bluuebunny/crossref_metadata_2025 huggingface dataset:
Utilisation of Domiciliary Dental Services
Authors: Kinsey J G , Winstanley R B
Published: January 2004

Objectives: To develop undergraduate dental student understanding of the attitudes of elderly people towards dentistry and of the barriers which prevent them from seeking treatment. Design: Each student interviewed a) an elderly person already known to them and b) an elderly person at a Day Rehabilitation Unit using a questionnaire. Students were not trained nor standardised. Setting: a) in the student's home locality, b) in a Rehabilitation Unit in Sheffield. Subjects: 161 people were interviewed with mean ages a) 78 years and b) 85 years. Main outcome measures: Perceptions of treatment need and domiciliary treatment by these elderly people. The results are summarised but not analysed because of the inherent limitations of data derived by unsupervised students. Conclusions: This educational exercise successfully engaged the studentsí minds and, for some, generated enthusiasm. They achieved valuable insight into the topic and realised that many people who inevitably have dental problems either feel that this is acceptable or do not know that they can obtain domiciliary dental care.
Socio‐medical condition and oral functional status in an older institutionalised population
Authors: Kossioni A E , Karkazis H C
Published: January 2004

Objectives: To investigate the association between the general medical condition, the socio‐economic status, and some factors related to the functional status of the stomatognathic system. Design: A survey in an elderly population. Subjects: 257 older adults, with a mean age of 83.7 years. Setting: Residential homes for the elderly. Intervention: Examination of the medical records on the overall health and the drugs consumed, a structured interview on the socio‐economic status, the complaints for xerostomia, the subjective chewing difficulties, and a clinical evaluation of the number of natural teeth and the number of posterior occluding pairs of teeth contacts (premolars and molars). Results: Multiple pathology and polypharmacy were recorded. 25% of the residents had no occluding posterior tooth contact (natural or prosthetic) and 62% were edentulous. 43% of the residents reported complaints for xerostomia, and 46% for chewing difficulties when eating specific food types. Xerostomic feelings and chewing problems were not related to age. Chewing difficulties were not related to the number of natural teeth, but to the number of posterior occluding teeth contacts, natural or prosthetic (less than two). From all medical conditions examined, only the psychiatric disorders were significantly related to dental status (p<0.05). Moreover, the number of remaining natural teeth was related to socio‐economic status, while the number of posterior occluding teeth contacts was also inversely related to the duration of institutionalisation (p<0.05). Conclusions: Psychiatric disorders, low socio‐economic status and increased duration of institutionalisation were most closely related to poor dental status. The presence of more than two posterior occluding teeth contacts, natural or prosthetic, benefit the very old patient in terms of reduced subjective chewing difficulty. A formal oral care delivery system for the institutionalised elderly, and particularly for those suffering from psychiatric disorders, is imperative.
Factors which are associated with dental decay in the older individual
Authors: Loesche W.J. , Taylor G.W. , Dominguez L.D , Grossman N.S. , Stoll J.
Published: January 2004

Objectives:To improve reliability of salivary bacterial cultures as a surrogate for plaque levels of cariogenic bacterial species by reporting the salivary CFUs of these organisms as a function of the number of teeth. Design:Cross‐sectional collection of data in a convenience sample of adults over 60 years of age. Setting:Hospital Dental clinic, University bacteriology laboratory. Subjects:523 older dentate subjects, average age 70, including 412 subjects who were in an independent living status and 111 in a dependent‐living situation. Main outcome measures: Subjects were examined for decay and the presence of salivary factors including the levels of S. mutans, lactobacilli, yeast and other bacteria. The salivary levels of the bacteria were adjusted for the number of teeth in the mouth, and the resultant values were entered into multivariable logistic regression models along with clinical and other salivary parameters. Results:Mutans streptococci levels reported as CFUs/ml saliva per tooth were significantly associated with coronal surface decay, and lactobacilli, reported in a similar way, were significantly associated with root surface decay. Salivary levels of yeasts, which had previously been associated with decay in this population, were no longer significant using this construct. Conclusions: This construct of reporting salivary bacteriological data as a function of tooth number and per ml saliva could improve the reliability of bacteriological data obtained in epidemiological studies investigating the role of bacteria in dental decay in the elderly.
The effectiveness of 10% chlorhexidine varnish treatment on dental caries incidence in adults with dry mouth
Authors: Banting David W. , Papas Athena , Clark D. Christopher , Proskin Howard M. , Schultz Miklos , Perry Ross
Published: January 2004

Objectives: This study compared a 10% chlorhexidine varnish treatment with placebo and sham treatments for preventing dental caries in adult patients with xerostomia (dry mouth).

Design: The study was a multicentred, randomized, parallel group, double blind, placebo‐controlled clinical trial.

Setting: All examinations and procedures were performed at Tuft's University, Boston, MA, the University of British Columbia, Vancouver, BC or the University of Western Ontario, London, ON.

Subjects: Subjects were adults with recent or current dental caries experience, high salivary levels of cariogenic microorganisms and low salivary flow rates.

