bluuebunny/crossref_metadata_2025 huggingface dataset:
Gastric cancer screening of a high‐risk population in Japan using serum pepsinogen and barium digital radiography
Authors: Ohata Hiroshi , Oka Masashi , Yanaoka Kimihiko , Shimizu Yasuhito , Mukoubayashi Chizu , Mugitani Kouichi , Iwane Masataka , Nakamura Hideya , Tamai Hideyuki , Arii Kenji , Nakata Hiroya , Yoshimura Noriko , Takeshita Tetsuya , Miki Kazumasa , Mohara Osamu , Ichinose Masao
Published: September 2005

With the aim of developing more efficient gastric cancer screening programs for use in Japan, we studied a new screening program that combines serum pepsinogen (PG) testing and barium digital radiography (DR). A total of 17 647 middle‐aged male subjects underwent workplace screening over a 7‐year period using a combination of PG testing and DR. This program's effectiveness, as well as other characteristics of the program, was analyzed. Forty‐nine cases of gastric cancer were detected (comprising 88% early cancer cases). The detection rate was 0.28%, and the positive predictive value was 0.85%. The PG test detected 63.3% of cases, DR detected 69.4% of cases, and both tests were positive in 32.7% of cancer cases. The two methods were almost equally effective, and were considerably more effective than conventional screening using photofluorography. Each screening method detected a distinct gastric cancer subgroup; the PG test efficiently detected asymptomatic small early cancer with intestinal type histology, while DR was efficient at detecting cancers with depressed or ulcerated morphology and diffuse type histology. The cost for the detection of a single cancer was much less than that for conventional screening. In fact, it is possible to further reduce the cost of detecting a single cancer to a cost comparable to that of surgically resecting a single gastric cancer. Thus, it is probable that a highly efficient gastric cancer screening system can be implemented by combining the two screening methods. Such a screening program would be beneficial in a population at high risk for gastric cancer. (Cancer Sci 2005; 96: 713 – 720)
Acupressure for prevention of pre‐operative anxiety: a prospective, randomised, placebo controlled study
Authors: Agarwal A. , Ranjan R. , Dhiraaj S. , Lakra A. , Kumar M. , Singh U.
Published: September 2005

Pre‐operative anxiety is associated with many unwanted effects such as increased analgesic and anaesthetic requirement, postoperative pain and prolonged hospital stay. In the present study, we investigated the effects of acupressure on pre‐operative anxiety and bispectral index (BIS) values. Seventy‐six adults, ASA grade I and II, undergoing elective surgery, were randomly assigned to two equal groups. Group 1 (control) received acupressure at an inappropriate site and group 2 (acupressure) received acupressure at extra 1 point. The study was conducted during the pre‐operative period and the duration of the study was 40 min (acupressure was applied for 10 min and thereafter patients were observed for another 30 min). Anxiety was recorded on a visual stress scale (VSS) at the start of the study and thereafter at 10 and 40 min. BIS was recorded at 0, 2, 5, 10, 12, 15, 30 and 40 min. The VSS decreased in both groups following pressure application for 10 min: median VSS (interquartile range) were 5 (1) vs. 8 (1) in the acupressure and 7 (0) vs. 8 (1) in the control groups (p < 0.001). Both pre‐operative anxiety and BIS decreased significantly during acupressure application at extra 1 point (p < 0.001). Acupressure is effective in decreasing both pre‐operative anxiety and BIS; however, these effects are not sustained 30 min following release of acupressure. Further studies are needed to elucidate the duration for which acupressure is effective.
Influence of Mediterranean sea‐level changes on the Dacic Basin (Eastern Paratethys) during the late Neogene: the Mediterranean Lago Mare facies deciphered
Authors: Clauzon Georges , Suc Jean‐Pierre , Popescu Speranta‐Maria , Marunteanu Mariana , Rubino Jean‐Loup , Marinescu Florian , Melinte Mihaela Carmen
Published: August 2005

A recently published scenario viewing the Messinian salinity crisis as two evaporitic steps rather than one has led to a search for new indices of the crisis in the Eastern Paratethys. Fluvial processes characterized the southwestern Dacic Basin (Southern Romania, i.e. the Carpathian foredeep) whereas brackish sediments were continuously deposited in its northern part. This is consistent with previously evidenced responses of the Black Sea to the Messinian salinity crisis. High sea‐level exchanges between the Mediterranean Sea and Eastern Paratethys are considered to have occurred just before and just after desiccation of the Mediterranean. This accounts for two successive Mediterranean nannoplankton‐dinocyst influxes into the Eastern Paratethys that, respectively, belong to zones NN 11 and NN 12. Meanwhile, two separate events that gave rise to Lago Mare facies (with Paratethyan Congeria, ostracods and/or dinoflagellate cysts) arose in the Mediterranean Basin in response to these high sea‐level exchanges and located 5.52 and 5.33 Ma (isotopic stages TG 11 and TG 5, respectively), i.e. just before and just after the almost complete desiccation of the Mediterranean). These Lago Mare facies formed independently of lakes with ostracods of the Cyprideis group that developed in the central basins during the final stages of desiccation. The gateway faciliting these water exchanges is not completely identified. A proto‐Bosphorus strait seems unlikely. A plausible alternative route extends from the northern part of the Thessaloniki region up to the Dacic Basin and through Macedonia and the Sofia Basin. The expression ‘Lago Mare’ is chronostratigraphically ambiguous and should be discontinued for this purpose, although it might remain useful as a palaeoenvironmental term.
Migration pathways in the Central North Slope foreland basin, Alaska USA: solute and thermal constraints on fluid flow simulations
Authors: Nunn Jeffrey A. , Hanor Jeffrey S. , Lee Youngmin
Published: August 2005

