bluuebunny/crossref_metadata_2025 huggingface dataset:
Epigenetics and T helper 1 differentiation
Authors: Aune Thomas M. , Collins Patrick L. , Chang Shaojing
Published: December 2008

Naïve T helper cells differentiate into two subsets, T helper 1 and 2, which either transcribe the Ifng gene and silence the Il4 gene or transcribe the Il4 gene and silence the Ifng gene, respectively. This process is an essential feature of the adaptive immune response to a pathogen and the development of long‐lasting immunity. The ‘histone code’ hypothesis proposes that formation of stable epigenetic histone marks at a gene locus that activate or repress transcription is essential for cell fate determinations, such as T helper 1/T helper 2 cell fate decisions. Activation and silencing of the Ifng gene are achieved through the creation of stable epigenetic histone marks spanning a region of genomic DNA over 20 times greater than the gene itself. Key transcription factors that drive the T helper 1 lineage decision, signal transducer and activator 4 (STAT4) and T‐box expressed in T cells (T‐bet), play direct roles in the formation of activating histone marks at the Ifng locus. Conversely, STAT6 and GATA binding protein 3, transcription factors essential for the T helper 2 cell lineage decision, establish repressive histone marks at the Ifng locus. Functional studies demonstrate that multiple genomic elements up to 50 kilobases from Ifng play critical roles in its proper transcriptional regulation. Studies of three‐dimensional chromatin conformation indicate that these distal regulatory elements may loop towards Ifng to regulate its transcription. We speculate that these complex mechanisms have evolved to tightly control levels of interferon‐γ production, given that too little or too much production would be very deleterious to the host.
Assessment of pulp vitality: a review
Authors: GOPIKRISHNA VELAYUTHAM , PRADEEP GALI , VENKATESHBABU NAGENDRABABU
Published: December 2008

Background.  One of the greatest diagnostic challenges in clinical practice is the accurate assessment of pulp status. This may be further complicated in paediatric dentistry where the practitioner is faced with a developing dentition, traumatized teeth, or young children who have a limited ability to recall a pain history for the tooth in question. A variety of pulp testing approaches exist, and there may be confusion as to their validity or appropriateness in different clinical situations.

Aim.  The aim of this paper is to provide the clinician with a comprehensive review of current pulp testing methods. A key objective is to highlight the difference between sensitivity testing and vitality testing. A biological basis for pulp testing is also provided to allow greater insight into the interpretation of pulp testing results. The rationale for, and methods of, assessing pulpal blood flow are described.
Histological comparison of pulpal inflammation in primary teeth with occlusal or proximal caries
Authors: KASSA DESPOINA , DAY PETER , HIGH ALEX , DUGGAL MONTY
Published: December 2008

Objective.  A number of clinical and histological studies have investigated caries‐related changes in the primary tooth pulp, but the effect of caries site, as a clinical variable, has not been previously considered. This study sought to compare inflammatory changes within the pulp of primary molars according to the location of the caries lesion (occlusal or proximal).

Methods.  Eighty‐three primary molars were extracted under general anaesthesia for caries and/or orthodontic reasons, and were split immediately after removal and fixed in 10% formalin. Teeth were then decalcified, sectioned, and stained with haematoxylin and eosin for histological examination using light microscopy. Caries depth was measured using a graticle, and the site of the caries lesion noted as occlusal or proximal. Samples were further classified into one of five subgroups according to the observed degree of pulpal inflammation.

Results.  Key findings were that where caries depth was less than 50% of the total dentine thickness, there were no significant differences in inflammatory status according to caries site. In contrast, marked inflammatory changes were significantly more likely throughout the coronal pulp of teeth with proximal caries compared to teeth with occlusal caries where caries depth was equal to, or greater than, 50% of the total dentine thickness (P = 0.017, Fisher's exact test).

Conclusion.  Primary teeth with proximal carious lesions extending more than 50% through the dentine thickness appear to have more extensive inflammatory pulpal changes than teeth with occlusal caries of a similar depth. This finding has clinical implications and may help inform treatment decisions in the management of primary teeth with deep carious lesions.
Pulpal status of human primary teeth with physiological root resorption
Authors: MONTEIRO JOANA , DAY PETER , DUGGAL MONTY , MORGAN CLAIRE , RODD HELEN
Published: December 2008

Objective.  The overall aim of this study was to determine whether any changes occur in the pulpal structure of human primary teeth in association with physiological root resorption.

Methods.  The experimental material comprised 64 sound primary molars, obtained from children requiring routine dental extractions under general anaesthesia. Pulp sections were processed for indirect immunofluorescence using combinations of: (i) protein gene product 9.5 (a general neuronal marker); (ii) leucocyte common antigen CD45 (a general immune cell marker); and (iii) Ulex europaeus I lectin (a marker of vascular endothelium). Image analysis was then used to determine the percentage area of staining for each label within both the pulp horn and mid‐coronal region. Following measurement of the greatest degree of root resorption in each sample, teeth were subdivided into three groups: those with physiological resorption involving less than one‐third, one‐third to two‐thirds, and more than two‐thirds of their root length.

Results.  Wide variation was evident between different tooth samples with some resorbed teeth showing marked changes in pulpal histology. Decreased innervation density, increased immune cell accumulation, and increased vascularity were evident in some teeth with advanced root resorption. Analysis of pooled data, however, did not reveal any significant differences in mean percentage area of staining for any of these variables according to the three root resorption subgroups (P > 0.05, analysis of variance on transformed data).