Results: 236 subjects completed at least one post‐treatment examination. There were 697 new carious lesions diagnosed, 446 (64%) located on coronal surfaces and 251 (36%) located on root surfaces. The mean attack rate was 0.23 surfaces/100 surfaces at risk. A treatment difference observed between the Active and Placebo groups was statistically significant for root caries increment (p=.02) and total caries increment (p=.03). A treatment difference observed between the Active and Sham groups was not statistically significant for coronal, root or total caries increment. Analysis of variance of treatment group differences was performed using mutans streptococci counts, salivary flow rates, age, sex, caries prevalence, medications, time to first event and early withdrawal as co‐variables. These factors did not meaningfully alter the findings.

Conclusions: The difference between the 10% chlorhexidine varnish and placebo treatments is considered to be highly clinically significant for root caries increment (41% reduction) and for total caries increment (25% reduction) but only for coronal caries increment (14%).
Oral mucous membrane flora in patients using saliva substitutes
Authors: Johansson G. , Andersson G. , Attstöm R. , Edwardsson S.
Published: January 2004

Salivary substitutes are sometimes valuable for elderly people and radiotherapy patients, and may be used indefinitely. It is possible that this change in the ecology may effect the oral flora.

Objective: To analyse the presence of micro‐organisms on oral mucous membranes during use of saliva substitutes.

Design: Cross‐over single‐blind study.

Setting: Clinic for Maxillofacial Surgery, Malmö University Hospital and Department of Oral Microbiology, Malmö University.

Subjects: 19 patients with low salivary secretion who had been radiated for cancer in the head and neck region. Intervention Two saliva substitutes: linseed extract and a carboxymethyl cellulose preparation (Salinum and MAS‐84) were used for 3 week periods.

Measurements: Microbial samples taken, processed and analysed.

Results: No differences were observed when comparing baseline values with the results after the saliva substitutes and no significant differences between the use of different agents.

Conclusion: The study suggests that use of linseed extract and carboxymethyl cellulose preparation during periods of weeks does not influence flora commonly related to caries, periodontitis or infections in the oral mucous membranes.
The influence of rinsing routines on fluoride retention after toothbrushing
Authors: Sjögren K , Melin N‐H
Published: January 2004

Objective: An important determinant of the anticaries effect of brushing with a fluoride dentifrice is the retention of fluoride in the oral fluids. This study measured the influence of various post‐brushing rinsing routines on fluoride retention.

Setting: The Faculty of Odontology, Göteborg University, Sweden.

Subjects: Twenty‐one healthy dental students, aged 23±2 yr.

Desing: A randomised, cross‐over protocol, comparing various post‐brushing rinsing procedures, with the following variables: volume of rinsing water (5 mL vs. 10×3mL) and duration of rinsing (10 vs. 60 s). Samples of whole saliva, interdental fluid and interdental plaque were analysed for fluoride content.

Results: The duration of rinsing did not influence fluoride retention in whole saliva, interdental fluid or interdental plaque. Rinsing with a higher volume of water substantially decreased fluoride retention in all samples.

Conclusion: Brushing with a fluoride dentifrice should be followed by rinsing procedures which enhance the retention of fluoride. Rinsing with large volumes of water decreases fluoride retention in the oral fluids, and should be discouraged.
Maximum bite force after the replacement of complete dentures
Authors: Müller Frauke , Heath M. Robin , Ott Rudolf
Published: January 2004

Objective: The aim of the present study was to investigate whether the maximum bite force (MBF) can be improved by the replacement of complete dentures for elderly people.

Design: Nine edentulous volunteers, mean age 74.2 (± 5.5) years and average denture experience 19.4 ± 19.5 years (1 to 50 years) had replacement dentures made. After a rehearsal session, MBF was recorded with the old dentures, and with the new dentures immediately at insertion, at 3, 8 days, 2–3 weeks, 1, 2, 3 and 6–10 months post insertion (p‐i.). MBF was recorded with the central bearing point method using a full‐bridge strain gauge load cell. Data were analysed off‐line using the mean of two peak readings per patient per session.

Results: The results indicate that MBF tended to be impaired when replacement dentures were first fitted (n.s.). However, this trend reversed during the first month p‐i. for patients with a “moderate” lower ridge resorption of Atwood grade 3 or 4 (n = 5). Patients with more severe lower ridge resorption Atwood grade 5 or 6 (n = 4) showed a significantly lower MBF over the entire observation period (p0.05) and took longer to regain bite strength. Only patients with moderate bone resorption exceeded their pre‐insertion level of MBF within the observation period of 6–10 months p‐i.

