bluuebunny/crossref_metadata_2025 huggingface dataset:
Direct interaction between the Lu/B‐CAM adhesion glycoproteins and erythroid spectrin†
Authors: Kroviarski Yolande , El Nemer Wassim , Gane Pierre , Rahuel Cécile , Gauthier Emilie , Lecomte Marie Christine , Cartron Jean Pierre , Colin Yves , Le Van Kim Caroline
Published: July 2004

Lutheran (Lu) and Lu(v13), two glycoprotein (gp) isoforms belonging to the immunoglobulin superfamily, represent adhesion molecules that act as erythrocyte receptors for laminin 10/11. These two gps, which differ only by the length of their cytoplasmic tail, carry both Lu blood group and Basal Cell Adhesion Molecule (B‐CAM) antigens. Here, analysis of the Triton extractability of recombinant Lu and Lu(v13) gps in K562 transfected cells showed that both gps were mainly associated with the detergent‐insoluble material. Patching experiments using Cholera Toxin subunit B indicated that Lu gps were not localized in lipid rafts. Glutathione‐S‐transferase capture assays showed that the cytoplasmic domain of Lu and Lu(v13) bound to erythroid spectrin, present in a low ionic strength extract from red cell ghosts. Direct interaction with spectrin was confirmed by plasmon resonance assays. Site‐directed mutagenesis mapped a major interaction site with spectrin to the RK573‐574 motif, located on the cytoplasmic tail of Lu gp, in close vicinity to the inner leaflet of the membrane lipid bilayer. The two Lu adhesion gps represent the first example of a direct link between transmembrane proteins and spectrin in red blood cells. Since Lu gps are low abundant proteins, we speculate that their interaction with spectrin might be critical for signalling and receptor function rather than for participating in the linkage of the lipid bilayer to the red cell skeleton.
Impact of pre‐analytical handling on bone marrow mRNA gene expression
Authors: Breit Stephen , Nees Matthias , Schaefer Ulrike , Pfoersich Margit , Hagemeier Christian , Muckenthaler Martina , Kulozik Andreas E.
Published: July 2004

Large clinical trials on leukaemia, require the transport of bone marrow (BM) from participating clinics to central diagnostic laboratories. We have investigated the impact of RNA extraction protocols and time delays between sample aspiration and RNA extraction on RNA quality and gene expression profiles. Intact RNA can be extracted from BM samples stored at room temperature for up to 48 h. Gene expression analyses using Affymetrix U95Av2 GeneChipsTM and a custom‐designed cDNA array in parallel showed that even short‐term storage of BM has dramatic effects on mRNA expression of individual transcripts. Many probe sets/genes showed either reproducible deregulation (18·8%, analysis of variance <0·05), or inconsistent expression that differed from patient to patient (38·4%). Moderate alterations were observed in 42·8% genes, with a maximum fold change <2·0 in all experiments and at all time points. These profound effects complicate the use of unstabilized, shipped BM samples for gene expression analyses. The comparison of a variety of RNA stabilization reagents (e.g. PAXgene) resulted in partial conservation of the mRNA expression patterns. Immediate density centrifugation or erythrocyte lysis and freezing at −80°C represent simple procedures that reliably preserved mRNA gene expression patterns in BM.
Applications of murine and humanized chimaeric monoclonal antibodies for red cell phenotyping
Authors: Lee E. , Burgess G. , Halverson G. R. , Huang T. J. , Reid M. E.
Published: July 2004

The limited supply of reagent human polyclonal antibodies to high prevalence antigens, like Jsb, is driving the search for alternative reagents. Murine immunoglobulin G (IgG) monoclonal antibodies (Mabs) and their humanized chimaeric IgM isoforms can now be used for typing patients and screening donors. Antigen typing of red blood cells (RBC) with a positive direct antiglobulin test (DAT) is also possible using these antibodies. Blood from patients with sickle cell disease and African donors were tested with reagent anti‐Jsb, murine Mab IgG anti‐Jsb [murine immunochemistry monoclonal antibody‐8 (MIMA‐8)], and humanized chimaeric IgM anti‐Jsb [human immunochemistry monoclonal antibody‐8 (HIMA‐8)] by haemagglutination and gel cards. RBC samples that were DAT positive were used to evaluate the humanized chimaeric IgM monoclonal anti‐Fya (HIMA‐19). RBC samples (n = 243) of known Jsb type were tested in parallel with MIMA‐8 and reagent anti‐Jsb, and 132 samples were tested with MIMA‐8 in gel cards and HIMA‐8 by direct tube testing. No discrepant results were obtained. DAT‐positive RBC samples (n = 27) were correctly phenotyped using HIMA‐19. We conclude that MIMA‐8 is suitable for screening donors and typing patient RBCs. Testing MIMA‐8 with gel cards containing anti‐mouse IgG enables the screening of donors by automated methods. Humanized chimaeric IgM anti‐Jsb and anti‐Fya are suitable as typing reagents by direct agglutination methods.
Synthesis of osteoprotegerin and RANKL by megakaryocytes is modulated by oestrogen
Authors: Bord S. , Frith E. , Ireland D. C. , Scott M. A. , Craig J. I. O. , Compston J. E.
Published: July 2004

