmunization. Maternal anti‐HPA‐5b antibodies do not correlate with the platelet count in the neonate.
Development of resistance to activated protein C during pregnancy
Authors: CUMMING A. M. , TAIT R. C. , FILDES S. , YOONG A. , KEENEY S. , HAY C. R. M.
Published: March 2008

Summary. We measured activated protein C (APC) anticoagulant activity in 20 healthy women at 14‐20, 28 and 36 weeks gestation, and at 1 d post‐partum. Significant reductions in the mean APC sensitivity ratio (APC‐SR) were observed at all stages of pregnancy studied compared with the mean APC‐SR obtained for baseline measurements carried out at > 8 weeks post‐partum. APC resistance was seen in 8/19 (42%) and in 11/20 (55%) women at 14‐20 and 28 weeks gestation respectively. The development of resistance to APC may contribute to the increased risk of thrombosis during pregnancy.
Retinoic acid inhibits the growth of human myeloma cells in vitro
Authors: Palumbo Antonio , Battaglio Silvano , Napoli Patrizia , Bruno Benedetto , Omedè Paola , Boccadoro Mario , Pileri Alessandro
Published: March 2008

Retinoic acid has been shown to induce growth inhibition in a variety of cell types including human myeloma cell lines. Bone marrow plasma cells from 31 multiple myeloma (MM) patients were cultured to investigate the activity of 13‐cis‐retinoic acid (cRA), all‐trans‐retinoic acid (tRA), interferon‐α (IFN‐α), interferon‐γ (IFN‐γ), and dexamethasone (DEX), alone or in combination, on in vitro proliferation and immunoglobulin (Ig) secretion. Both cRA and tRA inhibited proliferation: the labelling index (LI) of treated cultures/controls, was 0.47±0.05 (mean ± standard error mean, M ± SEM) P < 0.0001, and 0.67 ± 0.04 (M ± SEM), P < 0.0001, respectively. The inhibitory effect of cRA was significantly superior to tRA (P= 0.0129) and IFN‐α, similar to IFN‐γ and DEX. The combinations of cRA + IFNα, tRA + IFN‐γ, tRA + DEX did not show any synergistic effect on myeloma proliferation. In contrast, the combination cRA + DEX (0.29 ± 0.04, M±SEM) markedly increased the effect of both cRA and DEX used as single agents.

Ig synthesis was not significantly affected by CRA, tRA, IFN‐γ and the combination tRA + IFN‐γ, As expected, only IFN‐α (P 0.002) and DEX (P 0.001) inhibited Ig production. The combinations cRA + IFN‐α, cRA‐DEX and tRA + DEX decreased Ig secretion to the same extent as IFN‐α and DEX alone respectively. In conclusion, our data indicate that tRA and especially cRA strongly inhibited plasma cell proliferation but had no effect on Ig synthesis. The combination of cRA + DEX showed the highest degree of inhibitory activity of all cytokines. alone or in combination.
Thrombotic thrombocytopenic purpura‐like syndrome in the absence of schistocytes
Authors: Fava Stephen , Galizia Alfred Caruana
Published: March 2008

Thrombotic thrombocytopenic purpura is anuncommon disorder that if left untreated has a very high mortality. Schistocytes are generally considered essential for the diagnosis. A patient is presented with a thrombotic thrombocytopenic purpura‐like syndrome in whom schistocytes were persistently absent and who responded to plasmapheresis.
Transformation into acute basophilic leukaemia in a patient with myelodysplastic syndrome
Authors: Yamagata Tetsuya , Miwa Akiyoshi , Eguchi Mitsuoki , Kitagawa Seiichi , Muroi Kazuo , Hatake Kiyohiko , Suda Toshio , Sakamoto Shinobu , Miura Yasusada
Published: March 2008

We describe a patient with basophilic leukaemia following a 2‐year period with myelodysplastic syndrome (refractory anaemia). The marrow showed 59.4% of blasts with 25.0% of mature and immature basophils. The leukaemic blasts contained granules, positively stained with toluidine blue but negative for peroxidase. The basophilic differentiation was confirmed by ultrastructural analysis demonstrating immature basophil granules. In addition, a morphological transition from immature blasts to more mature basophils was observed. Immunophenotypic analysis of blasts and basophils showed positive for CD5, CD7, CD13, CD33 and CD34. Cytogenetic investigation showed an abnormal karyotype, 46,XY,del(5)(q31q35). in 11% of the cells examined when the initial diagnosis of refractory anaemia was made. However, expansion of the same clone up to 100% was observed concomitantly with transformation to basophilic leukaemia.
Persistent lymphocytosis of natural killer cells in autoimmune thrombocytopenic purpura (ATP) patients after splenectomy
Authors: Garcia‐Suarez J. , Prieto A. , Reyes E. , Arribalzaga K. , Perez‐Machado M. A. , Lopez‐Rubio M. , Manzano L. , Alvarez‐Mon M.
Published: March 2008