Conclusions.  This investigation has revealed some changes in pulpal status of human primary teeth with physiological root resorption. These were not, however, as profound as one may have anticipated. It is therefore speculated that teeth could retain the potential for sensation, healing, and repair until advanced stages of root resorption.
Dental pulp stem cells: what, where, how?
Authors: SLOAN ALASTAIR J. , WADDINGTON RACHEL J.
Published: December 2008

Introduction.  It is now accepted that progenitor/stem cells reside within the post‐natal dental pulp. Studies have identified several niches of multipotent mesenchymal progenitor cells, known as dental pulp stem cells, which have a high proliferative potential for self‐renewal. These progenitor stem cells are now recognized as being vital to the dentine regeneration process following injury. Understanding the nature of these progenitor/stem cell populations in the pulp is important in determining their potentialities and development of isolation or recruitment strategies for use in regeneration and tissue engineering. Characterization of these cells, and determination of their potentialities in terms of specificity of regenerative response, may help direct new clinical treatment modalities. Such novel treatments may involve controlled direct recruitment of the cells in situ and possible seeding of stem cells at sites of injury for regeneration or use of the stem cells with appropriate scaffolds for tissue engineering solutions. Such approaches may provide an innovative and novel biologically based new generation of clinical materials and/or treatments for dental disease.

Aim.  This study aimed to review the body of knowledge relating to stem cells and to consider the possibility of these cell populations, and related technology, in future clinical applications.
Perceptions of internal marketing and organizational commitment by nurses
Authors: Chang Ching Sheng , Chang Hae Ching
Published: December 2008

Title.  Perceptions of internal marketing and organizational commitment by nurses.

Aim.  This paper is a report of a study to determine whether a favourable perception of internal marketing is associated with increased organizational commitment.

Background.  The role of nurses in healthcare treatment is expanding, and becoming more important as time progresses. Therefore, the primary concern of business of health care is to use internal marketing strategies effectively to enhance and develop nurses’ organizational commitment and reduce turnover to promote competitive advantages for the organization.

Methods.  A cross‐sectional design was used. Questionnaires were distributed in 2006 to a convenience sample of 450 Registered Nurses in two teaching hospitals in Taiwan, and 318 questionnaires were returned. Eighteen were excluded because of incomplete answers, which left 300 usable questionnaires (response rate 66·7%). Validity and reliability testing of the questionnaire proved satisfactory and Structural Equation Modeling was used to analyse the data.

Results.  A favourable perception of internal marketing was associated with increased organizational commitment. Communication management had the greatest influence on organizational commitment and external activity had the smallest impact.

Conclusion.  Hospital managers need to recognize the importance of internal marketing for staff retention and the survival of their organizations as competitive pressure increases. As a great deal of time and costs are involved in educating nurses, the best way to retain outstanding nurses and reduce turnover costs and personnel problems is for employers to understand the needs and expectations of their nursing staff.
Beta diversity of frogs in the forests of New Guinea, Amazonia and Europe: contrasting tropical and temperate communities
Authors: Dahl Chris , Novotny Vojtech , Moravec Jiri , Richards Stephen J.
Published: December 2008

Aim  To test the hypothesis that animal communities within environmentally relatively uniform lowland forests are characterized by low beta diversity, both in tropical and in temperate areas.

Location  Lowland forests in the basins of the Sepik and Ramu rivers in New Guinea, the Amazon river in Bolivia, and the Elbe and Dyje rivers in the Czech Republic.

Methods  A network of 5–6 study sites spanning distances from 20–80 to 300–500 km in each study area was systematically surveyed for all frogs, using visual detection and call tracking. The community data were analysed for alpha and beta diversity.

Results  Local (alpha) diversity of frog communities was similar in the two tropical areas, New Guinea (mean ± SE of 22 ± 1.4 species per site) and Amazonia (24 ± 1.7 species), but was significantly lower in Europe (8 ± 0.8 species). In Amazonia, 36 of the total of 70 species were recorded from single sites. In contrast, widespread species dominated in Europe, whereas New Guinea exhibited an intermediate pattern with both local and widespread species well represented. The rate of species accumulation across different sites was lowest in Europe, intermediate in New Guinea and highest in Amazonia. The regional species diversity, expressed as the combined number of species from five study sites, was 1.5 times higher than the local species diversity at a single site in Europe, 2.0 times higher in New Guinea and 2.7 times higher in Amazonia. The proportion of species shared between communities decreased with geographic distance in New Guinea and Europe, but not in Amazonia.