To investigate the mechanisms by which megakaryocytes (MKs) may influence bone remodelling, CD34+ cells were cultured for 6, 9 and 12 d with or without 17β‐oestradiol (E) and immunolocalized for osteoprotegerin (OPG), receptor activator of nuclear factor (NF)‐κB ligand (RANKL) and CD61. Specific protein expression was measured quantitatively by image analysis. Fluorescence‐based immunocytochemistry was used to co‐localize OPG and RANKL with CD61. OPG and RANKL mRNA was assessed in CD61+ cells with or without E at 24 and 48 h. At 6 d, OPG and RANKL expression was unchanged by E treatment. At 9 d, the E‐treated cultures with maturing MKs showed a 1·72‐fold (P < 0·01) increase in OPG expression and a 1·8‐fold (P < 0·01) reduction in RANKL. Maximal OPG expression was seen at 12 d with a threefold induction of expression (P < 0·001), whilst RANKL levels were further suppressed by 2·3‐fold compared with controls (P < 0·001). CD61 co‐localized with OPG and RANKL. mRNA data were consistent with that of protein, with a 90‐fold induction in OPG expression and a 34‐fold suppression of RANKL expression by E (P < 0·001). Thus, E stimulates megakaryocytopoiesis and modulates OPG and RANKL expression, providing evidence that MKs may play a role in bone remodelling and, in particular, in E‐induced changes in osteoclastogenesis and bone resorption.
Neutrophil CD177 (NB1 gp, HNA‐2a) expression is increased in severe bacterial infections and polycythaemia vera
Authors: Göhring K. , Wolff J. , Doppl W. , Schmidt K. L. , Fenchel K. , Pralle H. , Sibelius U. , Bux J.
Published: July 2004

The NB1 glycoprotein (CD177, HNA‐2a antigen) is exclusively expressed on human neutrophils. As the clinical significance of CD177 expression is unknown, we investigated its expression in healthy individuals before and after stimulation with granulocyte colony‐stimulating factor (G‐CSF), in patients with rheumatoid arthritis, viral hepatitis, severe bacterial infections and polycythaemia vera. Expression was quantitatively determined by flow cytometry and by real time polymerase chain reaction. Only G‐CSF‐stimulated individuals and patients with severe bacterial infections and polycythaemia showed a significantly (P < 0·001) increased CD177 expression compared with healthy individuals, indicating that neutrophil CD177 expression can increase significantly in certain clinical conditions.
6,7‐Dihydroxy‐3‐phenylcoumarin inhibits thromboplastin induced disseminated intravascular coagulation
Authors: Tunali Tugba , Yarat Aysen , Bulut Mustafa , Emekli Nesrin
Published: July 2004

6,7‐Dihydroxy‐3‐phenylcoumarin (DHPC) was tested to determine whether it had any effect on vitamin K inhibition, by investigating the prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen level and platelet count. The anticoagulant and antithrombotic effects of DHPC were compared with those of warfarin by conducting a 4 h acute trial on thromboplastin‐induced disseminated intravascular coagulation (DIC), investigating various haemostatic and antioxidant system parameters and performing a haemogram. Of most significance was that in the 5‐d DHPC trial on healthy controls, PT, APTT, fibrinogen, platelet count remained within normal levels. In the 4‐h DIC trial, both DHPC (0·025 mg/kg, i.v.) and warfarin (0·25 mg/kg, i.v.) significantly inhibited DIC, by reducing the PT, APTT, and fibrin degradation products and increasing fibrinogen levels and platelet count. In the DIC drug groups, lipid peroxidation significantly increased only in the warfarin group and glutathione significantly increased only in the DHPC group. However leucocyte count was significantly higher in the DHPC than the warfarin group. Further investigation is required for why DHPC is effective on the parameters investigated, at doses one‐tenth of those of warfarin.
Dehydrated hereditary stomatocytosis is associated with neonatal hepatitis
Authors: Rees David C. , Portmann Bernard , Ball Colin , Mieli‐Vergani Giorgina , Nicolaou Anna , Chetty Margaret C. , Stewart Gordon W.
Published: July 2004

Dehydrated hereditary stomatocytosis (DHSt) is an inherited haemolytic anaemia associated with increased red cell membrane permeability to Na+ and K+. It is increasingly recognized that a syndrome of self‐limiting perinatal ascites can accompany the haemolysis. The cause of the perinatal ascites is unknown, and it has been argued that this could be due to cardiovascular, hepatic or lymphatic problems. We describe the case of a 16‐year‐old girl who presented neonatally with abnormal liver function tests and ascites. She was extensively investigated at that time. A liver biopsy showed hepatitis and fatty changes. Her ascites resolved within 6 months. At the age of 15 years, she developed an episode of acute haemolysis and was re‐investigated. A diagnosis of DHSt was made. Pseudohyperkalaemia, due to ex vivo loss of K+ from red cells, was present. This study confirms the previously noted association of DHSt, pseudohyperkalaemia and perinatal ascites, and suggests that the latter is of predominantly hepatic origin.
Predictors of behavioural and emotional problems of children placed in children's homes in Croatia
Authors: Franz Branka Sladovic
Published: July 2004

Since children's homes in Croatia and elsewhere nowadays deal with children who have more problems than in the past, there is a need to identify predictors of the children's behavioural and emotional problems in order to provide adequate interventions and an appropriate developmental environment. This research was carried out in all children's homes in Croatia. The sample consisted of 263 children aged 10–14. Standardized instruments (Youth Self‐Report and Child Behaviour Check‐List) were used as outcome measures of children's psychosocial functioning. The results showed significantly more behavioural and emotional problems in children in children's homes than in the comparison group of children living with their families. Child care‐workers assessed fewer behavioural and emotional problems than did the children themselves. Multiple regression analyses were used to determine predictors for well‐being scores. The level of everyday stress in children's homes, coping strategies, perception of social support and school achievements proved to be the most significant predictors. Since those variables belong to mediating variables within the suggested model of research, it is argued that positive changes in these areas can facilitate a better psychosocial functioning of the children in children's homes.
Cognitive‐behavioural interventions for adolescents in residential child care in Scotland: an examination of practice and lessons from research
Authors: Stevens Irene
Published: July 2004