Conclusion: The present pilot study suggests that, at least for elderly patients with severe bone resorption, delayed improvement of MBF should be expected with replacement complete dentures.
When Stepfathers Claim Stepchildren: A Conceptual Analysis
Authors: Marsiglio William
Published: January 2004

Guided by social constructionist and symbolic interactionist perspectives and a grounded theory method, my conceptual analysis explores stepfathers’ experiences with claiming stepchildren as their own. Using in‐depth interviews with a diverse sample of 36 stepfathers, my analysis focuses on paternal claiming as a core category and generates 10 properties relevant to this social psychological phenomenon: timing, degree of deliberativeness, degree of identity conviction, paternal role range, solo‐shared identity, mindfulness, propriety work, naming, seeking public recognition, and biological children as benchmarks. In addition, I consider how five conditions may influence stepfathers’ perceptions of stepchildren. My analysis reveals a number of sensitizing concepts for studying stepfathers’ efforts to develop a sense of group belonging and a fatherlike identity.
Role of the Father‐Adolescent Relationship in Shaping Adolescents’ Attitudes Toward Divorce
Authors: Risch Sharon C. , Jodl Kathleen M. , Eccles Jaquelynne S.
Published: January 2004

The quality of father‐adolescent relationships, especially for nontraditional fathers, has been neglected in investigating adolescents’ beliefs. Closeness of father‐adolescent relationships was examined as a predictor of adolescents’ attitudes toward divorce. A sample of European and African American adolescents (N = 300) reported on the quality of father‐adolescent relationships in 11th grade and their attitudes toward divorce at age 19. Boys who felt close to their biological custodial fathers, biological noncustodial fathers, and stepfathers felt less likely to divorce in the future than boys who did not feel close to their fathers. The same was not true for girls. Feeling close to a father—regardless of father type—is associated with boys’ confidence in the stability of their future marriages.
Dollars, Dependency, and Divorce: Four Perspectives on the Role of Wives’ Income
Authors: Rogers Stacy J.
Published: January 2004

This article delineates and assesses the evidence for four perspectives that have guided previous research on the relationship between wives’ economic resources and marital stability. Hypotheses from these perspectives were tested using event history methods and 1980–1997 panel data for 1,704 individuals from the Marital Instability Over the Life Course study. Both wives’ dollar income and wives’ percentage of total family income served as measures of wives’ economic resources. The association between wives’ percentage of income and divorce formed an inverted U‐shaped curve, with the odds of divorce being highest when wives contributed between approximately 40% and 50% of the total family income. When wives’ resources were measured in dollars, wives’ income showed a positive, linear association with the odds of divorce. These findings provide the strongest support for an equal dependence perspective, which argues that economic dependence and obligation influence marital stability.
Interactions Between Cultural and Economic Determinants of Divorce in The Netherlands
Authors: Kalmijn Matthijs , De Graaf Paul M. , Poortman Anne‐Rigt
Published: January 2004

This study examines the relationship between gendered family roles and divorce in The Netherlands. Cultural and economic aspects of this relationship are distinguished. Economic hypotheses argue that the likelihood of divorce is increased if women work for pay and have attractive labor market resources. Cultural hypotheses argue that divorce chances are increased if women adhere to emancipatory norms, independent of their labor market positions. An event‐history analysis of a life‐history survey among 1,289 Dutch women reveals evidence for both hypotheses. Interaction effects are found as well: The protective effect of a traditional division of paid labor is only present among couples in which wives have traditional gender attitudes. Hence, the validity of economic explanations of divorce is conditional on cultural values.
Did Divorces Decline After the Oklahoma City Bombing?
Authors: Nakonezny Paul A. , Reddick Rebecca , Rodgers Joseph Lee
Published: January 2004

The Oklahoma City bombing in April 1995 was an act of terrorism that had many potential influences on the city and state, including influences on families. We analyzed divorce data from 1985 to 2000 for all 77 counties in Oklahoma to assess the divorce response to the Oklahoma City bombing. Our prediction was that divorce rates in Oklahoma would decrease in response to the bombing, a prediction derived from two different theoretical orientations, terror management theory and attachment theory. We test this prediction in the context of two relatively powerful quasi‐experimental designs, an interrupted time series design, and a nonequivalent control group design. We analyze the time series data with polynomial regression. Results suggested that there were lower observed divorce rates following the Oklahoma City bombing than the prevailing 10‐year cubic divorce trend would have predicted, with the effect dampening over time. We analyzed the nonequivalent control group design using an analysis of covariance (ANCOVA) linear model to compare counties in and around Oklahoma City to those farther away, and to compare metropolitan to nonmetropolitan counties in Oklahoma. In each case, divorces were lower in the predicted directions.
Fuel selection during intense shivering in humans: EMG pattern reflects carbohydrate oxidation
Authors: Haman François , Legault Stéphane R. , Weber Jean‐Michel
Published: January 2004

The thermogenic response of humans depends critically on the coordination of muscle fibre recruitment and oxidative fuel metabolism. The primary goal of this study was to determine whether the electromyographic (EMG) pattern of muscle recruitment could provide metabolic information on oxidative fuel selection during high‐intensity shivering. EMG activity (of 8 large muscles) and fuel metabolism were monitored simultaneously in non‐acclimatized adult men during high‐intensity shivering. Even though acute cold exposure elicited similar changes in metabolic rate among subjects, lipid and carbohydrate use was very different. Depending on the subject, the cold‐induced increase in carbohydrate (CHO) oxidation ranged between 2‐ and 8‐fold, with CHO accounting for 33–78% of total heat production , and lipids for 14–60%. This high variability in fuel selection was primarily explained by differences in ‘burst shivering’ rate, indicating that the recruitment of type II fibres plays a key role in orchestrating fuel selection. This study is the first to show that the pattern of muscle recruitment can provide quantitative information on energy metabolism. Future work should focus on the study of shivering bursts that may provide essential clues on what limits human survival in the cold.
Recruitment of Replication Protein A by the Papillomavirus E1 Protein and Modulation by Single-Stranded DNA
Authors: Loo Yueh-Ming , Melendy Thomas
Published: January 2004