We report three patients with primary autoimmune thrombocytopenic purpura (ATP) who developed an absolute lymphocytosis (lymphocyte count > 5×109/I) after splenectomy and with a lymphocyte count between 5.4 and 8.9×109/I. An immunophenotype study showed that the peripheral blood lymphocytosis was a persistent NK cell expansion (CD2+, CD56+, CD3‐), and was characterized by a typical large granular lymphocytes (LGL) morphology. Two of these three ATP patients were refractory to splenectomy.
A novel candidate mutation (Arg611→ His) in type I‘platelet discordant’von Willebrand's disease with desmopressin‐induced thrombocytopenia
Authors: Castaman Giancarlo , Eikenboom Jeroen C. J. , Rodeghiero Francesco , Briët Ernest , Reitsma Pieter H.
Published: March 2008

In three affected members of a family with type I platelet discordant’von Willebrand's disease displaying desmopressin‐induced thrombocytopenia, we have detected in exon 28 of the von Willebrand factor gene a heterozygous G(4121) → A transition, which predicts an Arg611→ His substitution. The mutated allele was absent in 50 normal individuals. An unrelated patient with a similar phenotype was also found to be heterozygous for this mutation. The mutation is located in the A1 domain of von Willebrand factor, where most type 2B von Willebrand's disease mutations are found. Mutations in this domain result in von Willebrand factor multimers with enhanced affinity for platelet glycoprotein Ib, and this may explain the association of Arg611→ His with the moderate thrombocytopenia observed after desmopressin infusion.
Co‐segregation of thrombosis with the factor V Q506 mutation in an extended family with resistance to activated protein C
Authors: Beufé Sylvie , Borg Jeanne‐Yvonne , Vasse Marc , Charbonnier Francloise , Moreau Viviane , Monconduit Mathieu , Frebourg Thierry
Published: March 2008

The activated protein C (APC) resistance phenotype results from a mutation at one of the cleavage sites of factor V by APC (Q506). We describe a large family with an APC resistance phenotype and without any other detectable coagulation defect, including eight subjects who had developed deep venous thrombosis (mean age of the first thrombosis episode 29 years; range 17‐55 years). The factor V Q506 mutation was detected in the seven patients with thrombosis who could be tested and in 13 asymptomatic subjects (mean age 17 years; range 5‐33 years). The APC resistance was detectable in only 10 heterozygotes among the 19 tested. These data suggest that, in affected families, the risk for the factor V Q506 mutation carriers to develop thrombosis may be very high and that factor V genotyping must be performed in patients with thrombosis even without any detectable APC resistance phenotype.
Pollen beetle in the UK; the start of a resistance problem?
Authors: Richardson D. M.
Published: March 2008

In 2003, the first report of poor control of pollen beetle Meligethes aeneus at a site in South East England in the UK was investigated but resistance to pyrethroid insecticides was not confirmed in subsequent laboratory testing. Bioassays of 26 UK samples of M. aeneus collected in 2004 with the pyrethroid lambda‐cyhalothrin showed little or no divergence from the response expected of a fully susceptible strain. In 2006 samples of pollen beetle from the UK were sent to Germany, and again these were shown to be fully susceptible. In 2007 using test kits supplied by Udo Heimbach, BBA, 19 samples of pollen beetle were tested, again from across the UK. Results indicated that a small number of individuals were fully resistant, surviving the highest dose of lambda‐cyhalothrin tested (0.375 microg/L) after 5‐h exposure at 4 sites, and after 24‐h exposure at 2 of these sites.
Thrips hawaiiensis (Morgan, 1913) (Thysanoptera: Thripidae), an Asian pest thrips now established in Europe
Authors: Reynaud P. , Balmès V. , Pizzol J.
Published: March 2008

An established population of Thrips hawaiiensis (Morgan) (Thysanoptera: Thripidae), a species originating from Asia and the Pacific, is reported for the first time in the Western Palaeartic zone (in the South of France). Notes relating to the current geographical distribution, identification, biology and risk of establishment of this potentially invasive species in Europe are provided.
Reconciling poverty alleviation with reduction in fisheries capacity: boat aid in post‐tsunami Aceh, Indonesia
Authors: TEWFIK A. , ANDREW N. L. , BENE C. , GARCES L.
Published: March 2008