This paper sets out to examine the basis and use of cognitive‐behavioural interventions with adolescents in residential child care. The paper outlines the results of a survey of the use of cognitive‐behavioural interventions in Scotland. The survey indicates that such interventions are used widely in residential schools and secure units in Scotland. The paper then reviews some of the studies relating to cognitive‐behavioural interventions, which appear to be most relevant to residential child care. The review revealed many of the positive outcomes of cognitive‐behavioural interventions. However, there are some cautionary notes highlighted by the survey and the review. These relate to issues about generalization of learning and the meaning of the intervention for the young person and for the staff. The paper discusses the importance of other factors in determining the success of cognitive‐behavioural interventions. These factors include the importance of accurate assessment, the role of staff training and the need to ensure that interventions are always in the best interests of the child.
‘Given up hope of dying’: A child protection approach to deliberate self‐harm in adolescents admitted to a paediatric ward
Authors: Souter Annie , Kraemer Sebastian
Published: July 2004

This paper describes an approach developed by a local authority social worker based in a paediatric department, working in close collaboration with consultant child and adolescent psychiatrists and paediatricians. The literature is informative on the significant psychosocial difficulties of young people who harm themselves deliberately, but there is little published work on the complexities of the actual multidisciplinary care given to these patients, the majority of whom are not actively suicidal by the time they are interviewed. The task is as much to intervene in the lives of young people at risk of harm or neglect as to prevent suicide.
Type 3 von Willebrand disease: assessment of complications and approaches to treatment – results of a patient and Hemophilia Treatment Center Survey in the United States
Authors: Sumner M. , Williams J.
Published: July 2004

Summary.  It has been reported many times that the optimal treatment of patients with von Willebrand disease (VWD) has yet to be determined. To assist in achieving this goal, the aim of this survey was as follows: to collect information on current treatment practices; to determine the impact of the disease in type 3 VWD patients in the USA and to gain insight into the perceived and actual role that prophylaxis therapy plays in patient management. Nurse coordinators of the 146 federally funded Hemophilia Treatment Centers (HTCs) in the USA were contacted and informed of the survey and its goals. Those caring for type 3 VWD patients and willing to participate in the survey were mailed two questionnaires. The first questionnaire addressed the overall VWD patient population at their centre and asked questions relating to treatment practices. The second questionnaire focused on the impact that type 3 VWD has upon a patient's life. Thirty‐four centres responded to the survey, providing a 48% response rate within the available pool of those centres treating type 3 VWD patients. Seventy‐nine patients, with a mean age of 24.2 years, reported their primary causes of bleeding as; epistaxis (53.9%); joint bleeds (40.9%); dental bleeds (27.6%) and heavy menstrual bleeding (23.7%). There were a mean number of 9.2 reported non‐life‐threatening bleeds per year and a mean of 0.7 reported life‐threatening bleeds per year. Forty percent reported lost days from school or work, caused by bleeds or other VWD complications. Sixty‐two percent reported no joint damage. Regarding current treatment practices, 88.6% of respondents stated that their centre treated on demand and 13.9% used a prophylactic treatment regimen. Seventy‐three percent of respondents felt that prophylaxis was either very or extremely helpful. This survey helps to document current treatment practices and morbidity exhibited by type 3 VWD patients. The results highlight the discrepancy between the use of prophylactic therapy, its perceived value and the level of joint damage and other morbidity experienced in this population.
Haemophilia and thrombophilia: an unexpected association!
Authors: Dargaud Y. , Meunier S. , Negrier C.
Published: July 2004

Summary.  In patients with haemophilia, a close correlation is usually observed between the clinical expression of the disease and plasmatic factor VIII/factor IX clotting activity. However, some patients experience milder bleeding phenotypes than others, although they exhibit a similar biological profile. The high prevalence of some inherited thrombophilia risk factors offers the possibility of a co‐inheritance in haemophilic patients which could influence the phenotypic expression of the disease. Rare thrombotic complications occurring in haemophiliacs could also be facilitated by the co‐inheritance of modifier genes. The majority of thrombotic events occurring in haemophiliacs are in relation to clotting factor infusions or central venous catheters. Concerning surgical situations, in the absence of therapeutic recommendations, postoperative thromboprophylaxis is not systematically performed in haemophiliacs. However, substitutive treatment more or less completely corrects the coagulation defect and makes the venous thrombosis risk closer to the control population. It should be emphasized that haemophilia does not fully protect against venous thromboembolic disease. Patients with haemophilia very infrequently experience thrombotic events. Thus, the management of thrombotic complications occurring in haemophilic patients should be discussed in each case according to the precipitating risk factors, the clinical context and the thrombo‐haemorrhagic balance of the patient with respect to a particular clinical situation.
Safety and supply of haemophilia products: worldwide perspectives
Authors: Farrugia A.
Published: July 2004