With the exception of viral proteins E1 and E2, papillomaviruses depend heavily on host replication machinery for replication of their viral genome. E1 and E2 are known to recruit many of the necessary cellular replication factors to the viral origin of replication. Previously, we reported a physical interaction between E1 and the major human single-stranded DNA (ssDNA)-binding protein, replication protein A (RPA). E1 was determined to bind to the 70-kDa subunit of RPA, RPA70. In this study, using E1-affinity coprecipitation and enzyme-linked immunosorbent assay-based interaction assays, we show that E1 interacts with the major ssDNA-binding domain of RPA. Consistent with our previous report, no measurable interaction between E1 and the two smaller subunits of RPA was detected. The interaction of E1 with RPA was substantially inhibited by ssDNA. The extent of this inhibition was dependent on the length of the DNA. A 31-nucleotide (nt) oligonucleotide strongly inhibited the E1-RPA interaction, while a 16-nt oligonucleotide showed an intermediate level of inhibition. In contrast, a 10-nt oligonucleotide showed no observable effect on the E1-RPA interaction. This inhibition was not dependent on the sequence of the DNA. Furthermore, ssDNA also inhibited the interaction of RPA with papillomavirus E2, simian virus 40 T antigen, human polymerase alpha-primase, and p53. Taken together, our results suggest a potential role for ssDNA in modulating RPA-protein interactions, in particular, the RPA-E1 interactions during papillomavirus DNA replication. A model for recruitment of RPA by E1 during papillomavirus DNA replication is proposed.
Fusion to C3d Enhances the Immunogenicity of the E2 Glycoprotein of Type 2 Bovine Viral Diarrhea Virus
Authors: Wang Lingshu , Sunyer J. Oriol , Bello Leonard J.
Published: January 2004

The use of DNA and protein subunit vaccines in animals provides an opportunity to introduce vaccines that are arguably the safest that can be developed. For that reason, considerable effort is under way to devise methods of enhancing the immunogenicity of such vaccines. Seven years ago it was shown that fusing complement fragment C3d to hen egg lysozyme (HEL) enhanced the immunogenicity of HEL 10,000-fold. Based on this observation, we decided to evaluate the effect of C3d on the immunogenicity of the E2 protein of bovine viral diarrhea virus (BVDV). E2 is the major target of neutralizing antibody during BVDV infection. To test the effect of C3d on E2 immunogenicity, expression cassettes encoding a secreted form of E2 alone (E2s) or E2 fused to three copies of murine C3d (E2s-C3d) were constructed. The proteins were purified from the supernatants of transfected cells and used to immunize mice. The immune response was monitored by an enzyme-linked immunosorbent assay (ELISA) for E2s-specific antibody and by a virus neutralization test. The ELISA results indicated that the E2s-C3d protein is 10,000-fold more immunogenic than the E2s protein alone. The maximum primary immune response was elicited with <0.1 μg of E2s-C3d protein without an adjuvant. In addition, we have shown for the first time that high levels of anti-E2s and neutralizing antibodies can be elicited when this same low concentration of E2s-C3d is used to both prime and boost the immune response. We conclude that the E2s-C3d fusion protein has significant potential as a subunit vaccine against BVDV infection.
Innate Cellular Response to Virus Particle Entry Requires IRF3 but Not Virus Replication
Authors: Collins Susan E. , Noyce Ryan S. , Mossman Karen L.
Published: January 2004

Mammalian cells respond to virus infections by eliciting both innate and adaptive immune responses. One of the most effective innate antiviral responses is the production of alpha/beta interferon and the subsequent induction of interferon-stimulated genes (ISGs), whose products collectively limit virus replication and spread. Following viral infection, interferon is produced in a biphasic fashion that involves a number of transcription factors, including the interferon regulatory factors (IRFs) 1, 3, 7, and 9. In addition, virus infection has been shown to directly induce ISGs in the absence of prior interferon production through the activation of IRF3. This process is believed to require virus replication and results in IRF3 hyperphosphorylation, nuclear localization, and proteasome-mediated degradation. Previously, we and others demonstrated that herpes simplex virus type 1 (HSV-1) induces ISGs and an antiviral response in fibroblasts in the absence of both interferon production and virus replication. In this report, we show that the entry of enveloped virus particles from diverse virus families elicits a similar innate response. This process requires IRF3, but not IRF1, IRF7, or IRF9. Following virus replication, the large DNA viruses HSV-1 and vaccinia virus effectively inhibit ISG mRNA accumulation, whereas the small RNA viruses Newcastle disease virus, Sendai virus, and vesicular stomatitis virus do not. In addition, we found that IRF3 hyperphosphorylation and degradation do not correlate with ISG and antiviral state induction but instead serve as a hallmark of productive virus replication, particularly following a high-multiplicity infection. Collectively, these data suggest that virus entry triggers an innate antiviral response mediated by IRF3 and that subsequent virus replication results in posttranslational modification of IRF3, such as hyperphosphorylation, depending on the nature of the incoming virus.
CD46 Is a Cellular Receptor for Bovine Viral Diarrhea Virus
Authors: Maurer Karin , Krey Thomas , Moennig Volker , Thiel Heinz-Jürgen , Rümenapf Till
Published: January 2004