Abstract  The tremendous loss of life and assets resulting from the 2004 tsunami dealt a devastating blow to the coastal communities of Aceh Province, Indonesia. An assessment of the fishing fleet structure pre‐ and post‐tsunami, including associated pattern in boat aid, in 15 coastal communities was conducted and compared with data on boat relief efforts over 17 districts of the Province. Aid was found to be not proportionally allocated to losses incurred by communities and was in many cases below what could be seen as a trend toward overcapacity. The distribution of aid appeared to be done without consideration of the former structure of the fleet, which changed significantly over time (pre‐ and post‐tsunami), and resulted in a new fleet of reduced diversity with a strong focus on the smaller boat categories. If this situation perpetuates after the initial post‐disaster period, it may further exacerbate the pre‐existing economic and geographic marginalisation of some remote communities, and reduce the capacities of the fleets to diversify and develop multi‐species harvesting strategies; thus, increasing the risk of ecologically unsustainable exploitation in near shore areas.
Testing Bergmann’s rule in the presence of potentially confounding factors: a case study with three species of Galerida larks in Morocco
Authors: Guillaumet Alban , Ferdy Jean‐Baptiste , Desmarais Eric , Godelle Bernard , Crochet Pierre‐André
Published: March 2008

Aim  To test Bergmann’s rule (which predicts a larger body size in colder areas within warm‐blooded vertebrate species) in three partially sympatric species of larks (Galerida theklae, Galerida cristata and Galerida randonii) that occur in Morocco.

Location  Morocco.

Methods  Restriction fragment length polymorphism techniques applied on cytochrome b haplotypes were used to discriminate G. cristata and G. randonii, and to investigate the effects of interspecific hybridization in their contact zone. A comprehensive statistical framework was then designed to test Bergmann’s rule in our three Galerida species (using altitude as a proxy for cold temperatures), while controlling for the possible influence of interspecific hybridization and competition and accounting for spatial autocorrelation. The method we propose is conservative in the sense that potentially confounding factors are adjusted so as to maximize their influence on the variable of interest.

Results  Bergmann’s rule was strongly supported in G. theklae and G. randonii. However, body size did not respond to altitude in G. cristata, a result that was not simply explained by species‐specific differences in geographical ranges and altitudinal span. In G. cristata, we detected a tendency for body size to increase with aridity, in agreement with an alternative definition of Bergmann’s rule. However, since G. cristata also hybridizes with G. randonii in a contact zone located in the most arid part of the range of G. cristata, we could not tease apart the relative contribution of selection and hybridization in driving this pattern.

Main conclusions  This study highlights the need for careful statistical designs that allow meaningful variables to be picked out from large sets of potential factors. When taking these factors into account, we found that Bergmann’s rule was still strongly supported in two out of the three species examined.
Mesophyll conductance to CO2: current knowledge and future prospects
Authors: FLEXAS JAUME , RIBAS‐CARBÓ MIQUEL , DIAZ‐ESPEJO ANTONIO , GALMÉS JERONI , MEDRANO HIPÓLITO
Published: March 2008

During photosynthesis, CO2 moves from the atmosphere (Ca) surrounding the leaf to the sub‐stomatal internal cavities (Ci) through stomata, and from there to the site of carboxylation inside the chloroplast stroma (Cc) through the leaf mesophyll. The latter CO2 diffusion component is called mesophyll conductance (gm), and can be divided in at least three components, that is, conductance through intercellular air spaces (gias), through cell wall (gw) and through the liquid phase inside cells (gliq). A large body of evidence has accumulated in the past two decades indicating that gm is sufficiently small as to significantly decrease Cc relative to Ci, therefore limiting photosynthesis. Moreover, gm is not constant, and it changes among species and in response to environmental factors. In addition, there is now evidence that gliq and, in some cases, gw, are the main determinants of gm. Mesophyll conductance is very dynamic, changing in response to environmental variables as rapid or even faster than stomatal conductance (i.e. within seconds to minutes). A revision of current knowledge on gm is presented. Firstly, a historical perspective is given, highlighting the founding works and methods, followed by a re‐examination of the range of variation of gm among plant species and functional groups, and a revision of the responses of gm to different external (biotic and abiotic) and internal (developmental, structural and metabolic) factors. The possible physiological bases for gm, including aquaporins and carbonic anhydrases, are discussed. Possible ecological implications for variable gm are indicated, and the errors induced by neglecting gm when interpreting photosynthesis and carbon isotope discrimination models are highlighted. Finally, a series of research priorities for the near future are proposed.
Differentiation of Veillonella atypica, Veillonella dispar and Veillonella parvula using restricted fragment‐length polymorphism analysis of 16S rDNA amplified by polymerase chain reaction
Authors: Sato T. , Matsuyama J. , Sato M. , Hoshino E.
Published: March 2008