Summary.  The survival and well‐being of people with haemophilia depends on the supply of safe therapeutic products. Safety and supply are entirely intertwined principles; in the absence of adequate amounts of coagulation products, safety measures may be compromised in order to enhance supply, leading to risks which may result in morbidity and mortality. As haemophilia therapy has emerged through the development of blood transfusion and plasma fractionation, the safety of the blood supply in general has had a strong effect on haemophilia care. Despite the gradual detachment of haemophilia care from blood transfusion through the use of recombinant products, the majority of the world's population with haemophilia in the developing world will be reliant on blood products for the foreseeable future. It is, therefore, important to continue efforts for a safe and sufficient blood supply worldwide. As such a blood supply develops, possibly in tandem with an independent plasma fractionation industry, the level of haemophilia care should improve with the gradual introduction of concentrates for the ultimate goal of covering all aspects of care. Constant vigilance for the threat of blood‐borne pathogens should be linked to considerations of how these products are to be manufactured. This should be governed entirely by considerations of safety and pharmaceutical competence. Of equal importance is a governmental capacity to oversee the entry and maintenance of these products on the market. While it is not possible for all countries to have a regulatory authority of the same status as that of the developed countries, it is perfectly feasible to develop a set of basic principles which allow an assessment of basic product safety, quality and efficacy to be made.
Neurosurgical intervention in a haemophilic child with a subdural and intracerebral haematoma
Authors: Cermelj M. , Negro F. , Schijman E. , Ferro A. M. , Acerenza M. , Pollola J.
Published: July 2004

Summary.  A case of a 3‐year‐old boy with severe haemophilia A who had a successful neurosurgical drainage of a combined spontaneous left temporal subdural and intra‐parenchimal haematoma is reported. Surgical intervention was required because of clinical worsening during conservative treatment with dexamethasone and factor VIII (FVIII) replacement therapy. Continuous FVIII infusion was given before, during and after the procedure. There were no surgical complications and neurological examination remains intact. Neurosurgical interventions may be reserved for special, high‐risk cases, as the one presented.
Development of a new disease‐specific quality‐of‐life questionnaire to adults living with haemophilia
Authors: Arranz P. , Remor E. , Quintana M. , Villar A. , Díaz J. L. , Moreno M. , Monteagudo J. , Ugarriza A. , Soto I. , Pérez R. , Chacón J. , García‐Luaces M. , Cid A. , Balda I. , López M. F. , Gutíerrez M. J. , Martínez E. , Marrero C. , Prieto M. , Sedano C. , Vaca R. , Altisent C. , Hernández‐Navarro F. ,   Hemofilia‐QoL Group
Published: July 2004

Summary.  A haemophilia‐specific health‐related quality‐of‐life questionnaire (named ‘Hemofilia‐QoL’) was developed to assess quality‐of‐life in adults with haemophilia, and was psychometrically tested. Seventy‐three interviews with haemophilia patients and health care professionals were used to generate the items included in the questionnaire, and expert ratings on the items formulated were used to screen them for potential omission. This was followed by psychometric testing in a sample of 35 patients. Preliminary psychometric testing of the revised questionnaire version, which contains 10 domains (physical health, physical role, joint damage, pain, treatment satisfaction, emotional role, mental health, social support), showed acceptable reliability (α = 0.94 for the Hemofilia‐QoL total score) and validity, and this will be examined in a subsequent study with a larger patient sample.
Unresponsiveness to factor VIII inhibitor bypassing agents during haemostatic treatment for life‐threatening massive bleeding in a patient with haemophilia A and a high responding inhibitor
Authors: Hayashi T. , Tanaka I. , Shima M. , Yoshida K. , Fukuda K. , Sakurai Y. , Matsumoto T. , Giddings J. C. , Yoshioka A.
Published: July 2004

Summary.  We report a case of haemophilia A with a high responding inhibitor of factor VIII (FVIII) who had a serious retroperitoneal haematoma caused by penetration of a duodenal ulcer. Inhibitor‐bypassing therapy was commenced immediately on admission. On the 17th day of treatment with activated prothrombin complex concentrate (APCC; FEIBA®), re‐bleeding occurred and thrombelastography (TEG) demonstrated resistance to therapy. Treatment was changed to recombinant activated factor VII (rFVIIa; NovoSeven®) and resulted in clinical improvement together with an improvement in TEG parameters. On the 10th day of continuous infusion with NovoSeven®, however, TEG again showed resistance to therapy. FEIBA® infusions were re‐introduced and TEG results remained satisfactory for 7 days. On day 34, however, further retroperitoneal bleeding was evident and a decline in the haemostatic efficiency of FEIBA® was recorded by TEG. NovoSeven® was again successfully administered for 7 days. There were no laboratory findings to indicate disseminated intravascular coagulation (DIC), hypercoagulability or abnormal fibrinolysis. The plasma‐based clotting tests did not show any additional prolongation on the occasions when the TEG demonstrated unresponsiveness to FEIBA® or NovoSeven®. These findings suggested that some component of whole blood, other than plasma might have governed the TEG data. The long‐term use of APCC such as FEIBA® or rFVIIa, requires careful monitoring in terms of FVIII inhibitor bypassing activity as well as the tendency to DIC.
Pharmacokinetics of activated recombinant coagulation factor VII (NovoSeven®) in children vs. adults with haemophilia A
Authors: Villar A. , Aronis S. , Morfini M. , Santagostino E. , Auerswald G. , Thomsen H. F. , Erhardtsen E. , Giangrande P. L. F.
Published: July 2004

Summary.  To establish the pharmacokinetic profile of activated recombinant coagulation factor VII (rFVIIa; NovoSeven®) in children with haemophilia A, and to compare it with the pharmacokinetic profile in adults with haemophilia A.