Main conclusions  Frog communities in the lowland tropical rain forests of New Guinea and Amazonia had similar numbers of species, but differed in their beta diversity. More species in Amazonia had restricted distributions than in New Guinea. Both tropical areas had markedly higher alpha and beta diversity than the temperate area in Europe.
Myelodysplastic syndromes: biology and treatment
Authors: Jädersten M. , Hellström‐Lindberg E.
Published: December 2008

Optimal management of patients with myelodysplastic syndromes (MDS) requires an insight into the biology of the disease and the mechanisms of action of the available therapies. This review focuses on low‐risk MDS, for which chronic anaemia and eventual progression to acute myeloid leukaemia are the main concerns. We cover the updated World Health Organization classification, the latest prognostic scoring system, and describe novel findings in the pathogenesis of 5q− syndrome. We perform in depth analyses of two of the most widely used treatments, erythropoietin and lenalidomide, discussing mechanisms of action, reasons for treatment failure and influence on survival.
The sodium‐dependentd‐glucose transport protein ofHelicobacter pylori
Authors: Psakis Georgios , Saidijam Massoud , Shibayama Keigo , Polaczek Julia , Bettaney Kim E. , Baldwin Jocelyn M. , Baldwin Stephen A. , Hope Ryan , Essen Lars‐Oliver , Essenberg Richard C. , Henderson Peter J. F.
Published: December 2008

Helicobacter pyloriis a Gram‐negative pathogenic microaerophile with a particular tropism for the mucosal surface of the gastric epithelium. Despite its obligatory microaerophilic character, it can metabolized‐glucose and/ord‐galactose in both oxidative and fermentative pathways via a Na+‐dependent secondary active transport, a glucokinase and enzymes of the pentose phosphate pathway. We have assigned the Na+‐dependent transport of glucose to the protein product of theH. pylori 1174gene. The gene was heterologously expressed in a glucose transport‐deficientEscherichia colistrain, where transport activities of radiolabelledd‐glucose,d‐galactose and 2‐deoxy‐d‐glucose were restored, consistent with the expected specificity of the hexose uptake system inH. pylori.d‐Mannose was also identified as a substrate. The HP1174 transport protein was purified and reconstituted into proteoliposomes, where sodium dependence of sugar transport activity was demonstrated. Additionally the tryptophan/tyrosine fluorescence of the purified protein showed quenching by 2‐deoxy‐d‐glucose,d‐mannose,d‐glucose ord‐galactose in the presence of sodium ions. This is the first reported purification and characterization of an active glucose transport protein member of the TC 2.1.7 subgroup of the Major Facilitator Superfamily, constituting the route for entry of sugar nutrients intoH. pylori. A model is derived of its three‐dimensional structure as a paradigm of the family.
Sodium‐dependent dynamic assembly of membrane complexes in sodium‐driven flagellar motors
Authors: Fukuoka Hajime , Wada Tomoyuki , Kojima Seiji , Ishijima Akihiko , Homma Michio
Published: December 2008

The bacterial flagellar motor is driven by the electrochemical potential of specific ions, H+ or Na+. The motor consists of a rotor and stator, and their interaction generates rotation. The stator, which is composed of PomA and PomB in the Na+ motor of Vibrio alginolyticus, is thought to be a torque generator converting the energy of ion flux into mechanical power. We found that specific mutations in PomB, including D24N, F33C and S248F, which caused motility defects, affected the assembly of stator complexes into the polar flagellar motor using green fluorescent protein‐fused stator proteins. D24 of PomB is the predicted Na+‐binding site. Furthermore, we demonstrated that the coupling ion, Na+, is required for stator assembly and that phenamil (an inhibitor of the Na+‐driven motor) inhibited the assembly. Carbonyl cyanide m‐chlorophenylhydrazone, which is a proton ionophore that collapses the sodium motive force in this organism at neutral pH, also inhibited the assembly. Thus we conclude that the process of Na+ influx through the channel, including Na+ binding, is essential for the assembly of the stator complex to the flagellar motor as well as for torque generation.
Ss‐LrpB, a transcriptional regulator from Sulfolobus solfataricus, regulates a gene cluster with a pyruvate ferredoxin oxidoreductase‐encoding operon and permease genes
Authors: Peeters Eveline , Albers Sonja‐Verena , Vassart Amelia , Driessen Arnold J. M. , Charlier Daniel
Published: December 2008

Ss‐LrpB is an Lrp‐like transcriptional regulator from Sulfolobus solfataricus. Previously, in vitro binding of Ss‐LrpB to the control region of its own gene has been extensively studied. However, nothing was known about the physiological role of this regulator yet. Here, using the knowledge of the DNA‐binding sequence specificity of Ss‐LrpB, several potential binding sites were predicted in silico in promoter regions of genes located adjacently to the Ss‐lrpB gene. These genes include an operon encoding a pyruvate ferredoxin oxidoreductase (porDAB) and two genes encoding putative permeases. In vitro protein–DNA interaction studies allowed the identification of the Ss‐LrpB binding sites in the cognate control regions. Intriguingly, the binding site organization in the por operator is identical to that in the Ss‐lrpB control region. An Ss‐lrpB gene disruption mutant was constructed and the gene expression of the above‐mentioned targets in this mutant was analysed by qRT‐PCR and compared with isogenic wild type. Our data demonstrate that in vivo Ss‐LrpB acts as an activator at the promoters of the three predicted targets. Based on these results, it appears that not all regulators belonging to the archaeal Lrp family perform a function related to the amino acid metabolism, unlike the bacterial Lrp‐like regulators.
Atypical association of DDE transposition with conjugation specifies a new family of mobile elements
Authors: Brochet Mathieu , Da Cunha Violette , Couvé Elisabeth , Rusniok Christophe , Trieu‐Cuot Patrick , Glaser Philippe
Published: December 2008