Veillonella atypica, Veillonella dispar and Veillonella parvula cannot be reliably distinguished by conventional phenotypic tests, including the API ZYM test. In this study, restricted fragment‐length polymorphism (RFLP) analysis of 16S rDNA amplified by polymerase chain reaction (PCR) was used to generate restriction profiles of the type strains of V. atypica, V. dispar and V. parvula and 20 Veillonella strains isolated from oral sites. 16S rRNA gene sequences from isolated genomic DNA samples were amplified by PCR. PCR products were purified and characterized by single digestion with 13 restriction endonucleases. Among them, MnlI was found to discriminate the respective reference strains, and the clinical isolates were assigned to one of the three species on the basis of their restriction profiles by digestion with MnlI. Thus, RFLP analysis of PCR‐amplified 16S rDNA, using MnlI, is a rapid and reliable method for the differentiation of V. atypica, V. dispar and V. parvula.
Phenotypic and genotypic characterization of oral yeasts from Finland and the United States
Authors: Hannula J. , Saarela M. , Alaluusua S. , Slots J. , Aslkainen S.
Published: March 2008

A total of 4–22 isolates of oral yeasts per subjects from 48 yeast‐positive Finnish and American subjects (25 females and 23 males) were phenotyped and genotyped to determine the frequency of simultaneous oral carriage of multiple yeast taxa. An oral sample from either periodontal pockets, oral mucosa or saliva was obtained. All subjects yielded Candida albicans and 3 subjects an additional yeast species (Candida krusei, Candida glabrata or Saccharomyces cerevisiae). The API 20C Aux kit distinguished 9 different carbohydrate assimilation profiles among the C. albicans isolates. Thirty‐eight of 46 C. albicans biotype I isolates were categorized in a single numerical profile. PCR analysis, using a random primer OPA‐03 and a repetitive primer (GACA)4, detected 2 major genotypic groups among the C. albicans isolates; 44 subjects showing isolates with a “typical” PCR‐profile and 4 subjects isolates with an “atypical” PCR‐profile. The “atypical” PCR‐profile was similar to that of Candida dubliniensis. All C. albicans isolates assimilated xylose, except 5, including the 4 with an “atypical” PCR‐profile. No difference was found in distribution of oral yeast species, and of C. albicans phenotypes and genotypes between Finnish and American subjects. The present PCR method may offer a rapid and easy means of distinguishing oral Candida species.
In vitro antimicrobial susceptibility of periodontopathic Actinobacillus actinomycetemcomitans to roxithromycin and erythromycin
Authors: Plccolomini R. , Bonaventura G. Di , Catamo G. , Picciani C. , Paolantonio M.
Published: March 2008

The in vitro minimal inhibitory concentrations (MIC) and minimal bactericidal concentration (MBC) of roxithromycin and erythromycin against Actinobacillus actinomycetemcomitans were evaluated. Sixty‐seven different A. actinomycetemcomitans isolated from periodontal pockets of 101 subjects with different forms of early‐onset and adult periodontitis and three reference strains of A. actinomycetemcomitans (ATCC 29522, ATCC 29523, and NCTC 9710) were included in this study. Erythromycin showed poor in vitro activity against A. actinomycetemcomitans; roxithromycin, on the contrary, exhibited good in vitro activity. Moreover, roxithromycin showed the best in vitro antimicrobial activity against 17 serotype a and 12 serotype c subpopulations of A. actinomycetemcomitans; against 38 serotype b subpopulation of A. actinomycetemcomitans, roxithromycin was consistently active. Roxithromycin exhibited MBC values usually equal to, or onefold higher than MIC values. All the MBC values of erythromycin were three‐to four‐fold higher than the respective MIC result. Since roxithromycin is characterized by high concentrations in serum and good penetration and diffusion into gingival tissue, it could be expected to pass into the gingival crevicular fluid at levels sufficiently high to inhibit A. actinomycetemcomitans in vivo. These data indicate that roxithromycin might be a potential candidate for therapeutic trials in patients with A. actinomycetemcomitans‐associated periodontitis.
Differential activation of human neutrophils by Streptococcus mutans isolates from root surface lesions and caries‐free and caries‐active subjects
Authors: Moore M. A. , Gregory R. L. , Switalski L. M. , Hakki Z. W. , Gfell L. E. , Kowollk M. J.
Published: March 2008