The North Slope foreland basin, Alaska, USA is an east–west asymmetrical trough‐shaped basin adjacent to the Brooks Range fold‐thrust mountain belt. Lower Cretaceous age rocks make up much of the sediment fill, including flysch‐like marine turbidites and shales of the Torok and Fortress Mountain formations and marine and sandstones, shales and conglomerates of the overlying Nanushuk group. Lower Cretaceous age rocks were deposited on top of a Palaeozoic and Mesozoic age passive margin sequence. We have conducted numerical simulations of fluid flow driven by topographic recharge in the Central North Slope foreland basin. These simulations are constrained by salinity estimates from well logs, location of oil and gas fields, vitrinite reflectance and heat flow measurements. Our model results indicate that there are two south to north pathways for fluid migration. The primary pathway for fluid movement is downward through the Fortress Mountain formation, then upwards along the interface between the Fortress Mountain and Torok Formation and finally northward through the permeable Nanushuk group. A smaller mass of groundwater moves along sands below the Torok formation and into offshore sediments north of Alaska. Very little meteoric water enters the underlying Palaeozoic rocks in our simulations, which could explain the presence of deep saline pore waters. Our results also show that permafrost is a primary control on the pathway and rate of fluid flow by controlling the distribution of surface recharge and discharge. For example, areas of high heat flow and low saline waters along the arctic coast may represent upward groundwater discharge because of the absence of permafrost. As surface temperatures were warmer in the Miocene, the absence of permafrost would produce a more local fluid circulation pattern and less transfer of heat energy from south to north.
Dynamic modelling of passive margin salt tectonics: effects of water loading, sediment properties and sedimentation patterns
Authors: Gemmer Lykke , Beaumont Christopher , Ings Steven J.
Published: August 2005

We investigate the evolution of passive continental margin sedimentary basins that contain salt through two‐dimensional (2D) analytical failure analysis and plane‐strain finite‐element modelling. We expand an earlier analytical failure analysis of a sedimentary basin/salt system at a passive continental margin to include the effects of submarine water loading and pore fluid pressure. Seaward thinning sediments above a weak salt layer produce a pressure gradient that induces Poiseuille flow in the viscous salt. We determine the circumstances under which failure at the head and toe of the frictional–plastic sediment wedge occurs, resulting in translation of the wedge, landward extension and seaward contraction, accompanied by Couette flow in the underlying salt. The effects of water: (i) increase solid and fluid pressures in the sediments; (ii) reduce the head to toe differential pressure in the salt and (iii) act as a buttress to oppose failure and translation of the sediment wedge. The magnitude of the translation velocity upon failure is reduced by the effects of water.

The subsequent deformation is investigated using a 2D finite‐element model that includes the effects of the submarine setting and hydrostatic pore pressures. The model quantitatively simulates a 2D approximation of the evolution of natural sedimentary basins on continental margins that are formed above salt. Sediment progradation above a viscous salt layer results in formation of landward extensional basins and listric normal growth faults as well as seaward contraction. At a later stage, an allochthonous salt nappe overthrusts the autochthonous limit of the salt. The nature and distribution of major structures depends on the sediment properties and the sedimentation pattern. Strain weakening of sediment favours landward listric growth faults with formation of asymmetric extensional depocentres. Episodes of low sediment influx, with partial infill of depocentres, produce local pressure gradients in the salt that result in diapirism. Diapirs grow passively during sediment aggradation.
Co‐stimulatory pathways in lymphocyte regulation: the immunoglobulin superfamily
Authors: Peggs Karl S. , Allison James P.
Published: August 2005

The controlled orchestration of immune responses is a vital feature of cellular immunity in a system that must be able to reliably distinguish self from non‐self. Contrary to early beliefs, peptide recognition by T cells exhibits a relatively high level of promiscuity. The requirement for a second signalling event to be present in addition to that provided by T cell receptor ligation for T cell activation to proceed helps to prevent inappropriately directed responses. An expanding array of co‐stimulatory or inhibitory signalling receptors and ligands are now recognised to be involved in the control of the crucial decisions made determining the activation, expansion, and effector functions of responding cells, and ultimately the final targeting and execution of these functions. Tight regulation of the temporal and spatial organisation of receptor/ligand expression, combined with both forward and reverse signalling, endows an extraordinary elegance to these co‐stimulatory pathways. The immunoglobulin superfamily occupies a central importance in this coordination of immune responses. The understanding of its relevance in a variety of physio‐pathological circumstances is now yielding a number of potential targets for therapeutic manipulation, and such immunological molecular adjuvants are beginning to enter clinical trials.
Autologous stem cell transplantation in the treatment of adults with acute myeloid leukaemia
Authors: Breems Dimitri A. , Löwenberg Bob
Published: August 2005

Most adult patients under 60 years with acute myeloid leukaemia (AML) who achieve a complete remission after induction chemotherapy will relapse if they do not receive further therapy. Consolidation treatment with autologous stem cell transplantation (SCT) is one option that has been studied extensively. High‐dose cytotoxic therapy followed by autologous SCT or intensive cycles of chemotherapy furnish overall approximately similar probabilities of survival when applied in first remission. Here, we present a concise update regarding the place of autologous SCT in the treatment of AML. Particular issues discussed are the value of autologous SCT in different prognostic subsets of AML and the value of autologous mobilised peripheral blood stem cell transplants, which offer a much faster haematopoietic recovery.
Sir John Dacie MD, FRS, FRCP, FRCPath
Authors: Lewis Mitchell
Published: August 2005