We describe in Streptococcus agalactiae an atypical family of conjugative transposons named TnGBSs which associates DDE transposition and conjugation. We present evidence that the transposition of TnGBS2, the prototype of this family, is catalysed by a new class of DDE transposases that are widespread in Gram‐positive bacteria. Remarkably, transposition occurs in intergenic regions, 15 or 16 bp upstream the −35 sequence of promoters, minimizing the burden on the host cell and suggesting an association between transcription and transposition. Transposition catalyses the formation of a circular intermediate that is substrate for subsequent conjugative intercellular transfer. Conjugation is initiated at an origin of transfer by a transposon‐encoded relaxase. Whereas all integrative and conjugative elements described so far encode a phage‐related integrase, TnGBS2 is the first example of conjugative transposon whose recombination is mediated by a DDE transposase. The combination of DDE transposition with conjugation implies recombination constraints linked to the physical separation of donor and recipient molecules.
Does magnetic stimulation of sacral nerve roots modify colonic motility? Results of a randomized double‐blind sham‐controlled study
Authors: Gallas S. , Gourcerol G. , Ducrotté P. , Mosni G. , Menard J.‐f. , Michot F. , Leroi A.‐m.
Published: December 2008

Abstract  Although sacral nerve root stimulation (SNS) can result in a symptomatic improvement of faecal incontinence, the mechanism of action remains unknown. The aim of this study was to assess whether short‐term magnetic SNS can inhibit pharmacologically induced propulsive colonic contractions. Twelve healthy volunteers (median age: 43.5 years old) were studied on two separate occasions and randomized into either active (15 Hz, 100% output intensity for 5 s min−1 for 30 min) or sham rapid rate lumbosacral magnetic stimulation (rLSMS). Colorectal motility was recorded with a manometric catheter located at the most proximal transducer in the left colon and the most distal, in the rectum. Colonic contractions were provoked by instilling Bisacodyl. The effects of rLSMS on colonic, sigmoid and rectal contractions were monitored and recorded after Bisacodyl instillation. The appearance of high‐amplitude contractions propagated or not (HAC/HAPC) provoked by Bisacodyl instillation was significantly delayed during active compared to sham stimulation (P = 0.03). There was no difference in the characteristics of HAC/HAPC (i.e. frequency, amplitude, duration, velocity of propagation) or the motility index with active or sham stimulation. The perception of urgency tended to be decreased with rLSMS following Bisacodyl instillation. The catheter was expulsed within a median of 16.5 min (range 8–39) after Bisacodyl administration during active stimulation compared to 14 min (range 5–40) during sham stimulation (P = 0.03). This study suggests that rLSMS could delay the appearance of the first Bisacodyl‐induced colonic contractions.
Dual control of avirulence in Leptosphaeria maculans towards a Brassica napus cultivar with ‘sylvestris‐derived’ resistance suggests involvement of two resistance genes
Authors: Van de Wouw A. P. , Marcroft S. J. , Barbetti M. J. , Hua Li , Salisbury P. A. , Gout L. , Rouxel T. , Howlett B. J. , Balesdent M. H.
Published: December 2008

Blackleg disease (phoma stem canker) of Brassica napus (canola, oilseed rape) is caused by the fungus Leptosphaeria maculans. In some regions of Australia, resistance in oilseed rape cultivars derived from B. rapa subs. sylvestris (e.g. cv. Surpass 400) became ineffective within three years of commercial release. The genetic control of avirulence in L. maculans towards cv. Surpass 400 is described. When Australian field isolates were screened on this cultivar, three phenotypic classes were observed; virulent, intermediate and avirulent. Analysis of crosses between fungal isolates varying in their ability to infect cv. Surpass 400 demonstrated the presence of two unlinked avirulence genes, AvrLm1 and AvrLmS. Complementation of isolates (genotype avrLm1) with a functional copy of AvrLm1, and genotyping of field isolates using a molecular marker for AvrLm1 showed that virulence towards Rlm1 is necessary, but not sufficient, for expression of a virulent phenotype on cv. Surpass 400. Taken together, these data strongly suggest that cv. Surpass 400, with ‘sylvestris‐derived’ resistance, contains at least two resistance genes, one of which is Rlm1.
Induction of Multiepitopic and Long‐Lasting Immune Responses Against Tumour Antigens by Immunization with Peptides, DNA and Recombinant Adenoviruses Expressing Minigenes
Authors: Durántez M. , López‐Vázquez A. B. , De Cerio A. L.‐D. , Huarte E. , Casares N. , Prieto J. , Borrás‐Cuesta F. , Lasarte J. J. , Sarobe P.
Published: December 2008