Phagocytosis of bacterial pathogens is an important defense mechanism and may contribute to regulating Streptococcus mutatis‐induced dental caries, particularly at root surfaces. This study was undertaken to examine and compare differences in polymorphonuclear leukocyte or neutrophil activation by clinical isolates of S. mutans collected from the saliva of caries‐free or caries‐active individuals with S. mutans isolates from root surface lesions. S. mutans clinical isolates (5 caries‐free, 5 caries‐active, 5 root caries isolates and a laboratory strain) were incubated with neutrophils in the presence of normal human serum and the luminol dependent chemiluminescence was measured for 1 h at 37°C. Results indicated that the caries active and laboratory strains activated neutrophils equally. The mean integration stimulated by caries‐free strains, however, displayed a 25–30% enhanced neutrophil activation over the caries‐active and laboratory strains. In contrast, neutrophil activation by root caries strains of S. mutans was 45–50% lower than all other S. mutans strains, possibly suggesting a natural selection for S. mutans strains that can evade neutrophil recognition and subsequent phagocytosis. Stimulation of neutrophils with the cell wall and membrane surface component preparations indicated that extracts from all four groups activated neutrophils significantly. Again, caries‐free preparations activated neutrophils significantly more than caries active, laboratory strain and root caries isolates. This selection may become more important on root surfaces due to increased exposure to crevicular fluid and neutrophils. The data provide evidence for the presence or onset of mechanisms or biological alterations in S. mutans developed to circumvent neutrophil recognition and/or phagocytosis, thus increasing S. mutans survival and colonization on tooth surfaces, resulting in an enhanced risk of dental caries, particularly at root surfaces.
Cystatin D, a natural salivary cysteine protease inhibitor, inhibits coronavirus replication at its physiologic concentration
Authors: Collins A. R. , Grubb A.
Published: March 2008

This study was conducted to examine the effect of cystatin D, a newly discovered salivary cysteine protease inhibitor, on human coronavirus replication. When MRC‐5, human diploid lung cells, were incubated with dilutions of recombinant human cystatin D from 0.65–10 μM for 1 h prior to, during and after infection with coronavirus OC43 and 229e strains, a decrease in virus yield was observed resulting in an IC50 of 0.8 μM for both virus strains. This dose is within the normal concentration range of cystatin D, 0.12–1.9 μM found in saliva. When a single dose, 2.5 μM, was applied, cystatin inhibition of release of virus progeny was not overcome until three days post infection whereas inhibition by leupeptin, a serine and cysteine protease inhibitor, was completely abrogated by two days. When cellular toxicity was measured by 3H‐thymidine uptake, cystatin D did not markedly affect cell proliferation below a 10 μM dose. The results demonstrate that cystatin D is a potent inhibitor of coronavirus replication.
Exogenous ammonium inhibits petal pigmentation and expansion in Gerbera hybrida
Authors: Huang Zhigang , Liang Minting , Peng Jianzong , Xing Tim , Wang Xiaojing
Published: March 2008

Petal pigmentation is the most important aspect in natural flower coloration. In the present study, the inhibition of petal pigmentation by exogenous ammonium was investigated. Ray floret petals detached from inflorescences of Gerbera hybrida (Shenzhen No. 5) were cultured in vitro on media supplied with different forms of nitrogen and its assimilated compounds. The expression of a set of genes involved in anthocyanin biosynthesis and regulation was determined by Northern blotting assay. It was found that ammonium (), not nitrate (), in millimolar concentrations inhibited anthocyanin accumulation. The expressions of Gerbera chalcone synthase 1 (GCHS1), Gerbera chalcone synthase 2 (GCHS2) and Gerbera dihydroflavonol‐4‐reductase (GDFR) decreased, while six other related genes showed no significant changes after treatment. Further studies on function indicated that glutamine (Gln) acted as a downstream factor of to suppress petal pigmentation. Both exogenous Gln and were found to inhibit anthocyanin accumulation in the petals, and the application of Gln was also found to inhibit the expressions of GCHS1, GCHS2 and GDFR. The application of also resulted in an increase in the activity of Gerbera glutamine synthetase (EC 6.3.1.2) along with a rapid increase of Gln content. When methionine sulfoximine, an inhibitor of glutamine synthetase (GS), was added, it was found to block the ‐induced inhibition of pigmentation. From these experiments, we conclude that the ‐induced suppression of petal pigmentation is not because of toxicity, and the inhibition of pigmentation caused by the addition of exogenous is the result of its assimilation into Gln.
Non‐photochemical loss in PSII in high‐ and low‐light‐grown leaves of Vicia faba quantified by several fluorescence parameters including LNP, , a novel parameter
Authors: Stefanov Detelin , Terashima Ichiro
Published: March 2008