Twelve children (2–12 years) received one single dose of rFVIIa 90 and 180 μg kg−1 in randomized order separated by a washout period of 48 h to 1 month. Six adults (18–55 years) received a single dose of rFVIIa 90 μg kg−1. The pharmacokinetic analyses were based on a non‐compartmental method.

In children, the plasma level of FVII increased proportionally with the dose. The total body clearance normalized for body weight was significantly faster in children than in adults (FVII:C, 58 vs. 39 mL kg−1 h−1 and FVIIa, 78 vs. 53 mL kg−1 h−1, P < 0.05). A trend towards a larger volume of distribution at steady‐state in children than in adults was observed (P > 0.05).

Dose proportionality was established for plasma concentrations of FVII in children with haemophilia A at the dose levels investigated (90 and 180 μg kg−1 rFVIIa). Following administration of rFVIIa 90 μg kg−1, significantly faster clearance was observed in children compared with adults, suggesting that higher doses of rFVIIa may be needed to achieve the same plasma levels as in adults.
Prospects for gene therapy of haemophilia
Authors: Nathwani A. C. , Davidoff A. M. , Tuddenham E. G. D.
Published: July 2004

Summary.  That gene therapy offers the promise of a cure for haemophilia was apparent more than a decade ago. After years of failure, substantial progress in the efficiency of gene transfer technology has recently resulted in impressive success in animal models with haemophilia. However, fears of the risks intrinsic to such therapy have been raised by the fate of two children cured of immune deficiency by gene transfer who have, however, subsequently developed leukaemia as a result of insertional mutagenesis. The purpose of this review is to outline the current status of gene therapy in light of recent successes and tragedies and to consider the prospects for curing haemophilia in the short‐to‐medium term.
Comparison of attitudes towards prenatal diagnosis and termination of pregnancy for haemophilia in Iran and Italy
Authors: Karimi M. , Peyvandi F. , Siboni S. , Ardeshiri R. , Gringeri A. , Mannucci P. M.
Published: July 2004

Summary.  Prenatal diagnosis (PND) is an important issue in the comprehensive care of haemophiliacs. As a consequence of technological progress made in the field of PND, the early detection of an affected fetus provides the expectant couple with a chance to terminate pregnancy. This study was undertaken to assess the attitudes of two different haemophilic populations in Iran and Italy towards PND and termination of pregnancy. This study series included 59 Iranians (38 haemophilia A patients and 21 mothers) and 50 Italians (27 haemophilia A patients, 16 mothers and seven fathers). All the 109 participants received a questionnaire including demographic characteristics and evaluating the psychological effects stemming from PND and termination of pregnancy. Approximately 84.7% of the Iranians and 35.4% of the Italians were not familiar with the possibilities afforded by PND for haemophilia (P < 0.001). Termination of pregnancy appeared to be accepted by 58.2% of the Iranian and 16.7% of the Italian participants (P < 0.001). The greater rate of acceptability of abortion in Iranians may be due to differences in the quality of patient care in the two countries.
Cytomegalovirus infection in symptomatic periapical pathosis
Authors: Slots J. , Nowzari H. , Sabeti M.
Published: July 2004

Aim  To compare the presence of human cytomegalovirus (HCMV) and Epstein–Barr virus (EBV) infections in samples from 25 symptomatic and 19 asymptomatic periapical lesions.

Methodology  Periapical samples were collected by sterile curettes in conjunction with apicectomy. cDNA‐based HCMV and EBV identification was performed on total mRNAs extracted from peripapical tissues, using primers for genes transcribed during the productive phase of the herpesvirus infection. Statistical analysis was performed using chi‐squared test.

Results  HCMV was detected in 100% of the symptomatic and in 37% of the asymptomatic study lesions. EBV was identified only in HCMV‐infected periapical lesions. The difference in occurrence of HCMV and EBV between symptomatic and asymptomatic periapical lesions was statistically significant (P < 0.0001).

Conclusions  The noteworthy finding of this study was the ubiquitous occurrence of HCMV active infection in symptomatic periapical pathosis. EBV may contribute to periapical pathogenesis in a subset of symptomatic lesions. HCMV and EBV infections may cause periapical pathosis by inducing cytokine and chemokine release from inflammatory or connective tissue cells, or by impairing local host defences resulting in heightened virulence of resident bacterial pathogens. Knowledge about the role of herpesviruses in periapical pathosis seems important to fully delineate the pathogenesis of endodontic infectious diseases. HCMV and probably EBV should be added to the list of putative pathogenic agents in symptomatic periapical disease.
The efficacy of pain control following nonsurgical root canal treatment using ibuprofen or a combination of ibuprofen and acetaminophen in a randomized, double‐blind, placebo‐controlled study
Authors: Menhinick K. A. , Gutmann J. L. , Regan J. D. , Taylor S. E. , Buschang P. H.
Published: July 2004

Aim  To compare ibuprofen, to an ibuprofen/acetaminophen combination in managing postoperative pain following root canal treatment. It is hypothesized that the drug combination will provide more postoperative pain relief than the placebo or ibuprofen alone.

Methodology  Patients presenting at the Texas A&M Baylor College of Dentistry's graduate endodontic clinic, experiencing moderate to severe pain, were considered potential candidates. Fifty‐seven patients were included based on established criteria. Following administration of local anaesthesia, a pulpectomy was performed. The patients were administered a single dose of either: (i) placebo; (ii) 600 mg ibuprofen; or (iii) 600 mg ibuprofen and 1000 mg of acetaminophen. Patients recorded pain intensity following treatment on a visual analogue scale and a baseline four‐point category pain scale as well as pain relief every hour for the first 4 h then every 2 h thereafter for a total of 8 h. A general linear model (GLM) analysis was used to analyse the outcome.