The development of immunization strategies to induce strong and multiepitopic T‐cell responses against tumour antigens is needed for anti‐tumour immunotherapy. However, a common finding after immunization with complex antigens is the preferential induction of immune responses against immunodominant epitopes. In this study, with the aim of inducing multiepitopic responses against several common tumour antigens, we have designed a minigene construct encoding four human leucocyte antigen (HLA)‐A2‐restricted epitopes belonging to tumour antigens CEA (CEA‐691 and CEA‐571), MAGE2 (MAGE2‐157) and MAGE3 (MAGE3‐112), as well as the universal PADRE epitope recognized by T helper lymphocytes. To optimize the activation of immune responses against these epitopes, we have used different antigen formats (short peptides encompassing individual epitopes and DNA plasmids or adenoviral constructs expressing the minigene) in single or combined immunization schedules. A single immunization with either DNA plasmid or recombinant adenovirus induced a monospecific immune response against the immunodominant epitope CEA‐571, whereas immunization with the peptide pool induced responses against all epitopes. Combination of peptide priming followed by a boost with the plasmid and the recombinant adenovirus expressing the minigene induced stronger, multi‐specific and long‐lasting immune responses, overcoming the immunodominance imposed by the main T‐cell epitope. Moreover, these combined immunization strategies were able to induce responses that were able to recognize Mel624 HLA‐A2+ tumour cells expressing MAGE2. These results suggest that heterologous immunization strategies combining peptides and DNA or recombinant adenoviruses can be useful to broaden the specificity and enhance the efficacy of subunit vaccines.
Arabidopsis MYB30 is a direct target of BES1 and cooperates with BES1 to regulate brassinosteroid‐induced gene expression
Authors: Li Lei , Yu Xiaofei , Thompson Addie , Guo Michelle , Yoshida Shigeo , Asami Tadao , Chory Joanne , Yin Yanhai
Published: December 2008

A paradox of plant hormone biology is how a single small molecule can affect a diverse array of growth and developmental processes. For instance, brassinosteroids (BRs) regulate cell elongation, vascular differentiation, senescence and stress responses. BRs signal through the BES1/BZR1 (bri1‐Ethylmethane Sulphonate suppressor 1/brassinazole‐resistant 1) family of transcription factors, which regulate hundreds of target genes involved in this pathway, yet little is known of this transcriptional network. Through microarray and chromatin immunoprecipitation (ChIP) experiments, we identified a direct target gene of BES1, AtMYB30, which encodes an MYB family transcription factor. AtMYB30 null mutants display decreased BR responses and enhance the dwarf phenotype of a weak allele of the BR receptor mutant bri1. Many BR‐regulated genes have reduced expression and/or hormone‐induction in AtMYB30 mutants, indicating that AtMYB30 functions to promote expression of a subset of BR target genes. AtMYB30 and BES1 bind to a conserved MYB‐binding site and E‐box sequences, respectively, in the promoters of genes that are regulated by both BRs and AtMYB30. Finally, AtMYB30 and BES1 interact with each other both in vitro and in vivo. These results demonstrate that BES1 and AtMYB30 function cooperatively to promote BR target gene expression. Our results therefore establish a new mechanism by which AtMYB30, a direct target of BES1, functions to amplify BR signaling by helping BES1 activate downstream target genes.
The chloroplast protein RPH1 plays a role in the immune response of Arabidopsis to Phytophthora brassicae
Authors: Belhaj Khaoula , Lin Baiqing , Mauch Felix
Published: December 2008

Plant immune responses to pathogens are often associated with enhanced production of reactive oxygen species (ROS), known as the oxidative burst, and with rapid hypersensitive host cell death (the hypersensitive response, HR) at sites of attempted infection. It is generally accepted that the oxidative burst acts as a promotive signal for HR, and that HR is highly correlated with efficient disease resistance. We have identified the Arabidopsis mutant rph1 (resistance to Phytophthora 1), which is susceptible to the oomycete pathogen Phytophthora brassicae despite rapid induction of HR. The susceptibility of rph1 was specific for P. brassicae and coincided with a reduced oxidative burst, a runaway cell‐death response, and failure to properly activate the expression of defence‐related genes. From these results, we conclude that, in the immune response to P. brassicae, (i) HR is not sufficient to stop the pathogen, (ii) HR initiation can occur in the absence of a major oxidative burst, (iii) the oxidative burst plays a role in limiting the spread of cell death, and (iv) RPH1 is a positive regulator of the P. brassicae‐induced oxidative burst and enhanced expression of defence‐related genes. Surprisingly, RPH1 encodes an evolutionary highly conserved chloroplast protein, indicating a function of this organelle in activation of a subset of immune reactions in response to P. brassicae. The disease resistance‐related role of RPH1 was not limited to the Arabidopsis model system. Silencing of the potato homolog StRPH1 in a resistant potato cultivar caused susceptibility to the late blight pathogen Phytophthora infestans.
The James Blundell Award Lecture 2007: Do we really understand immune red cell destruction?
Authors: Garratty G.
Published: December 2008