Using the expression of fluorescence originated from the PSII open reaction center in the light by Oxborough and Baker (1997), we obtained a formula that expresses relationships between the quantum efficiency of PSII photochemistry in the dark (Φm= Fv/Fm) and in the light , where denotes the quantum yield of light induced non‐photochemical losses (heat dissipation and fluorescence de‐excitation) in PSII. Using LNP and other conventional fluorescence parameters, we conducted quenching analyses with leaves of broad bean plants (Vicia faba L.) grown at 700 (high light; HL) and 80 μmol photons m−2 s−1 (low light; LL). We also examined whether behavior of q0 quenching is related to the reaction center quenching. When the actinic light (AL) was strong, Stern‐Volmer quenching and LNP increased rapidly and then decreased slowly in HL leaves, while, in LL leaves, they increased slowly. It is probable that rapid formation of a large proton gradient was responsible for sharp rises in both parameters in HL leaves. The steady‐state ‘excess’ parameter [ΦEx= (1 – qP) Φm/(Φm+ LNP)], fraction of energy migrating to closed PSII centers, increased with the photon flux density of AL in LL leaves. In contrast, in HL leaves, ΦEx did not increase markedly. The examination of the relationship between ΦEx and LNP obtained at various AL revealed that in LL leaves the increase in (1 – qP) with the increase in AL prevailed, while, in HL leaves, the increase in LNP suppressed the increase in (1 – qP). Using the difference between LNP and LD (ΦND= LNP– LD, where LD= F0/Fm), q0 and qN () were calculated without using measured . The relationships between q0 and qN thus obtained for various AL levels were almost identical for both HL and LL leaves, implying no difference in the fluorescence origin between the HL and LL leaves. Usefulness of these equations expressing non‐photochemical loss is discussed.
Efficacy and safety of intravenous iron therapy as an alternative/adjunct to allogeneic blood transfusion
Authors: Muñoz M. , Breymann C. , García‐Erce J. A. , Gómez‐Ramírez S. , Comin J. , Bisbe E.
Published: March 2008

Anaemia is a common condition among patients admitted to hospital medicosurgical departments, as well as in critically ill patients. Anaemia is more frequently due to absolute iron deficiency (e.g. chronic blood loss) or functional iron deficiency (e.g. chronic inflammatory states), with other causes being less frequent. In addition, preoperative anaemia is one of the major predictive factors for perioperative blood transfusion. In surgical patients, postoperative anaemia is mainly caused by perioperative blood loss, and it might be aggravated by inflammation‐induced inhibition of erythropoietin and functional iron deficiency (a condition that cannot be corrected by the administration of oral iron). All these mechanisms may be involved in the anaemia of the critically ill. Intravenous iron administration seems to be safe, as very few severe side‐effects were observed, and may result in hastened recovery from anaemia and lower transfusion requirements. However, it is noteworthy that many of the recommendations given for intravenous iron treatment are not supported by a high level of evidence and this must be borne in mind when making decisions regarding its application to a particular patient. Nonetheless, this also indicates the need for further large, randomized controlled trials on the safety and efficacy of intravenous iron for the treatment of anaemia in different clinical settings.
Platelet‐rich plasma and platelet gel preparation using Plateltex®
Authors: Mazzucco L. , Balbo V. , Cattana E. , Borzini P.
Published: March 2008

Background  The platelet gel is made by embedding concentrate platelets within a semisolid (gel) network of polymerized fibrin. It is believed that this blood component will be used more and more in the treatment of several clinical conditions and as an adjunctive material in tissue engineering. Several systems are available to produce platelet‐rich plasma (PRP) for topical therapy. Recently, a new system became commercially available, Plateltex®. Here we report the technical performance of this system in comparison with the performance of other commercially available systems: PRGF®, PRP‐Landesber, Curasan®, PCCS®, Harvest®, Vivostat®, Regen® and Fibrinet®.

Material and Methods  Both the PRP and the gel were prepared according to the manufacturer's directions. The blood samples of 20 donors were used. The yield, the efficiency, and the amount of platelet‐derived growth factor AB (PDGF‐AB), transforming growth factor β, vascular endothelial growth factor and fibroblast growth factor were measured in the resulting PRP. The feature of the batroxobin‐induced gelation was evaluated.

Results  The yield, the collection efficiency and the growth factor content of Plateltex® were comparable to those of most of the other available systems. The gelation time was not dependent on the fibrinogen concentration; however, it was strongly influenced by the contact surface area of the container where the clotting reaction took place (P < 0·0001).