Results  Based upon the GLM analysis, there was a significant difference between the ibuprofen and the combination drug group, and between placebo and combination drug groups. There was no significant difference between the placebo and the ibuprofen.

Conclusion  The results demonstrate that the combination of ibuprofen with acetaminophen may be more effective than ibuprofen alone for the management of postoperative endodontic pain.
A clinical report of 85 fractured metallic post‐retained crowns
Authors: Fox K. , Wood D. J. , Youngson C. C.
Published: July 2004

Aim  To determine any patient, technique or material factors that were significantly associated with post fracture in metallic post systems.

Summary  Eighty‐five fractured metal posts were retrieved over a 5‐year period from a total of 84 patients who attended a dental hospital for treatment. A record was kept of the patient's dentition to identify the tooth with the fractured post, the number of teeth present, the excursive guidance scheme and the likely post design and material. Radiographs of 67 of these teeth were available. The presence or absence of a periapical lesion was noted as was the quality of the root filling. Maxillary lateral incisors, followed by maxillary centrals were associated with most of the fractured posts. The median survival to fracture was 36 months. The number of teeth in the dentition had a mode of 28. The most commonly fractured post design was a serrated and parallel design with an average diameter of 1.2 mm at the site of fracture. Apical lesions were found in 67% of the teeth with radiographs. Only 12% of teeth with a ‘good’ apical filling as opposed to 55% with a ‘poor’ apical filling had apical lesions.

Key learning points

Maxillary lateral incisors, followed by maxillary central incisors are at greatest risk of having a fractured metallic post.

Having a large number of teeth in the dentition or an adequate length of post was not protective against metallic post fracture.

Teeth with fractured post‐retained crowns had a high incidence of apical lesions.
Assessment of infectious salmon anaemia virus prevalence for different groups of farmed Atlantic salmon, Salmo salar L., in New Brunswick
Authors: McClure C A , Hammell K L , Dohoo I R , Nerette P , Hawkins L J
Published: July 2004

Infectious salmon anaemia (ISA) virus (ISAV) has been causing disease in New Brunswick since 1996. As a control measure, all fish in an outbreak cage are killed. The objective of this study was to compare ISAV prevalence in cages experiencing an outbreak with healthy cages from the same farm, neighbouring farms and distant farms. Atlantic salmon from five different groups were tested using an RT‐PCR test. Groups included moribund fish from a cage experiencing an outbreak (A), healthy fish from an outbreak cage (B), healthy fish from a negative cage from a farm experiencing an outbreak in a different cage (C), healthy fish from a negative farm near an outbreak farm (D), and healthy fish sampled at a negative farm located in an area with only negative farms (E). Apparent prevalences (standard error) for the different groups (A–E) were 0.94 (±0.026), 0.41 (±0.062), 0.29 (±0.040), 0.08 (±0.037) and 0.08 (±0.037), respectively. All groups were significantly different (P < 0.002) from each other except for groups B and C and groups D and E. Because the prevalence of the virus was significantly higher in the outbreak cage (B) compared with other sites, early harvest of outbreak cages will remove one source of virus. However, ISA negative cages (C) that remain on the positive farm may potentially act as a viral reservoir.
Investigation of wild caught whitefish, Coregonus lavaretus (L.), for infection with viral haemorrhagic septicaemia virus (VHSV) and experimental challenge of whitefish with VHSV
Authors: Skall H F , Kjær T E , Olesen N J
Published: July 2004

One hundred and forty‐eight wild whitefish, Coregonus lavaretus (L.), were caught by electrofishing and sampled for virological examination in December 1999 and 2000, during migration from the brackish water feeding grounds to the freshwater spawning grounds, where the whitefish may come into contact with farmed rainbow trout. All samples were examined on cell cultures. No viruses were isolated. Three viral haemorrhagic septicaemia virus (VHSV) isolates of different origin were tested in infection trials by immersion and intraperitoneal (IP) injection, using 1.5 g farmed whitefish: an isolate from wild caught marine fish, a farmed rainbow trout isolate with a suspected marine origin and a classical freshwater isolate. The isolates were highly pathogenic by IP injection where 99–100% of the whitefish died. Using an immersion challenge the rainbow trout isolates were moderately pathogenic with approximately 20% mortality, whereas the marine isolate was virtually non‐pathogenic. At the end of the experiment it was possible to isolate VHSV from survivors infected with the marine and suspected marine isolates. Because of the low infection rate in wild whitefish in Denmark, the role of whitefish in the spread of VHSV in Denmark is probably not significant. The experimental studies, however, showed that whitefish are potential carriers of VHSV as they suffer only low mortality after infection but continue to carry virus.
Histopathological studies on viral nervous necrosis of sevenband grouper, Epinephelus septemfasciatus Thunberg, at the grow‐out stage
Authors: Tanaka S , Takagi M , Miyazaki T
Published: July 2004