Various monoclonal antibodies (MAbs) that recognize cell surface proteins on bovine cells were previously shown to efficiently block infection with bovine viral diarrhea virus (BVDV) (C. Schelp, I. Greiser-Wilke, G. Wolf, M. Beer, V. Moennig, and B. Liess, Arch. Virol.140:1997-2009, 1995). With one of these MAbs, a 50- to 58-kDa protein was purified from calf thymus by immunoaffinity chromatography. Microchemical analysis of two internal peptides revealed significant sequence homology to porcine and human CD46. The cDNA of bovine CD46 (CD46bov) was cloned and further characterized. Heterologously expressed CD46bovwas detected by the MAb used for purification. A putative function of CD46bovas a BVDV receptor was studied with respect to virus binding and susceptibility of nonpermissive cells. While the expression of CD46bovcorrelated well with the binding of [3H]uridine-labeled BVDV, the susceptibility of cells nonpermissive for BVDV was not observed. However, the expression of CD46bovresulted in a significant increase in the susceptibility of porcine cells to BVDV. These results provide strong evidence that CD46bovserves as a cellular receptor for BVDV.
Small Internal Deletions in the Human Cytomegalovirus IE2 Gene Result in Nonviable Recombinant Viruses with Differential Defects in Viral Gene Expression
Authors: White Elizabeth A. , Clark Charles L. , Sanchez Veronica , Spector Deborah H.
Published: January 2004

The human cytomegalovirus (HCMV) IE2 86-kDa protein is a key viral transactivator and an important regulator of HCMV infections. We used the HCMV genome cloned as a bacterial artificial chromosome (BAC) to construct four HCMV mutants with disruptions in regions of IE2 86 that are predicted to be important for its transactivation and autoregulatory functions. Three of these mutants have mutations that remove amino acids 356 to 359, 427 to 435, and 505 to 511, which disrupts a region of IE2 86 implicated in the activation of HCMV early promoters, a predicted zinc finger domain, and a putative helix-loop-helix motif, respectively, while the fourth carries three arginine-to-alanine substitution mutations in the region of amino acids 356 to 359. The resulting recombinant viruses are not viable, and by using quantitative real-time reverse transcription-PCR and immunofluorescence we have determined the location of the block in their replicative cycles. The IE2 86Δ356-359 mutant is able to support early gene expression, as indicated by the presence of UL112-113 transcripts and UL112-113 and UL44 proteins in cells transfected with the mutant BAC. This mutant does not express late genes and behaves nearly indistinguishably from the IE2 86R356/7/9A substitution mutant. Both exhibit detectable upregulation of major immediate-early transcripts at early times. The IE2 86Δ427-435 and IE2 86Δ505-511 recombinant viruses do not activate the early genes examined and are defective in repression of the major immediate-early promoter. These two mutants also induce the expression of selected delayed early (UL89) and late genes at early times in the infection. We conclude that these three regions of IE2 86 are necessary for productive infections and for differential control of downstream viral gene expression.
Generation of High-Yielding Influenza A Viruses in African Green Monkey Kidney (Vero) Cells by Reverse Genetics
Authors: Ozaki Hiroichi , Govorkova Elena A. , Li Chenghong , Xiong Xiaoping , Webster Robert G. , Webby Richard J.
Published: January 2004

Influenza A viruses are the cause of annual epidemics of human disease with occasional outbreaks of pandemic proportions. The zoonotic nature of the disease and the vast viral reservoirs in the aquatic birds of the world mean that influenza will not easily be eradicated and that vaccines will continue to be needed. Recent technological advances in reverse genetics methods and limitations of the conventional production of vaccines by using eggs have led to a push to develop cell-based strategies to produce influenza vaccine. Although cell-based systems are being developed, barriers remain that need to be overcome if the potential of these systems is to be fully realized. These barriers include, but are not limited to, potentially poor reproducibility of viral rescue with reverse genetics systems and poor growth kinetics and yields. In this study we present a modified A/Puerto Rico/8/34 (PR8) influenza virus master strain that has improved viral rescue and growth properties in the African green monkey kidney cell line, Vero. The improved properties were mediated by the substitution of the PR8 NS gene for that of a Vero-adapted reassortant virus. The Vero growth kinetics of viruses with H1N1, H3N2, H6N1, and H9N2 hemagglutinin and neuraminidase combinations rescued on the new master strain were significantly enhanced in comparison to those of viruses with the same combinations rescued on the standard PR8 master strain. These improvements pave the way for the reproducible generation of high-yielding human and animal influenza vaccines by reverse genetics methods. Such a means of production has particular relevance to epidemic and pandemic use.
The Influenza B Virus Nonstructural NS1 Protein Is Essential for Efficient Viral Growth and Antagonizes Beta Interferon Induction
Authors: Dauber Bianca , Heins Gudrun , Wolff Thorsten
Published: January 2004