Conclusions  Plateltex® provided platelet recovery, collection efficiency and PDGF‐AB availability close to those provided by other systems marketed with the same intended use. Batroxobin, the enzyme provided to induce gelation, acts differently from thrombin, which is used by most other systems. Platelets treated with thrombin become activated; they release their growth factors quickly. Furthermore, thrombin–platelet interaction is a physiological mechanism that hastens the clot‐retraction rate. On the contrary, platelets treated with batroxobin do not become activated; they are passively entrapped within the fibrin network, and their growth factor release occurs slowly. In these conditions, the clot retraction takes longer to occur. According to these differences between thrombin and batroxobin, it is expected that batroxobin‐induced PRP activation will tailor slow release of the platelet content, thus, providing longer in loco availability of trophic factors. In selected clinical conditions, this durable anabolic factor availability might be preferable to quick thrombin‐induced growth factor release.
Cholesterol and sphingolipids in alcohol‐induced liver injury
Authors: Fernández Anna , Colell Anna , Garcia‐Ruiz Carmen , Fernandez‐Checa José C
Published: March 2008

The pathogenesis of alcohol‐induced liver disease (ALD) is still poorly understood. One of the clues to its progression relates to the alcohol‐mediated susceptibility of hepatocytes to cell death by reactive oxygen species (ROS) and inflammatory cytokines. Tumor necrosis factor α (TNF) has been considered a key ALD mediator with acidic sphingomyelinase (ASMase)‐mediated ceramide generation playing a critical role. TNF receptor 1 and 2 knock‐out mice or ASMase−/− mice exhibit resistance to alcohol‐mediated fatty liver and cell death. Furthermore, alcohol feeding has been shown to sensitize hepatocytes to TNF due to the limitation of mitochondrial glutathione (mGSH) through impaired import of GSH from the cytosol due to altered membrane order parameter caused by mitochondrial cholesterol increase. Selective pharmacological depletion of mGSH sensitizes hepatocytes to TNF‐mediated cell death, which reproduces the observations found with alcohol feeding. TNF signaling analyses in hepatocytes with or without mGSH depletion indicate that mGSH prevents cardiolipin peroxidation (CLOOH) formation by TNF‐induced ROS via ASMase and that CLOOH cooperates with oligomerized Bax to cause mitochondrial outer membrane permeabilization through destabilization of the lipid bilayer via increased bilayer‐to‐inverted hexagonal phase transitions. Thus, activation of ASMase and cholesterol‐mediated mGSH depletion both collaborate to promote alcohol‐induced TNF‐mediated hepatocellular damage, suggesting novel therapeutic opportunities in ALD.
Protein kinase C in the pancreatic acinar cell
Authors: Gorelick Fred , Pandol Stephen , Thrower Edwin
Published: March 2008

Pathological activation of selective signaling molecules within the pancreatic acinar cell mediates the development of acute pancreatitis. Some of the key early acinar cell events include activation of proteases, inhibition of apical secretion, and elaboration of inflammatory mediators. Previous studies have shown that supraphysiological concentrations of cholecystokinin (CCK) that can cause pancreatitis in vivo, also initiate these pathological responses in dispersed groups of acinar cells (acini). Protein kinase C (PKC) regulates many cellular events and a role for this family of signaling molecules has been described in some of the pathological responses of pancreatitis. Notably, ethanol can activate specific PKC isoforms and sensitize the acinar cells to the pathological effects of CCK. Our preliminary studies in isolated pancreatic acini and a cell‐free reconstitution system suggest that PKC can mediate protease activation in the acinar cell. These findings may be relevant to the pathogenesis of pancreatitis from alcohol and other etiologies.
S‐Adenosylmethionine in cell growth, apoptosis and liver cancer
Authors: Lu Shelly C , Mato José M
Published: March 2008

S‐Adenosylmethionine (SAMe), the principal biological methyl donor, is synthesized from methionine and ATP in a reaction catalyzed by methionine adenosyltransferase (MAT). In mammals, two genes (MAT1A and MAT2A), encode for two homologous MAT catalytic subunits, while a third gene MAT2β, encodes for the β‐subunit that regulates MAT2A‐encoded isoenzyme. Normal liver expresses MAT1A, whereas extrahepatic tissues express MAT2A. MAT2A and MAT2β are induced in human hepatocellular carcinoma (HCC), which facilitate cancer cell growth. Patients with cirrhosis of various etiologies, including alcohol, have decreased hepatic MAT activity and SAMe biosynthesis. Consequences of hepatic SAMe deficiency as illustrated by the Mat1a knock‐out mouse model include increased susceptibility to steatosis and oxidative liver injury, spontaneous development of steatohepatitis and HCC. Predisposition to HCC can be partly explained by the effect of SAMe on growth. Thus, SAMe inhibits the mitogenic effect of growth factors such as hepatocyte growth factor and, following partial hepatectomy, a fall in SAMe level is required for the liver to regenerate. During liver regeneration, the fall in hepatic SAMe is transient. If the fall were to persist, it would favor a proliferative phenotype and, ultimately, development of HCC. Not only does SAMe control liver growth, it also regulates apoptosis. Interestingly, SAMe is anti‐apoptotic in normal hepatocytes but pro‐apoptotic in liver cancer cells. In liver cancer cells but not in normal human hepatocytes, SAMe can selectively induce Bcl‐xS, an alternatively spliced isoform of Bcl‐xL that promotes apoptosis. This should make SAMe an attractive agent for both chemoprevention and treatment of HCC.
Epigenetic regulation of hepatic stellate cell activation
Authors: Mann Derek A , Mann Jelena
Published: March 2008