Viral nervous necrosis caused by sevenband grouper nervous necrosis virus (SGNNV) has occurred in grow‐out stages (0–3 years old) of sevenband grouper, Epinephelus septemfasciatus, since the 1980s. In the present study, based on histopathological features of the central nervous system (CNS) in naturally diseased fish, pernasal infection experiments using grow‐out fish were performed and pernasal infection was established as a putative invasion route of SGNNV. The definite SGNNV‐targeted cells were determined by histopathological studies including indirect fluorescent antibody test and electron microscopy. Nerve cells in the olfactory lobe were most extensively necrotized with vacuolation followed by infiltration of microglia and macrophages. Purkinje cells and Golgi cells were extensively infected in the cerebellum. Megalocells and small nerve cell nuclei were also infected in the preoptic area, thalamus, medulla oblongata and spinal cord. Only a few small nerve cells were infected in the olfactory bulb and optic tectum. The retina of some diseased fish displayed vacuolated bipolar cells of the inner nuclear layer and in the ganglion cell layer. These SGNNV‐infected nerve cells displayed viroplasmic inclusions containing virions, vacuoles and myelin‐like structures. Based on observed histopathological changes, the lesion of the CNS was characterized by encephalitis but not encephalopathy.
Lymphoproliferative disorders in chronic hepatitis C
Authors: Idilman R. , Colantoni A. , De Maria N. , Alkan S. , Nand S. , Van Thiel D. H.
Published: July 2004

Summary.  Chronic hepatitis C virus (HCV) infection is associated with the development of lymphoproliferative disorders (LPDs). The aim of this investigation was to determine the prevalence and characterization of monoclonal gammopathy and benign and malignant LPDs in individuals with chronic hepatitis C. A total of 233 subjects diagnosed with chronic hepatitis C (male/female ratio: 131/102, median age; 49 years) were studied. Serum and urine were examined for the presence of a monoclonal gammopathy. A bone marrow aspirate and biopsy was obtained in individuals with a monoclonal gammopathy. Thirty‐two patients (13.7%, 32 of 233) had a monoclonal gammopathy; 75% of them were benign and were not associated with malignant disorders (24 of 32) while 25% were associated with malignant LPDs or a plasma cell disorder (eight of 32). Two additional subjects without monoclonal gammopathy were diagnosed as having a malignant LPDs. The prevalence of malignant LPDs/plasma cell disorder in individuals with HCV‐induced chronic liver disease was 4.3%. No difference was found in terms of disease duration, HCV genotype, viral load, alanine aminotransferase level or histopathologic score between the subjects with or without a monoclonal gammopathy. The presence of mixed cryoglobulinaemia was strongly associated with the presence of an underlying malignant disorder. Hence a monoclonal gammopathy is found in 14% of patients with chronic hepatitis C and is associated with malignant B‐cell LPD in more than a quarter of such patients. The prevalence of LPDs in individuals with HCV‐induced chronic liver disease is greater than that of the normal healthy population.
Mutations in the putative HCV‐E2 CD81 binding regions and correlation with cell surface CD81 expression
Authors: Kronenberger B. , Sarrazin C. , Hofmann W. P. , Von Wagner M. , Herrmann E. , Welsch C. , Elez R. , Rüster B. , Piiper A. , Zeuzem S.
Published: July 2004

Summary.  The hepatitis C virus (HCV) envelope (E)2 protein interacts with the cellular receptor CD81 leading to modulation of B and T cell function. Recently, a higher binding affinity of subtype 1a in comparison with 1b derived E2 proteins for CD81 in vitro was described. The importance of mutations within the putative CD81 binding regions of different HCV geno‐/subtypes in correlation with CD81 expression is unknown. In the present study, CD81 expression on blood lymphocytes of patients with chronic hepatitis C infected with different HCV geno‐/subtypes were analysed by fluorescence activated cell sorter analyses. In addition, the putative CD81 binding regions on the E2 gene comprising the hypervariable region (HVR)2 were analysed by direct sequencing. CD81 expression on CD8(+) T‐lymphocytes from patients infected with subtype 1a (n = 6) was significantly higher in comparison with subtype 1b (n = 12) and 3 (n = 5) infected patients before and during antiviral therapy (P = 0.006; P = 0.021, respectively). Sequencing of the putative CD81 binding regions in the E2 protein comprising the HVR2 (codon 474–495 and 522–552 according to the HCV‐1a prototype HCV‐H) showed a highly conserved motif within HVR2 for subtype 1a isolates and an overall low number of mutations within the putative CD81 binding regions, whereas numerous mutations were detected for subtype 1b isolates (12.0 vs 23.6%). HCV‐3 isolates showed an intermediate number of mutations within the putative binding sites (19.2%; P = 0.022). In conclusion, the highly conserved sequence within HVR2 and putative CD81 binding sites of subtype 1a isolates previously associated with a high CD81 binding affinity in vitro is correlated with high CD81 expression on CD8(+) T‐lymphocytes in vivo.
Analysis of CCR5‐Δ32 and CCR2‐V64I polymorphisms in a cohort of Spanish HCV patients using Real‐time Polymerase Chain Reaction and Fluorescence Resonance Energy Transfer technologies
Authors: Ruiz‐Ferrer M. , Barroso N. , Antiñolo G. , Aguilar‐Reina J.
Published: July 2004