We analyzed the functions of the influenza B virus nonstructural NS1-B protein, both by utilizing a constructed mutant virus (ΔNS1-B) lacking the NS1 gene and by testing the activities of the protein when expressed in cells. The mutant virus replicated to intermediate levels in 6-day-old embryonated chicken eggs that contain an immature interferon (IFN) system, whereas older eggs did not support viral propagation to a significant extent. The ΔNS1-B virus was a substantially stronger inducer of beta IFN (IFN-β) transcripts in human lung epithelial cells than the wild type, and furthermore, transiently expressed NS1-B protein efficiently inhibited virus-dependent activation of the IFN-β promoter. Interestingly, replication of the ΔNS1-B knockout virus was attenuated by more than 4 orders of magnitude in tissue culture cells containing or lacking functional IFN-α/β genes. These findings show that the NS1-B protein functions as a viral IFN antagonist and indicate a further requirement of this protein for efficient viral replication that is unrelated to blocking IFN effects.
Large-Scale Identification of Disease Genes Involved in Acute Myeloid Leukemia
Authors: Erkeland Stefan J. , Valkhof Marijke , Heijmans-Antonissen Claudia , van Hoven-Beijen Antoinette , Delwel Ruud , Hermans Mirjam H. A. , Touw Ivo P.
Published: January 2004

Acute myeloid leukemia (AML) is a heterogeneous group of diseases in which chromosomal aberrations, small insertions or deletions, or point mutations in certain genes have profound consequences for prognosis. However, the majority of AML patients present without currently known genetic defects. Retroviral insertion mutagenesis in mice has become a powerful tool for identifying new disease genes involved in the pathogenesis of leukemia and lymphoma. Here we have used the Graffi-1.4 strain of murine leukemia virus, which causes predominantly AML, in a screen to identify novel genes involved in the pathogenesis of this disease. We report 79 candidate disease genes in common integration sites (CISs) and 15 genes whose family members previously were found to be affected in other studies. The majority of the identified sequences (60%) were not found in lymphomas and monocytic leukemias in previous screens, suggesting a specific involvement in AML. Although most of the virus integrations occurred in or near the 5′ or 3′ ends of the genes, suggesting deregulation of gene expression as a consequence of virus integration, 18 CISs were located exclusively within the genes, conceivably causing gene disruption.
Discrete Domains within the Rotavirus VP5
*
Direct Peripheral Membrane Association and Membrane Permeability
Authors: Golantsova Nina E. , Gorbunova Elena E. , Mackow Erich R.
Published: January 2004

Cleavage of the rotavirus spike protein, VP4, is required for rotavirus-induced membrane permeability and viral entry into cells. The VP5
*
cleavage product selectively permeabilizes membranes and liposomes and contains an internal hydrophobic domain that is required for membrane permeability. Here we investigate VP5
*
domains (residues 248 to 474) that direct membrane binding. We determined that expressed VP5 fragments containing residues 248 to 474 or 265 to 474, including the internal hydrophobic domain, bind to cellular membranes but are not present in Triton X-100-resistant membrane rafts. Expressed VP5 partitions into aqueous but not detergent phases of Triton X-114, suggesting that VP5 is not integrally inserted into membranes. Since high-salt or alkaline conditions eluted VP5 from membranes, our findings demonstrate that VP5 is peripherally associated with membranes. Interestingly, mutagenesis of residue 394 (W→R) within the VP5 hydrophobic domain, which abolishes VP5-directed permeability, had no effect on VP5's peripheral membrane association. In contrast, deletion of N-terminal VP5 residues (residues 265 to 279) abolished VP5 binding to membranes. Alanine mutagenesis of two positively charged residues within this domain (residues 274R and 276K) dramatically reduced (>95%) binding of VP5 to membranes and suggested their potential interaction with polar head groups of the lipid bilayer. Mutations in either the VP5 hydrophobic or basic domain blocked VP5-directed permeability of cells. These findings indicate that there are at least two discrete domains within VP5
*
required for pore formation: an N-terminal basic domain that permits VP5
*
to peripherally associate with membranes and an internal hydrophobic domain that is essential for altering membrane permeability. These results provide a fundamental understanding of interactions between VP5
*
and the membrane, which are required for rotavirus entry.
Complete Genome Sequence of Fer-de-Lance Virus Reveals a Novel Gene in Reptilian Paramyxoviruses
Authors: Kurath Gael , Batts William N. , Ahne Winfried , Winton James R.
Published: January 2004