summary We have learned a great deal about immune red blood cell (RBC) destruction since the elaboration of biochemical/immunological interactions of antibodies, complement and macrophages during the past 50 years. We first learned about the direct lysis of RBCs involving complement. We then learned of the role of the macrophage (particularly in the spleen and the liver) in initiating phagocytosis and antibody‐dependent cytotoxicity of antibody‐coated RBCs. Later, as the complexities of the human complement system were unravelled, we learned that complement‐coated RBCs that were not directly haemolysed could interact with macrophages and that specific complement molecules on the RBC membrane could lead to a phagocytic event or the RBC (although heavily coated with complement) could survive normally. The application of isotope‐labelling procedures (e.g. 51Cr) for RBC survival (starting in the 1950s) advanced our knowledge considerably. Advances in knowledge in immunology helped us understand the complexity of the immunoglobulins (e.g. subclasses) and the specific receptors on macrophages and their role in immune haemolysis. Nevertheless, after more than 30 years researching this area, I am sometimes embarrassed to realize how much I cannot explain. Why do some patients have severe haemolytic transfusion reactions because of antibodies that are only detectable by one technique or not detectable by any? How do we explain autoimmune haemolytic anaemia with negative direct antiglobulin tests (DATs)? Why do RBCs strongly coated with immunoglobulin (Ig)G1 or IgG3 sometimes have normal survival? Are cells, other than macrophages, involved in immune RBC destruction? Could the relative amount of cytotoxicity vs. phagocytosis explain different clinical findings and response to treatment? How do we explain ‘hyperhaemolysis’ in sickle cell disease? Could novel mechanisms involving IgG glycosylation, CD47, ‘armed’ macrophages, bystander lysis, antibody activated reactive oxygen species, natural killer cells or antibody perturbation of RBC membrane be involved? Why do RBCs die after circulating for 100–120 days in healthy individuals? How should we define a ‘clinically significant’ antibody; how do we evaluate this? So many questions, so little time!
Erythrocyte ageingin vivoandin vitro: structural aspects and implications for transfusion
Authors: Bosman G. J. C. G. M. , Werre J. M. , Willekens F. L. A. , Novotný V. M. J.
Published: December 2008

summaryErythrocyte transfusion is essential in conditions of large blood loss, of inadequate bone marrow production and of increased erythrocyte breakdown. The structural and biochemical changes that erythrocytes go through during storage, probably associated with the disappearance of up to 30% of the erythrocytes within 24 h after transfusion, are likely to contribute to the transfusion side effects: iron overload, erythrocyte adhesion to the endothelial surface with proinflammatory consequences, autoantibody formation, endothelial damage by released erythrocyte constituents, a hampered microcirculation and oxygen delivery.In vivo, senescent erythrocytes are marked for removal by binding of autologous immunoglobulin G to ageing antigens, which arise by changes in the conformation of the membrane domain of band 3. Also, vesicle formation has been described as an integral part of the erythrocyte ageing process. Comparable changes occur during erythrocyte storage. This review describes the current state of knowledge of the mechanism of erythrocyte ageingin vivo, ageing‐related changes occurring during erythrocyte storage in blood bank conditions and their possible relation with the transfusion side effects. In view of the key position of band 3 in the maintenance of erythrocyte structure and function, elucidation of the pathways that control posttranslational modification of band 3 during storage may lead to new approaches towards maintaining ATP concentration and cellular integrity. This review concludes with the challenge to further explore the underlying processes of erythrocyte ageing in order to provide physiologically relevant tools for assessing and predicting erythrocyte homeostasisin vitroandin vivoand thereby to contribute to the development of rational transfusion protocols for various patient categories.
Hispanic ethnicity, race and blood donation in the United States
Authors: Gillum F. , Eder A. F. , McLaurin‐Jones T. L.
Published: December 2008

summary The aim of this study was to assess the hypothesis that blood donation rates vary with Hispanic ethnicity (family origin in Spanish‐speaking countries) in addition to race in the United States. Lower blood donation rates have been reported among African Americans (AAs) compared with non‐Hispanic European Americans (EAs). Adequate published reports on donation rates are not available for Hispanic Americans (HAs). Using data from a 2002 national survey, which included 4923 men and 7600 women aged 15–44 years with complete data, we tested the hypothesis using weighted bivariate and multivariate statistics. Among men aged 25–44 years, the percentage [95% confidence limits (95% CL)] with a history of blood donation since 1985 was similar at ages 25–34 years (46%, 42–49) and 35–44 years (41%, 37–45). It was highest in non‐Hispanic EA (49%, 45–52%), intermediate in AA (35%, 30–40%) and lowest in HA (30%, 25–36%) (P < 0·001). Other variables significantly (P < 0·01) associated with history of blood donation in bivariate analyses were nativity (United States/other), education (<12/≥12 years), poverty (<200%/≥200% poverty limit) and married (yes/no). Variables that are not significantly associated were age, metropolitan residence (yes/no), receipt of public assistance (yes/no), current labour‐force participation (yes/no) and religion raised. Compared with non‐Hispanic EA, the adjusted odds ratios were essentially the same for Hispanics 0·66 (95% CL 0·47–0·92) and AAs 0·64 (95% CL 0·49–0·84). Only 34% of women had donated blood, but the association with race/ethnicity was similar. Similar patterns were also seen at ages 15–24 years. HAs and AAs have similar low blood donation rates compared with non‐Hispanic EAs. The difference is not explained by sociodemographic variables.
Spore‐forming organisms in platelet concentrates: a challenge in transfusion bacterial safety
Authors: Störmer M. , Vollmer T. , Kleesiek K. , Dreier J.
Published: December 2008

summary Bacterial detection and pathogen reduction are widely used methods of minimizing the risk of transfusion‐transmitted bacterial infection. But, bacterial spores are highly resistant to chemical and physical agents. In this study, we assessed the bacterial proliferation of spore‐forming organisms seeded into platelet concentrates (PCs) to demonstrate that spores can enter the vegetative state in PCs during storage. In the in vitro study, PCs were inoculated with 1–10 spores mL−1of Bacillus cereus (n = 1), Bacillus subtilis (n = 2) and Clostridium sporogenes (n = 2). Sampling was performed during 6‐day aerobic storage at 22 °C. The presence of bacteria was assessed by plating culture, automated culture and real‐time reverse transcriptase–polymerase chain reaction (RT‐PCR). Spores of the C. sporogenes do not enter the vegetative phase under PC storage conditions, whereas B. subtilis and B. cereus showed growth in the PC and could be detected using RT‐PCR and automated culture. Depending on the species and inoculums, bacterial spores may enter the vegetative phase during PC storage and can be detected by bacterial detection methods.
Free distribution of insecticide treated bed nets to pregnant women in Kinshasa: an effective way to achieve 80% use by women and their newborns
Authors: Pettifor Audrey , Taylor Eboni , Nku David , Duvall Sandra , Tabala Martine , Mwandagalirwa Kashamuka , Meshnick Steve , Behets Frieda
Published: December 2008