Summary.  Nowadays it is clear that chemokine–chemokine receptor interactions are important in chronic hepatitis C virus (HCV) infection. The objective of the present study was to elucidate the involvement of the CCR5‐Δ32 and CCR2‐V64I polymorphisms in the response to the HCV infection, as well as in the histological damage and the outcome of the infection. A cohort of 139 patients with hepatitis C and 100 healthy blood donors were analysed for both polymorphisms using real‐time polymerase chain reaction (PCR) and LightCyclerTM technology. We have detected the CCR5‐Δ32 allele in 15 of 278 HCV chromosomes (5.4%) and 15 of 200 control chromosomes (7.5%). The CCR2‐V64I allele was present in 24 of 278 HCV chromosomes (8.6%) and 19 of 200 control chromosomes (9.5%). Analysis of the histological parameters showed no statistical significance when comparing the patients carrying the variants vs the cases with the wild‐type allele. Our results seem to indicate that the CCR5‐Δ32 and CCR2‐V64I polymorphisms are not related to the response to HCV infection, histological damage and outcome of infection in our cohort of Spanish HCV patients.
Occult hepatitis B virus infection in Greek patients with chronic hepatitis C and in patients with diverse nonviral hepatic diseases*
Authors: Georgiadou S. P. , Zachou K. , Rigopoulou E. , Liaskos C. , Mina P. , Gerovasilis F. , Makri E. , Dalekos G. N.
Published: July 2004

Summary.  Occult hepatitis B virus (HBV) infection has been reported in patients with chronic hepatitis C who are negative for HBV surface antigen (HBsAg). However, the significance of ‘silent’ HBV in hepatitis C virus (HCV) infection is unknown. We investigated 540 subjects for the presence of occult HBV in Greek HCV patients, patients with nonviral liver diseases and healthy donors in an attempt to determine the frequency and importance of this phenomenon. One hundred and eighty‐seven anti‐HCV(+)/HBsAg(−) patients’ sera were investigated for the presence of HBV‐DNA by polymerase chain reaction. Two hundred and eighty‐two selected blood donors (positive for antibodies to HBV core antigen) and 71 patients with various nonviral hepatic diseases consisted the control groups [both controls were anti‐HCV(−)/HBsAg(−)]. HBV‐DNA was detected in 26.2% of HCV‐infected patients vs 8.5% of patients with nonviral diseases (P = 0.003) and 0/282 of donors (P = 0.0000). HBV‐DNA was neither associated with HBV markers, nor with the clinical status of HCV and nonHCV patients. Neither epidemiological, histologic and virologic data nor the response to therapy were associated with the HBV‐DNA detection. Hence one quarter of HCV‐infected patients had occult HBV infection. Similar findings were not found in both control groups. Occult HBV infection in Greek patients with chronic hepatitis C does not seem to modify the progression of chronic liver disease. Further studies of longer duration are needed in order to clarify the role of ‘silent’ HBV infection in HCV‐infected patients.
The activity of a single‐stranded promoter of plasmid ColIb‐P9 depends on its secondary structure
Authors: Nasim M. T. , Eperon I. C. , Wilkins B. M. , Brammar W. J.
Published: July 2004

The leading region of the conjugal bacterial plasmid ColIb‐P9 contains three dispersed repeats of a 328 bp sequence homologous to Frpo, a sequence from plasmid F that acts as a promoter in single‐stranded DNA. One of these sequences, ssi3, inactive in the double‐stranded form, promoted in vitro transcription exclusively from the single strand that is transferred during conjugation. Promoter activity was dependent on the presence of RNA polymerase holoenzyme containing sigma 70. Transcription initiated from the position predicted from folding the single‐stranded DNA to form a pseudo double‐stranded hairpin structure containing recognizable −35 and −10 promoter elements. Footprinting of RNA polymerase holoenzyme on single‐stranded ssi3 DNA was consistent with this suggestion. Mutagenesis of the putative −35 region inactivated the promoter, but random mutations in the −10 region had little effect. The putative −10 region is a poor match to the consensus sequence and contains mismatched bases. Elimination of these mismatches invariably destroyed single‐strand promoter activity. These observations reveal the crucial contribution of the unpaired bases in the −10 region in potentiating the formation of the productive open complex with RNA polymerase.
Use of proteomics to identify novel virulence determinants that are required for Edwardsiella tarda pathogenesis
Authors: Srinivasa Rao P. S. , Yamada Yoshiyuki , Tan Yuen Peng , Leung Ka Yin
Published: July 2004

Edwardsiella tarda is an important cause of haemorrhagic septicaemia in fish and also of gastro‐ and extraintestinal infections in humans. Using a combination of comparative proteomics and TnphoA mutagenesis, we have identified five proteins that may contribute to E. tarda PPD130/91 pathogenesis. Lowered protein secretion, impaired autoaggregation and the absence of six proteins were observed only in three highly attenuated mutants when cultured in Dulbecco's modified eagle medium (DMEM). Five out of six proteins could be identified by their mass spectra. Three proteins were identified as putative effector proteins (EseB, EseC and EseD) that are homologous to SseB, SseC and SseD of a type III secretion system (TTSS) in Salmonella species. The other two were EvpA and EvpC, homologous to Eip20 and Eip18 in Edwardsiella ictaluri. The complete sequencing and homology studies of evpA–H indicate that similar gene clusters are widely distributed in other pathogens such as Escherichia, Salmonella, Vibrio and Yersinia species with unknown functions. Insertional inactivation and deletion of evpB or evpC led to lower replication rates in gourami phagocytes, and reduced protein secretion and virulence in blue gourami. Complementation of these deletion mutants showed partial recovery in the above three phenotypes, indicating that these genes are vital for E. tarda pathogenesis. The transport of the EvpC protein may not use the TTSS in E. tarda. The expression of EvpA and EvpC as well as EseB, EseC and EseD was temperature dependent (suppressed at 37°C), and disruption of esrB affected their expression. The present study identifies two possible secretion systems (TTSS and Evp) that are vital for E. tarda pathogenesis.