The complete RNA genome sequence of the archetype reptilian paramyxovirus, Fer-de-Lance virus (FDLV), has been determined. The genome is 15,378 nucleotides in length and consists of seven nonoverlapping genes in the order 3′ N-U-P-M-F-HN-L 5′, coding for the nucleocapsid, unknown, phospho-, matrix, fusion, hemagglutinin-neuraminidase, and large polymerase proteins, respectively. The gene junctions contain highly conserved transcription start and stop signal sequences and tri-nucleotide intergenic regions similar to those of other
Paramyxoviridae
. The FDLV P gene expression strategy is like that of rubulaviruses, which express the accessory V protein from the primary transcript and edit a portion of the mRNA to encode P and I proteins. There is also an overlapping open reading frame potentially encoding a small basic protein in the P gene. The gene designated U (unknown), encodes a deduced protein of 19.4 kDa that has no counterpart in other paramyxoviruses and has no similarity with sequences in the National Center for Biotechnology Information database. Active transcription of the U gene in infected cells was demonstrated by Northern blot analysis, and bicistronic N-U mRNA was also evident. The genomes of two other snake paramyxovirus genotypes were also found to have U genes, with 11 to 16% nucleotide divergence from the FDLV U gene. Pairwise comparisons of amino acid identities and phylogenetic analyses of all deduced FDLV protein sequences with homologous sequences from other
Paramyxoviridae
indicate that FDLV represents a new genus within the subfamily
Paramyxovirinae
. We suggest the name Ferlavirus for the new genus, with FDLV as the type species.
Cytotoxic T Lymphocytes Derived from Patients with Chronic Hepatitis C Virus Infection Kill Bystander Cells via Fas-FasL Interaction
Authors: Gremion Christel , Grabscheid Benno , Wölk Benno , Moradpour Darius , Reichen Jürg , Pichler Werner , Cerny Andreas
Published: January 2004

The role of Fas-mediated lysis of hepatocytes in hepatitis C virus (HCV)-induced injury is frequently discussed. We therefore analyzed the effect of the number of HCV antigen-expressing cells, the mode of antigen presentation, and the number of cytotoxic T lymphocytes in a coculture system mimicking cellular components of the liver. Here, we show that endogenously processed HCV proteins are capable of inducing bystander killing. We further demonstrate that 0.8 to 1.5% of cells presenting HCV antigens suffice to induce lysis of 10 to 29% of bystander cells, suggesting that the mechanism may be operative at low fractions of infected versus uninfected hepatocytes in vivo. Our data underscore the role of the Fas pathway in HCV-related liver injury and support the exploration of Fas-based treatment strategies for patients with chronic hepatitis C virus infection.
Inactivation of Transmissible Spongiform Encephalopathy (Prion) Agents by Environ LpH
Authors: Race Richard E. , Raymond Gregory J.
Published: January 2004

Agents causing transmissible spongiform encephalopathy (TSE) diseases are resistant to inactivation by several conventional decontamination methods. Using an animal bioassay, we compared the TSE agent disinfectant efficacy of a commercially available product referred to alternatively as LpH-SE, LpH-AG, or LpH-st to that of a similarly named but differently formulated product, Environ LpH, which was found to be an effective TSE agent disinfectant in a previous study. Here, we found LpH-SE to be at least 10
4
-fold to 10
5
-fold less effective than Environ LpH.
Intermolecular Interactions in the (CO2)2, N2-CO2 and CO-CO2 Complexes
Authors: Fišer Jiří , Boublík Tomáš , Polák Rudolf
Published: January 2004

Interaction energies of the most stable structures of the title complexes are calculated using the supermolecule CCSD(T) and MP4 methods and aug-cc-pVXZ (X = D, T, Q) basis sets extended by a set of midbond functions centered in the middle of the intermolecular bond. Geometrical parameters for these structures are in very good agreement with experimental data.
Additivity of Absolute Entropies
Authors: Černušák Ivan , Gregurick Susan K. , Roswell Marjorie , Deakyne Carol A. , Jenkins H. Donald Brooke , Liebman Joel F.
Published: January 2004

Symmetry number-corrected entropies for a series of inorganic diatomic molecules were analyzed. We have considered molecules XY (X, Y = H, F, Cl, Br, I, Li, Na, K, Rb, Cs, B and Al). Based on the assumption of elemental additivity, the prediction of known entropy data is quite satisfactory (except for H2) and provides fairly reasonable estimates for data currently unavailable.
Quantum-Chemical Insight Into Mechanism of Combined Intra-Intermolecular Cycloaddition
Authors: Kulhánek Petr , Potáček Milan , Koča Jaroslav
Published: January 2004

Two different reaction mechanisms of a mixed criss-cross cycloaddition with opposite sequence of reaction steps, intra-intermolecular and inter-intramolecular, were explored by quantum-chemical calculations at the MP2 and B3LYP levels of theory and with cc-pVDZ basis set. It was found that the rate-determining step in the both mechanisms is the first step regardless of the mechanism. The Gibbs activation barrier of the intramolecular step of the intra-intermolecular sequence is by 10.4 kcal mol-1 lower than that of the intermolecular step of the inter-intramolecular sequence at the MP2 level of theory and almost the same at the B3LYP level of theory. This together with known experimental data confirms that the intra-intermolecular sequence is the most probable mechanism.
Non-neoplastic granulosa cells within ovarian vascular channels: a rare potential diagnostic pitfall
Authors: McCluggage W G , Young R H
Published: January 2004

Aims:To describe six cases seen in consultation in which artefactual vascular involvement within the ovary by benign granulosa cells caused diagnostic confusion.