Objective  To determine whether long lasting insecticide treated bed nets (LLINs) distributed free of charge to pregnant women at their first antenatal clinic visit in Kinshasa, DRC are used from the time of distribution to delivery and 6 months after delivery.

Methods  Women were enrolled into a cohort study at their first antenatal care (ANC) visit and provided LLINs free of charge. Reported use of these nets was then measured at the time of delivery (n = 328) and in a random sample of women (n = 100) 6 months post‐delivery using an interviewer administered, structured questionnaire.

Results  At baseline, only 25% of women reported having slept under a bed net the night before the interview. At the time of delivery, after being provided an LLIN for free, this increased to 79%. Six months post‐delivery (n = 100), 80% of women reported sleeping under a net with a child under the age of 5 the night before the interview.

Conclusions  Freely distributed bed nets are acceptable, feasible and result in high usage. Free distribution of bed nets during antenatal clinic visits may be a highly effective way to rapidly increase the use of bed nets among both pregnant women and their newborn infants in areas with high levels of ANC attendance.
The role of eosinophil major basic protein in angiogenesis
Authors: Puxeddu I. , Berkman N. , Nissim Ben Efraim A. H. , Davies D. E. , Ribatti D. , Gleich G. J. , Levi‐Schaffer F.
Published: December 2008

Background:  Eosinophil‐derived major basic protein (MBP) plays an active role in allergic inflammation and tissue remodelling. However, its role in angiogenesis has not been established as yet. Therefore our objective was to investigate whether MBP exhibits any direct pro‐angiogenic effects.

Methods:  Rat aortic endothelial cells and human umbilical vascular endothelial cells were cultured with different concentrations of MBP and their viability (Trypan blue exclusion test), proliferation (thymidine incorporation) and capillary‐like structure formation (matrigel assay) were investigated in vitro. The angiogenic activity of MBP was then tested in vivo using the chick chorio allantoic membrane (CAM) assay.

Results:  Subcytotoxic concentrations of MBP induce endothelial cell proliferation and enhance the pro‐mitogenic effect of vascular endothelial growth factor (VEGF), but do not affect their VEGF release. MBP promotes capillarogenesis by endothelial cells seeded on matrigel and sprouting formation in the CAM assay. Furthermore, we have shown that the pro‐angiogenic effect of MBP is not due to its cationic charge since stimulation of the CAMs with the synthetic polycation, poly‐l‐arginine does not induce any angiogenic effects.

Conclusions:  These data demonstrate that MBP has pro‐angiogenic effects in vitro and in vivo, providing a novel mechanism whereby MBP can participate in tissue inflammation and remodelling in atopic diseases.
Low toxicity of a conditioning with 8‐Gy total body irradiation, fludarabine and cyclophosphamide as preparative regimen for allogeneic hematopoietic stem cell transplantation in pediatric hematological malignancies
Authors: Yanagisawa Ryu , Nakazawa Yozo , Sakashita Kazuo , Tanaka Miyuki , Shikama Naoto , Kamijo Takehiko , Shiohara Masaaki , Koike Kenichi
Published: December 2008

Abstract:  We here report the efficacy and toxicity of a conditioning regimen with fractionated 8‐Gy TBI, fludarabine, and cyclophosphamide in allogeneic HSCT for pediatric hematological malignancies. Among 22 children who received related or unrelated HSCT, nine were transplanted with refractory disease and/or from HLA two or more loci‐mismatched family donors. None of the patients developed graft failure. The Seattle grading system revealed that 18 patients had no RRT, and the remaining patients had grade I gastrointestinal toxicity alone. The estimated overall survival and leukemia‐free survival at two yr were 57.1% and 48.0%, respectively, in 10 patients with acute lymphoblastic leukemia; 91.7% and 71.3%, respectively, in 12 patients with myeloid leukemia. The incidence of TRM was 4.8% at two yr. The rates of RRT above grade II and TRM in an 8‐Gy TBI‐containing regimen were significantly lower than the data of historical control patients who underwent 12‐Gy TBI and cyclophosphamide with or without etoposide. The intermediate‐dose TBI‐based conditioning regimen may confer successful engraftment combined with minimized RRT, although its efficacy should be further evaluated.
Revisited impact of recipient age on the outcome of living donor liver transplantation for biliary atresia in the recent “transplantation era” in Japan
Authors: Ohya Yuki , Okajima Hideaki , Nishimori Aya , Lee Kwang‐Jong , Shirouzu Yasumasa , Yamamoto Hidekazu , Takeichi Takayuki , Asonuma Katsuhiro , Inomata Yukihiro
Published: December 2008