Prof. Sir John Dacie was one of the most distinguished haematologists of the 20th century. He died on 12 February 2005 at the age of 92. This annotation is intended to give an impression of his career, and his role in the development of haematology in the UK and beyond. It describes his approach to haematological practise, taking account of both clinical and laboratory aspects, and reviews his published works over a range of haematological topics.
Selective apoptosis of natural killer‐cell tumours by l‐asparaginase
Authors: Ando Miki , Sugimoto Koichi , Kitoh Toshiyuki , Sasaki Makoto , Mukai Kouichi , Ando Jun , Egashira Motoki , Schuster Sheldon M. , Oshimi Kazuo
Published: August 2005

We examined the effectiveness of various anti‐tumour agents to natural killer (NK)‐cell tumour cell lines and samples, which are generally resistant to chemotherapy, using flow cytometric terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end‐labelling (TUNEL) assay. Although NK‐YS and NK‐92 were highly resistant to various anti‐tumour agents, l‐asparaginase induced apoptosis in these two NK‐cell lines. NK‐cell leukaemia/lymphoma and acute lymphoblastic leukaemia (ALL) samples were selectively sensitive to l‐asparaginase and to doxorubicin (DXR) respectively. Samples of chronic NK lymphocytosis, an NK‐cell disorder with an indolent clinical course, were resistant to both drugs. Our study clearly separated two major categories of NK‐cell disorders and ALL according to the sensitivity to DXR and l‐asparaginase. We examined asparagine synthetase levels by real‐time quantitative polymerase chain reaction (RQ‐PCR) and immunostaining in these samples. At least in nasal‐type NK‐cell lymphoma, there was a good correlation among asparagine synthetase expression, in vitro sensitivity and clinical response to l‐asparaginase. In aggressive NK‐cell leukaemia, although asparagine synthetase expression was high at both mRNA and protein levels, l‐asparaginase induced considerable apoptosis. Furthermore, samples of each disease entity occupied a distinct area in two‐dimensional plotting with asparagine synthetase mRNA level (RQ‐PCR) and in vitrol‐asparaginase sensitivity (TUNEL assay). We confirmed rather specific anti‐tumour activity of l‐asparaginase against NK‐cell tumours in vitro, which provides an experimental background to the clinical use of l‐asparaginase for NK‐cell tumours.
Accelerated lymphangiogenesis in malignant lymphoma: possible role of VEGF‐A and VEGF‐C
Authors: Kadowaki Ikuko , Ichinohasama Ryo , Harigae Hideo , Ishizawa Kenichi , Okitsu Yoko , Kameoka Junichi , Sasaki Takeshi
Published: August 2005

There is little information regarding the lymphangiogenesis of malignant lymphoma. In this study, we evaluated the lymphangiogenesis and angiogenesis in 44 lymph nodes of 39 malignant lymphomas and five non‐reactive normal lymph nodes, based on the lymphatic vessel density (LVD) and microvessel density (MVD) calculated by the computer‐assisted assessment of vessel density. The LVD of malignant lymphomas was significantly higher than that of non‐reactive normal lymph nodes, irrespective of subtypes (P = 0·00077). On the contrary, there was no difference in MVD between malignant lymphomas and non‐reactive normal lymph nodes, except for diffuse large B cell lymphomas, which had a significantly low value of MVD, in comparison with non‐reactive normal lymph nodes (P = 0·009). We further examined the expression of vascular endothelial growth factor (VEGF)‐C and VEGF‐A, which function on lymphangiogenesis in lymph node samples. VEGF‐C was expressed in 36 of 39 malignant lymphomas. All 39 of the malignant lymphoma samples expressed VEGF‐A. Furthermore, the level of LVD and VEGF‐A or VEGF‐C was positively correlated. These findings suggest that lymphangiogenesis is actively developed in lymph nodes of malignant lymphomas and it may be induced by both VEGF‐A and VEGF‐C secreted from lymphoma cells.
Levels of soluble endothelium‐derived adhesion molecules in patients with sickle cell disease are associated with pulmonary hypertension, organ dysfunction, and mortality
Authors: Kato Gregory J. , Martyr Sabrina , Blackwelder William C. , Nichols James S. , Coles Wynona A. , Hunter Lori A. , Brennan Marie‐Luise , Hazen Stanley L. , Gladwin Mark T.
Published: August 2005

Endothelial cell adhesion molecules orchestrate the recruitment and binding of inflammatory cells to vascular endothelium. With endothelial dysfunction and vascular injury, the levels of endothelial bound and soluble adhesion molecules increase. Such expression is modulated by nitric oxide (NO), and in patients with sickle cell disease (SCD), these levels are inversely associated with measures of NO bioavailability. To further evaluate the role of endothelial dysfunction in a population study of SCD, we have measured the levels of soluble endothelium‐derived adhesion molecules in the plasma specimens of 160 adult patients with SCD during steady state. Consistent with a link between endothelial dysfunction and end‐organ disease, we found that higher levels of soluble vascular cell adhesion molecule‐1 (sVCAM‐1) were associated with markers indicating renal dysfunction and hepatic impairment. Analysis of soluble intercellular cell adhesion molecule‐1 (sICAM‐1), sE‐selectin and sP‐selectin levels indicated partially overlapping associations with sVCAM‐1, with an additional association with inflammatory stress and triglyceride levels. Importantly, increased soluble adhesion molecule expression correlated with severity of pulmonary hypertension, a clinical manifestation of endothelial dysfunction. Soluble VCAM‐1, ICAM‐1, and E‐selectin were independently associated with the risk of mortality in this cohort. Our data are consistent with steady state levels of soluble adhesion molecules as markers of pulmonary hypertension and risk of death.
Isotype‐specific detection of ABO blood group antibodies using a novel flow cytometric method
Authors: Stussi G. , Huggel K. , Lutz H. U. , Schanz U. , Rieben R. , Seebach J. D.
Published: August 2005