Activation of hepatic stellate cells (HSC) to a myofibroblast‐like phenotype is the pivotal event in liver fibrosis. In uninjured liver, HSC are quiescent and non‐dividing, but upon liver injury these cells undergo a dramatic change in phenotype which generates activated myofibroblast‐like HSC. The change in phenotype is underpinned by a global change in gene expression with hundreds of genes being up‐ or downregulated. Molecular events that orchestrate changes in gene expression take place at the level of chromatin packaging which is altered through three main processes: histone modifications, DNA methylation, and silencing by non‐coding RNAs. The present review focuses on the epigenetic regulation of HSC activation.
Transforming growth factor‐β and hepatocyte transdifferentiation in liver fibrogenesis
Authors: Meindl‐Beinker Nadja M , Dooley Steven
Published: March 2008

Currently, hepatic stellate cells (HSC) are thought to be the major fibrotic precursor cells that transdifferentiate to fibrogenic, extracellular matrix producing myofibroblasts in inflammatory liver tissue upon transforming growth factor‐β (TGF‐β) signaling, whereas hepatocytes are thought to respond with apoptosis to this cytokine. Starting out from in vitro experiments with primary hepatocyte cultures and immortalized AML‐12 cells, TGF‐β signaling in this cell type was assessed and apoptosis was found to be only a minor effect. Instead, hepatocytes undergo epithelial mesenchymal transition (EMT), a physiological process in embryogenesis and of relevance for cancerous cell transformation. In injured liver, however, this process contributes to the promotion of fibrosis. Already after a few days of culture, hepatocytes lose their epithelial honeycomb‐like shape towards a fibroblast‐like phenotype. We could demonstrate by microarray analysis that stimulation of hepatocytes with TGF‐β regulates the expression of genes involved in EMT and fibrosis. Among these were, for example, Snail, a known mediator of EMT, and connective tissue growth factor (CTGF), a strong inducer of fibrosis. In a mouse model, hepatocyte‐specific overexpression of Smad7 was able to blunt a fibrogenic response after CCl4 intoxication. These results emphasize the dynamic nature of liver fibrosis, challenge the paradigm of HSC as a crucial source of liver myofibroblasts and hint towards a prominent role for hepatocytes in liver fibrogenesis.
Quantifying the risk for alcohol‐use and alcohol‐attributable health disorders: Present findings and future research needs
Authors: Li Ting‐Kai
Published: March 2008

The aim of the present review was to: (i) highlight epidemiological and other studies that have generated important data on the harmful patterns of drinking that increase the risk for chronic diseases, including alcohol dependence, and on the mechanisms by which alcohol produces and, in some instances, may protect against damage; and (ii) discuss a conceptual basis for quantifying risk criteria for alcohol‐induced chronic disease based on the quantity, frequency, and pattern of drinking. The relationship between heavy drinking and risk for adverse health conditions such as alcoholic liver disease (ALD), dementia, and alcohol dependence is well known. However, not everyone who drinks chronically develops ALD or dementia, and the major risk factors for disease development and the mechanisms by which this occurs have remained unclear. Large‐scale, general population‐based studies have provided the evidence by which quantifying the frequency of a pattern of high‐risk drinking can be related directly to risk and the severity of alcohol dependence. Cellular and molecular biology studies have identified the major pathways of alcohol metabolism and how genetics and the environment can interact in some individuals to further increase the risk of organ damage. Extant databases should allow scientists and clinicians jointly to develop the framework for quantifying the drinking patterns that increase the risk of alcohol‐induced organ pathologies, to develop clinical practice guidelines, such as those used to diagnose other common complex diseases (e.g. diabetes and hypertension), and to propose future studies for refining such guidelines. Attention must be paid to comorbid conditions such as hepatitis B and C infections, HIV, obesity, and environmental exposures other than alcohol. Developing trait and state biomarkers is critical to the process of discovery and to fulfilling the promise of personalized medicine.