Abstract:  To re‐evaluate the impact of recipient age on the outcome of LDLT for BA in an era in which LDLT is the established treatment for BA in Japan. Thirty‐one patients with BA who underwent LDLT were divided into four groups regarding the age at LDLT: infants <1 yr old (group A; n = 14); young children 1 to 6 yr old (group B; n = 8); school children 6 to 15 yr old (group B; n = 5); and adults ≥15 yr old (group D; n = 4). Pre‐, peri‐, and postoperative factors were compared among the four groups. There was no significant difference in number of the previous laparotomy among the groups. Cholestasis was the dominant indication in group A. PELD score in group B was lower than that in the other groups, and blood loss in group B was significantly less than in groups A and D. Ratio of the graft weight to the recipient’s body weight (GRWR) in group A was significantly higher than in other groups. Duration of operation in group D was lower than in groups A and B, but there was no significant difference in the length of postoperative hospital stay and graft survival. Although the case volume was not big, the age of the recipient did not have any significant impact on the outcome of LDLT in our series.
Sustainability of humoral responses to varicella vaccine in pediatric transplant recipients following a pretransplantation immunization strategy
Authors: Barton Michelle , Wasfy Samia , Melbourne Toni , Hébert Diane , Moore Dorothy , Robinson Joan , Marchese Rocio D. , Allen Upton D.
Published: December 2008

Abstract:  Varicella infections pose serious challenges for organ transplant recipients. To determine the safety and immunogenicity of the OMVV and determine the maintenance of OMVV responses in transplanted subjects at varying periods of immunosuppression within the first two yr following transplantation. Eligible subjects given a two‐dose OMVV pretransplantation were monitored for AE. Antibody levels were assessed at baseline, six wk post‐OMVV, pretransplantation and up to 24 months post‐transplantation. Seroprotection was defined as ≥5 gpEU. Twenty‐one seronegative children were vaccinated. Following 42 doses, no vaccine‐related serious AE occurred. Mab_titer were 17.8 (5.7–910.2) and 183.5 EU (18.8–8116.4) at six and 12 wk, respectively (p < 0.0001). Fourteen (66.7%) participants were transplanted at a median of 16 months (1.5–56) following OMVV and had Mab_titer of 27.2 EU (9.0–236.2) just prior to transplantation. Of 11 who had post‐transplantation serology, seroprotection was sustained at three, six and 12 months post‐transplantation in 10/11, 12/12 and 8/10 subjects. In five of six subjects with two‐yr follow‐up, antibody levels remained seroprotective. No breakthrough varicella infections occurred. The receipt of OMVV prior to transplantation induced humoral responses which persisted in the early months following transplantation and up to two yr post‐transplantation and was not associated with any serious adverse consequences.
Differential human serum‐mediated neutralization of PERV released from pig cells transfected with variants of hDAF
Authors: Okura Eiji , Ishimaru Aiko , Yamamoto Aki , Nakatsu Shino , Shirakura Ryota , Okabe Masaru , Sawa Yoshiki , Fukuzawa Masahiro , Okumura Meinoshin , Miyagawa Shuji
Published: December 2008

Abstract:  Background:  Expression of complement regulatory proteins (CRP) on pig endothelial cells (PEC) is an effective means of avoiding induction of hyperacute rejection by human sera. However, pig endogenous retrovirus (PERV) from PEC transfected with CRP may acquire resistance to human sera. This study investigated a form of transfected CRP that is easily expressed on PERV particles.

Methods:  The PEC line was transfected with the Lac Z gene and PERV‐B to investigate PERV infectivity using a Lac Z pseudo‐type assay. The cDNAs of several modified DAF (CD55) were then transfected into the PEC(Lac Z)/P‐B lines using lipofection. DAF expression was verified by FACS analysis. Complement‐dependent PEC lysis was tested to verify the complement regulatory function of the expressed DAF. HEK293 cells were incubated with PEC culture supernatants with or without human sera. The inoculated 293 cells were histochemically stained and Lac Z‐positive blue foci were counted. The rate of reduction in Lac Z‐positive cells resulting from the addition of human serum was then calculated. In addition, to assess the localization of the expressed DAF, flotation sucrose density analysis was performed.

Results:  While PERV released from PEC expressing delta‐short consensus repeat 2 (delta‐SCR2) DAF (lacking CRP function) showed no change in resistance to human serum compared to control cells, PERV from cells expressing delta‐SCR1 DAF (with CRP function) showed a significant increase in resistance. The DAF‐blocking antibody assay indicated that PERV from the DAF transfectants expressed DAF molecules on the surface of the retrovirus. While delta‐SCR1 DAF (PI‐anchor form) significantly inhibited the reduction of Lac Z‐positive cells by human serum, the reduction of Lac Z‐positive cells by human serum was less inhibited in the case of transmembrane (TM)‐types of DAF–HLA‐G, modified influenza hemagglutinin (HA) and MCP (delta‐CYT form). However, the reduction in each TM‐type DAF was slightly less than that observed in naive and mock cells. The flotation sucrose density analysis of these transfectants indicated that the PI‐anchor form of DAF is a raft‐associated protein, and most TM‐types of DAF are non‐raft proteins.

Conclusion:  Induction of resistance to human serum in PERV, depends on the form of the CRP tail. The CRP/TM hybrid that does not associate with lipid rafts, is a suitable form of CRP for gene transduction.