Several methods to detect anti‐A/B antibodies based on haemagglutination and haemolysis have been described. These methods measure predominantly anti‐A/B immunoglobulin (Ig)M, whereas anti‐A/B IgG and IgG subclasses are less well examined. We established a flow cytometry method (ABO‐fluorescence‐activated cell sorting; ABO‐FACS) to quantify binding of anti‐A/B IgM, IgG and IgG subclasses to human A or B red blood cells. Anti‐A/B IgM were present in the majority of 120 blood donors, as expected from blood group typing. The sensitivity and specificity of anti‐A/B IgM to predict the blood group was 93% and 96% respectively. Anti‐A/B IgG was found in 34/38 blood group O samples (89%). Anti‐B IgG in blood group A or anti‐A IgG in blood group B was present in 4/28 (14%) and 1/28 (4%) samples, respectively, and absent in 26 AB sera. IgG2 was the predominant IgG subclass. The correlation of anti‐A/B IgM and IgG in the ABO‐FACS with haemagglutination titres was 0·870 and 0·783, respectively (n = 240; P < 0·001) whereas the comparison of ABO‐FACS with ABO‐enzyme‐linked immunosorbent assay was less significant. In conclusion, ABO‐FACS is a valid method to quantify anti‐A/B IgM, IgG and IgG subclasses. It opens the possibility of isotype‐specific monitoring of anti‐A/B antibodies levels after ABO‐incompatible solid organ and stem cell transplantation.
Functional osteoclast‐like transformation of cultured human myeloma cell lines
Authors: Calvani Nicola , Cafforio Paola , Silvestris Franco , Dammacco Franco
Published: August 2005

Hyperactive osteoclastogenesis is a hallmark of multiple myeloma, a B cell neoplasia homing to bone marrow and resulting in multiple osteolytic lesions and skeleton devastation. We provide evidence that myeloma cells can themselves act as osteoclastsin vitro. By extending standard cultures of U‐266 and MCC‐2 myeloma cell lines, we found that subsets of adherent cells also expressed the osteoclast phenotype, including multinuclear morphology, cytoplasmic tartrate‐resistant acid phosphatase, the calcitonin receptor and a specific osteoclast antigen. These subsets resorbed bone substrates by producing osteoclast enzymes as well as the characteristic redistribution of F‐actin in their cytoskeleton, thus forming the sealing zone that is adopted by adherent osteoclasts to generate the acidified environment essential for bone resorption. Neither the phenotype nor the functional properties of osteoclasts were detected in parental non‐adherent cells. In adherent cultures osteoclastogenesis was associated with deregulated expression of both receptor activator of nuclear transcription factor (NF)‐κB (RANK) and its ligand RANK‐L, which triggers cell maturation in osteoclast precursors. Resorption of bone substrates was prevented by a neutralising anti‐RANK‐L antibody. Our data indicate that osteoclast‐like transformation of both U‐266 and MCC‐2 cellular models of human myeloma is dependent on RANK‐L stimulation.
Nephrotic syndrome: an under‐recognised immune‐mediated complication of non‐myeloablative allogeneic haematopoietic cell transplantation
Authors: Srinivasan R. , Balow J. E. , Sabnis S. , Lundqvist A. , Igarashi T. , Takahashi Y. , Austin H. , Tisdale J. , Barrett J. , Geller N. , Childs R.
Published: September 2005

Nephrotic syndrome (NS) is an extremely rare complication of myeloablative allogeneic haematopoietic cell transplantation (HCT) that usually occurs in association with chronic graft‐versus‐host disease (C‐GVHD). We observed an unexpectedly high incidence of NS in a cohort of 163 consecutive patients undergoing non‐myeloablative HCT from a related human leucocyte antigen‐compatible donor. Seven patients developed NS at a median 318 d post‐transplant (range 119–1203 d; cumulative incidence 6·1%). The median age at onset of NS was 46 years (range 33–59 years); three of the seven patients had no evidence of C‐GVHD while four had accompanying limited C‐GVHD. At diagnosis, median proteinuria was 16·5 g/24 h (range 3–24 g/24 h). Renal biopsy was performed in four cases and revealed membranous nephropathy. NS was not always associated with other symptoms of C‐GVHD, and in contrast to previous reports, usually did not improve with the re‐initiation of aggressive immunosuppression, resulting in progressive renal failure necessitating dialysis in three of seven cases. Membranous nephropathy resulting in NS is a previously unrecognised and clinically significant complication of non‐myeloablative HCT.
Transcription repression activity is associated with the type I isoform of the MMSET gene involved in t(4;14) in multiple myeloma
Authors: Todoerti Katia , Ronchetti Domenica , Agnelli Luca , Castellani Stefano , Marelli Silvia , Deliliers Giorgio Lambertenghi , Zanella Alberto , Lombardi Luigia , Neri Antonino
Published: September 2005

The WHSC1/MMSET gene, involved in t(4;14)(p16.3;q32) in multiple myeloma, encodes putative isoforms (MMSET I, MMSET II and RE‐IIBP) which are thought to be involved in transcription regulation. We investigated their activity in transfected 293T and HeLa cells. Both MMSET I and MMSET II were localised in the nucleus, whereas RE‐IIBP showed cytoplasmic and nucleolar staining. MMSET I dose‐dependently repressed the transcriptional activity of the promoter region of the thymidine kinase gene, whereas MMSET II and RE‐IIBP had no effect. The HDAC inhibitor, trichostatin A, reduced MMSET I repression activity and in vitro co‐immunoprecipitation analyses indicated that MMSET I specifically recruits HDAC1 and mSin3b, but not HDAC2 or HDAC4. Our data support the hypothesis that MMSET may act as a transcription regulator; different functions may be associated with distinct isoforms.
Pax6 is required for production and maintenance of progenitor cells in postnatal hippocampal neurogenesis
Authors: Maekawa Motoko , Takashima Noriko , Arai Yoko , Nomura Tadashi , Inokuchi Kaoru , Yuasa Shigeki , Osumi Noriko
Published: August 2005

Neurogenesis is crucial for brain formation and continues to take place in certain regions of the postnatal brain including the subgranular zone (SGZ) of the hippocampal dentate gyrus (DG). Pax6 transcription factor is a key player for patterning the brain and promoting embryonic neurogenesis, and is also expressed in the SGZ. In the DG of wild‐type rats, more than 90% of total BrdU‐incorporated cells expressed Pax6 at 30 min time point after BrdU injection. Moreover, approximately 60% of Pax6+ cells in the SGZ exhibited as GFAP+ cells with a radial glial phenotype and about 30% of Pax6+ cells exhibited as PSA‐NCAM+ cells in clusters. From BrdU labeling for 3 days, we found that cell proliferation was 30% decreased at postnatal stages in Pax6‐deficient rSey2/+ rat. BrdU pulse/chase experiments combined with marker staining revealed that PSA‐NCAM+ late progenitor cells increased at the expense of GFAP+ early progenitors in rSey2/+ rat. Furthermore, expression of Wnt ligands in the SGZ was markedly reduced in rSey2/+ rat. Taken all together, an appropriate dosage of Pax6 is essential for production and maintenance of the GFAP+ early progenitor cells in the postnatal hippocampal neurogenesis.
Rho mediates endocytosis of epidermal growth factor receptor through phosphorylation of endophilin A1 by Rho‐kinase
Authors: Kaneko Takako , Maeda Akio , Takefuji Mikito , Aoyama Hiroki , Nakayama Masanori , Kawabata Saeko , Kawano Yoji , Iwamatsu Akihiro , Amano Mutsuki , Kaibuchi Kozo
Published: August 2005

After binding of epidermal growth factor (EGF), the EGF receptor is activated, internalized by endocytosis, and subsequently degraded in the lysosomal pathway. Endocytotic trafficking of the activated EGF receptor is essential for controlling EGF signaling. Upon ligand‐induced activation of EGF receptors, Cbl (ubiquitin ligase) binds to the activated receptor and leads to translocation of the CIN85 (Cbl‐interacting protein of 85 kDa)/endophilin complex in the vicinity of the activated EGF receptors. Endophilin is known as a key regulator of clathrin‐mediated endocytosis, and the translocation of endophilin in the vicinity of active EGF receptor is thought to promote receptor internalization. The constitutively active mutant of small GTPase Rho inhibits EGF receptor endocytosis. In this study, we found that this inhibitory effect was canceled by the dominant negative form of Rho‐associated kinase (Rho‐kinase), which is an effector of Rho. To clarify the molecular mechanisms of endocytosis downstream of Rho/Rho‐kinase signal, we searched for and identified endophilin A1 as a novel substrate of Rho‐kinase. We identified the phosphorylation site of endophilin A1 at Thr‐14 and made endophilin T14D (substitution of Thr‐14 by Asp), which is expected to mimic the phosphorylation state of endophilin A1. Endophilin T14D inhibited EGF receptor internalization. Furthermore, phosphorylation of endophilin by Rho‐kinase inhibited the binding to CIN85. Taken together, these results suggest that Rho‐kinase phosphorylates endophilin downstream of Rho and regulates EGF receptor endocytosis through the inhibition of binding between endophilin and CIN85.
Specific N‐terminal interactions of the Escherichia coli SeqA protein are required to form multimers that restrain negative supercoils and form foci
Authors: Odsbu Ingvild , Klungsøyr Hege K. , Fossum Solveig , Skarstad Kirsten
Published: September 2005

The Escherichia coli SeqA protein binds preferentially to hemimethylated DNA and is required for inactivation (sequestration) of newly formed origins. A mutant SeqA protein, SeqA4 (A25T), which is deficient in origin sequestration in vivo, was found here to have lost the ability to form multimers, but could bind as dimers with wild‐type affinity to a pair of hemimethylated GATC sites. In vitro, binding of SeqA dimers to a plasmid first generates a topology change equivalent to a few positive supercoils, then the binding leads to a topology change in the “opposite” direction, resulting in a restraint of negative supercoils. Binding of SeqA4 mutant dimers produced the former effect, but not the latter, showing that a topology change equivalent to positive supercoiling is caused by the binding of single dimers, whereas restraint of negative supercoils requires multimerization via the N‐terminus. In vivo, mutant SeqA4 protein was not capable of forming foci observed by immunofluorescence microscopy, showing that N‐terminus–dependent multimerization is required for building SeqA foci. Overproduction of SeqA4 led to partially restored initiation synchrony, indicating that origin sequestration may not depend on efficient higher‐order multimerization into foci, but do require a high local concentration of SeqA.
Both antigen optimization and lysosomal targeting are required for enhanced anti‐tumour protective immunity in a human papillomavirus E7‐expressing animal tumour model
Authors: Suk Kim Mi , Sin Jeong‐Im
Published: August 2005

DNA immunization is a new approach for cancer immune therapy. In this study, we constructed human papillomavirus (HPV) 16 E7 expression vector cassettes and then compared the abilities of these constructs to induce antitumour protection. Lysosome‐targeted E7 antigens, and to a lesser degree signal sequence‐conjugated and transmembrane region sequence‐conjugated E7 antigens in a DNA form, displayed tumour protection significantly higher than wild‐type E7 antigens. This enhanced tumour protection was mediated by CD8+ cytotoxic T lymphocytes (CTL), as determined by in vivo T‐cell depletion and in vitro interferon‐γ (IFN‐γ) production. Subsequent co‐injection with interleukin‐12‐expressing cDNA showed insignificantly enhanced antitumour protection. However, E7 codon optimization plus lysosomal targeting resulted in a dramatic enhancement in antitumour protection both prophylactically and therapeutically through augmentation of the E7‐specific CTL population, compared to either one of them alone. However, wild‐type or codonoptimized E7 antigens without intracellular targeting displayed no protection against tumour challenge. Thus, these data suggest that antigen codon optimization plus lysosomal targeting strategy could be important in crafting more efficacious E7 DNA vaccines for tumour protection.
Quality retention during the chilled distribution of farmed turbot (Psetta maxima): effect of a primary slurry ice treatment
Authors: Piñeiro Carmen , Bautista Raquel , Rodríguez Óscar , Losada Vanesa , Barros‐Velázquez Jorge , Aubourg Santiago P.
Published: August 2005

The quality and shelf‐life of chilled farmed turbot (Psetta maxima) was evaluated by sensory, microbiological and biochemical procedures after being subjected to a two‐step refrigeration storage in slurry ice and flake ice, respectively. Turbot specimens were stored for 10 or 17 days in slurry ice, and then were transferred to flake ice for 1–3 days to simulate the sale conditions in the market. The results were compared with control batches stored only in flake ice and processed in parallel. Storage of turbot in the two‐step strategy resulted in a better maintenance of sensory quality, especially with regard to its mucus production and gill odour development, better control of microbial activity, especially of aerobes, and the slowing down of some biochemical degradation mechanisms such as the nucleotide degradation pathway and trimethylamine production. As a consequence, the shelf‐life was extended significantly. From these results it can be concluded that the refrigerated transport of farmed turbot in slurry ice enhances its shelf‐life, before its transfer into flake ice in the retail market.
Effect of flour fortification with haem liposome on bread and bread doughs
Authors: Albaldawi Amira , Brennan Charles S. , Alobaidy Khalid , Alammar Waffa , Aljumaily Dhamla
Published: August 2005

Slaughter blood haem was encapsulated in lecithin : cholesterol Liposomes. Wheat flour was fortified with this mixture at two levels (60 and 100 mg/100 g flour) using the haem liposomes as iron source. The effect of haem fortification on gluten and fat content of flours was determined, as were the effects of fortification on dough visco‐elastic characteristics (water absorption, development time, dough stability and breakdown time). The baking properties of the breads were also investigated. The addition of haem liposomes increased the fat content of flours, and had a positive effect on the stability and rheological characteristics of the dough. Loaf volume and crumb uniformity was improved. These results indicate a potential nutritional use for haem liposomes..
The preservation of the quality of the muscle in frozen Australian red claw crayfish (Cherax quadricarinatus) by pre‐storage anti‐oxidant dipping treatments†
Authors: Tseng Yen‐Chang , Xiong Youling L. , Webster Carl D.
Published: August 2005

Australian red claw crayfish tails were dipped in either 0.06% (w/w) anti‐oxidant solutions (tocopherols, propyl gallate, or rosemary extract) or in water and subsequently stored in a −20 °C freezer. At the end of 0, 1, 3, and 6 months of storage, the raw muscle samples were analysed for lipid oxidation, the thermal stability of proteins, cooking yield and shear force. Red claw treated with anti‐oxidants showed a lower (P < 0.05) thiobarbituric acid‐reactive substances (TBARS) production than the non‐anti‐oxidant control (dipped in water). Enthalpy of denaturation (ΔH) for the myosin head and shear values for the tails decreased (P < 0.05) in all samples after 6 months of storage, while cooking yield was unaffected by storage. Overall, red claw muscle was quite stable and the oxidation, measured as TBARS values, was small (<0.3 mg kg−1) but could be further stabilized by anti‐oxidant dipping treatments. However the anti‐oxidant dipping did not prevent texture softening in red claw muscle during frozen storage.
The effect of high pressure treatment on rheological characteristics and colour of mango pulp
Authors: Ahmed Jasim , Ramaswamy Hosahalli S. , Hiremath Nikhil
Published: August 2005

The effect of high‐pressure (HP) treatment (100–400 MPa for 15 or 30 min at 20 °C) on the rheological characteristics and colour of fresh and canned mango pulps was evaluated. Differences were observed in the rheological behaviour of fresh and canned mango pulps treated with HP. Shear stress–shear rate data of pulps were well described by the Herschel–Bulkley model. The consistency index (K) of fresh pulp increased with pressure level from 100 to 200 MPa while a steady decrease was noticed for canned pulp. For fresh pulp the flow behaviour index decreased with pressure treatment whereas an increasing trend was observed with canned pulp. Storage and loss moduli of treated fresh pulp with HP increased linearly with angular frequency up to 200 MPa for a treatment time of 30 min while a steady decreasing trend was found for processed pulp. No significant variation in colour was observed during pressure treatment.
Vascular epiphyte distribution patterns: explaining the mid‐elevation richness peak
Authors: CARDELÚS CATHERINE L. , COLWELL ROBERT K. , WATKINS JAMES E.
Published: August 2005

We examined in situ diversity and distribution of vascular epiphytes, as well as site environmental variables at six sites along a continuous elevational gradient (30–2600 m a.s.l.) of old‐growth forest in Costa Rica.

A total of 555 species of vascular epiphytes from 130 genera of 53 families were identified to species or morphospecies. The ferns were the most diverse group, with 138 species, followed by orchids (112 species). Cloud forest at 1000 m was the richest site, representing the maximum of a pronounced mid‐elevation peak in epiphyte species richness.

Spatial randomizations of recorded elevational ranges suggest that the overall elevational richness pattern of most epiphyte groups on this transect is substantially influenced by the mid‐domain effect (MDE, the mid‐elevation overlap of large‐ranged species). Among the environmental factors considered (rainfall, temperature and canopy light environment), only rainfall was significantly correlated with richness.

Different patterns of richness for vascular epiphytes and for trees indicate that mechanisms differ between life forms.

We collected 26% of the estimated epiphyte species of Costa Rica along a single mountain transect. This, together with the finding that different groups and life forms varied in the elevation at which species richness peaked, highlights the need to conserve the few remaining intact elevational gradients in Latin America.
Patterns of virulence gene expression differ between biofilm and tissue communities of Streptococcus pyogenes
Authors: Cho Kyu Hong , Caparon Michael G.
Published: August 2005

The ability of Streptococcus pyogenes to form biofilm‐like bacterial communities during infection of soft tissue has suggested that the capacity to produce biofilm may be important for pathogenesis. To examine this relationship, a panel of mutants was evaluated for their ability to form biofilm on abiotic surfaces in several assays. Several established virulence factors were crucial for biofilm formation, including the M protein, required for initial cell‐surface interactions, and the hyaluronic acid capsule, required for subsequent maturation into a three‐dimensional structure. Mutants lacking the transcription regulators Mga and CovR (CsrR) also failed to form biofilm. Comparison of transcriptional profiles revealed differential regulation of approximately 25% of the genome upon adaptation to biofilm. During infection of zebrafish, several virulence factors (notably cysteine protease and streptokinase) were regulated in a biofilm‐like manner. However, the overall profile of virulence factor expression indicated that tissue communities have a pattern of gene expression different from biofilm. Taken together, these data show that while biofilm and tissue communities have many characteristics in common, that biofilm reproduces only a subset of the myriad cues used by tissue communities for regulation of virulence.
An atypical mitogen‐activated protein kinase controls cytokinesis and flagellar motility during male gamete formation in a malaria parasite
Authors: Tewari Rita , Dorin Dominique , Moon Robert , Doerig Christian , Billker Oliver
Published: August 2005

The transmission of malaria parasites to the mosquito depends critically on the rapid initiation of sexual reproduction in response to triggers from the mosquito midgut environment. We here identify an essential function for an atypical mitogen‐activated protein kinase of the rodent malaria parasite Plasmodium berghei, Pbmap‐2, in male sexual differentiation and parasite transmission to the mosquito. A deletion mutant no longer expressing the Pbmap‐2 protein develops as wild type throughout the asexual erythrocytic phase of the life cycle. Gametocytes, the sexual transmission stages, form normally and respond in vitro to the appropriate environmental cues by rounding up and emerging from their host cells. However, microgametocytes fail to release flagellated microgametes. Female development is not affected, as judged by the ability of macrogametes to become cross‐fertilized by microgametes from a donor strain. Cellular differentiation of Pbmap‐2 KO microgametocytes is blocked at a late stage of male gamete formation, after replication and mitoses have been completed and axonemes have been assembled. These data demonstrate a function for Pbmap‐2 in initiating cytokinesis and axoneme motility, possibly downstream of a cell cycle checkpoint for the completion of replication and/or mitosis, which are extraordinarily rapid in the male gametocyte.
VSG switching in Trypanosoma brucei: antigenic variation analysed using RNAi in the absence of immune selection
Authors: Aitcheson Niall , Talbot Suzanne , Shapiro Jesse , Hughes Katie , Adkin Carl , Butt Thomas , Sheader Karen , Rudenko Gloria
Published: August 2005

Trypanosoma brucei relies on antigenic variation of its variant surface glycoprotein (VSG) coat for survival. We show that VSG switching can be efficiently studied in vitro using VSG RNAi in place of an immune system to select for switch variants. Contrary to models predicting an instant switch after inhibition of VSG synthesis, switching was not induced by VSG RNAi and occurred at a rate of 10−4 per division. We find a highly reproducible hierarchy of VSG activation, which appears to be capable of resetting, whereby more than half of the switch events over 12 experiments were to one of two VSGs. We characterized switched clones according to switch mechanism using marker genes in the active VSG expression site (ES). Transcriptional switches between ESs were the preferred switching mechanism, whereby at least 10 of the 17 ESs identified in T. brucei 427 can be functionally active in vitro. We could specifically select for switches mediated by DNA rearrangements by inducing VSG RNAi in the presence of drug selection for the active ES. Most of the preferentially activated VSGs could be activated by multiple mechanisms. This VSG RNAi‐based procedure provides a rapid and powerful means for analysing VSG switching in African trypanosomes entirely in vitro.
CesT is a multi‐effector chaperone and recruitment factor required for the efficient type III secretion of both LEE‐ and non‐LEE‐encoded effectors of enteropathogenic Escherichia coli
Authors: Thomas Nikhil A. , Deng Wanyin , Puente Jose L. , Frey Elizabeth A. , Yip Calvin K. , Strynadka Natalie C. J. , Finlay B. Brett
Published: August 2005

Enteropathogenic Escherichia coli (EPEC) is an intestinal attaching and effacing pathogen that utilizes a type III secretion system (T3SS) for the delivery of effectors into host cells. The chaperone CesT has been shown to bind and stabilize the type III translocated effectors Tir and Map in the bacterial cytoplasm prior to their delivery into host cells. In this study we demonstrate  a  role  for  CesT  in  effector  recruitment  to the membrane embedded T3SS. CesT‐mediated effector recruitment was dependent on the presence of the T3SS membrane‐associated ATPase EscN. EPEC ΔcesT carrying a C‐terminal CesT variant, CesT(E142G), exhibited normal cytoplasmic Tir stability function, but was less efficient in secreting Tir, further implicating CesT in type III secretion. In vivo co‐immunoprecipitation studies using CesT‐FLAG containing EPEC lysates demonstrated that CesT interacts with Tir and EscN, consistent with the notion of CesT recruiting Tir to the T3SS. CesT was also shown to be required for the efficient secretion of several type III effectors encoded within and outside the locus of enterocyte effacement (LEE) in addition to Tir and Map. Furthermore, a CesT affinity column was shown to specifically retain multiple effector proteins from EPEC culture supernatants. These findings indicate that CesT is centrally involved in recruiting multiple type III effectors to the T3SS via EscN for efficient secretion, and functionally redefine the role of CesT in multiple type III effector interactions.
The fungal hydrophobin RolA recruits polyesterase and laterally moves on hydrophobic surfaces
Authors: Takahashi Toru , Maeda Hiroshi , Yoneda Sachiyo , Ohtaki Shinsaku , Yamagata Yohei , Hasegawa Fumihiko , Gomi Katsuya , Nakajima Tasuku , Abe Keietsu
Published: August 2005

When fungi grow on plant or insect surfaces coated with wax polyesters that protect against pathogens, the fungi generally form aerial hyphae to contact the surfaces. Aerial structures such as hyphae and conidiophores are coated with hydrophobins, which are surface‐active proteins involved in adhesion to hydrophobic surfaces. When the industrial fungus Aspergillus oryzae was cultivated in a liquid medium containing the biodegradable polyester polybutylene succinate‐coadipate (PBSA), the rolA gene encoding hydrophobin RolA was highly transcribed. High levels of RolA and its localization on the cell surface in the presence of PBSA were confirmed by immunostaining. Under these conditions, A. oryzae simultaneously produced the cutinase CutL1, which hydrolyses PBSA. Pre‐incubation of PBSA with RolA stimulated PBSA degradation by CutL1, suggesting that RolA bound to the PBSA surface was required for the stimulation. Immunostaining revealed that PBSA films coated with RolA specifically adsorbed CutL1. Quartz crystal microbalance analyses further demonstrated that RolA attached to a hydrophobic sensor chip specifically adsorbed CutL1. Circular dichroism spectra of soluble‐state RolA and bound RolA suggested that RolA underwent a conformational change after its adsorption to hydrophobic surfaces. These results suggest that RolA adsorbed to the hydrophobic surface of PBSA recruits CutL1, resulting in condensation of CutL1 on the PBSA surface and consequent stimulation of PBSA hydrolysis. A fluorescence recovery after photobleaching experiment on PBSA films coated with FITC‐labelled RolA suggested that RolA moves laterally on the film. We discuss the novel molecular functions of RolA with regard to plastic degradation.
TrmB, a sugar sensing regulator of ABC transporter genes in Pyrococcus furiosus exhibits dual promoter specificity and is controlled by different inducers
Authors: Lee Sung‐Jae , Moulakakis Christina , Koning Sonja M. , Hausner Winfried , Thomm Michael , Boos Winfried
Published: August 2005

TrmB is the transcriptional repressor for the gene cluster of the trehalose/maltose ABC transporter of the hyperthermophilic archaea Thermococcus litoralis and Pyrococcus furiosus (malE or TM operon), with maltose and trehalose acting as inducers. We found that TrmB (the protein is identical in both organisms) also regulated the transcription of genes encoding a separate maltodextrin ABC transporter in P. furiosus (mdxE or MD operon) with maltotriose, longer maltodextrins and sucrose acting as inducers, but not with maltose or trehalose. In vitro transcription of the malE and the mdxE operons was inhibited by TrmB binding to the different operator sequences. Inhibition of the TM operon was released by maltose and trehalose whereas inhibition of the MD operon was released by maltotriose and larger maltodextrins as well as by sucrose. Scanning mutagenesis of the TM operator revealed the role of the palindromic TACTNNNAGTA sequence for TrmB recognition. TrmB exhibits a broad spectrum of sugar‐binding specificity, binding maltose, sucrose, maltotriose and trehalose in decreasing order of affinity, half‐maximal binding occurring at 20, 60, 250 and 500 µM substrate concentration respectively. Of all substrates, only maltose shows sigmoidal binding characteristics with a Hill coefficient of 2. As measured by molecular sieve chromatography and cross‐linking TrmB behaved as dimer in dilute buffer solution at room temperature. We conclude that TrmB acts as a bifunctional transcriptional regulator acting on two different promoters and being differentially controlled by binding to different sugars. We believe this to represent a novel strategy of prokaryotic transcription regulation.