Methods/Results:In five cases, the initial favoured diagnoses of the submitting pathologists were metastatic carcinoma (three cases) and immature neural elements within a teratoma (two cases). In two cases, the ovary contained a benign cystic teratoma (one with struma ovarii), in two cases endometriosis, in one case follicular cysts, and in the other no pathological lesion was present. In all cases, several small ovarian vascular channels contained cohesive groups of cells with mildly atypical nuclei and cytoplasm, which varied from scant to abundant and eosinophilic. In four cases, mitotic figures were identified. The cells were morphologically consistent with benign granulosa cells and were associated in four cases with a nearby follicle lined by similar cells. There was no evidence of a mass lesion, grossly or histologically, to suggest a granulosa cell tumour. The nature of the cells was confirmed using immunohistochemistry for α inhibin and calretinin in one case.

Conclusions:This phenomenon is probably an artefact secondary to surgical trauma or sectioning within the laboratory; alternatively, it could be related to ovulation. It is important that this benign process is not misinterpreted as cancer, either primary or metastatic, which may prompt inappropriate treatment or investigations that are not needed.
A follow up model for patients with atrophic chronic gastritis and intestinal metaplasia
Authors: Dinis-Ribeiro M , Lopes C , da Costa-Pereira A , Guilherme M , Barbosa J , Lomba-Viana H , Silva R , Moreira-Dias L
Published: January 2004

Aim:To devise a follow up model for patients with gastric cancer associated lesions, such as atrophic chronic gastritis (ACG) and intestinal metaplasia (IM).

Methods:Cohort study of 144 patients, followed for a minimum of one year, in whom at least two upper gastrointestinal endoscopic biopsies in flat gastric mucosa provided a diagnosis of ACG, IM, or low grade dysplasia (LGD).

Results:Of those diagnosed with ACG at first endoscopic biopsy (entry biopsy), 12% progressed to LGD in outcome biopsy, as did 8% of those with type I IM, 38% with type II or III IM, and 32% with LGD. Type of IM at entry independently predicted progression to LGD and cancer. Type II and III IM had a higher rate of progression to LGD than type I IM, which showed an indolent behaviour similar to ACG. Patients with type II or III IM were at higher risk for development of dysplasia, and 7% of patients with type III IM at first biopsy progressed to high grade dysplasia (HGD), whereas no cases of ACG or type I/II IM progressed to HGD during the first three years.

Conclusion:Patients with ACG or IM could possibly be allocated to different management schedules, based on differences in rate and proportion of progression to LGD or HGD. Less intensive follow up (two/three yearly with “serological evaluation” (pepsinogen)) may suit those with ACG or type I IM. Patients with type III IM may benefit from six to 12 monthly improved endoscopic examination (magnification chromoendoscopy).
The immunohistochemical expression of calcitonin receptor-like receptor (CRLR) in human gliomas
Authors: Benes L , Kappus C , McGregor G P , Bertalanffy H , Mennel H D , Hagner S
Published: January 2004

Background:Gliomas are the most common primary tumours of the central nervous system and exhibit rapid growth that is associated with neovascularisation. Adrenomedullin is an important tumour survival factor in human carcinogenesis. It has growth promoting effects on gliomas, and blockade of its actions has been experimentally shown to reduce the growth of glioma tissues and cell lines. There is some evidence that the calcitonin receptor-like receptor (CRLR) mediates the tumorigenic actions of adrenomedullin.

Aim:To determine whether CRLR is expressed in human gliomas and the probable cellular targets of adrenomedullin.

Methods:Biopsies from 95 human gliomas of varying grade were processed for immunohistochemical analysis using a previously developed and characterised antibody to CRLR.

Results:All tumour specimens were positive for CRLR. As previously found in normal peripheral tissues, CRLR immunostaining was particularly intense in the endothelial cells. This was evident in all the various vascular conformations that were observed, and which are typical of gliomas. In addition, clear immunostaining of tumour cells with astrocyte morphology was observed. These were preferentially localised around vessels.

Conclusions:This study has shown for the first time that the CRLR protein is present in human glioma tissue. The expression of the receptor in endothelial cells and in astrocytic tumour cells is consistent with the evidence that its endogenous ligand, adrenomedullin, may influence glioma growth by means of both direct mitogenic and indirect angiogenic effects. CRLR may be a valuable target for effective therapeutic intervention in these malignant tumours.
The effect of a symptom related “gating policy” on ANCA requests in routine clinical practice
Authors: Sinclair D , Saas M , Stevens J M
Published: January 2004

Background:Most positive antineutrophil cytoplasmic antibody (ANCA) results are associated with non-vasculitic conditions, and guidelines have been proposed for the judicious use of this test. The outcome of applying similar guidelines in a routine laboratory is reported.

Methods:All immunology requests (6500) over six months were selected, and those requesting ANCA were studied for the appropriateness of the clinical data supporting the request, the presence of ANCA in those samples tested, and the final diagnosis. Antibodies were detected by indirect immunofluorescence.

Results:ANCA testing was requested in 287 samples. Application of a “gating policy”, which refuses analysis on requests that are not supported by clinical data suggestive of systemic vasculitis, made clinicians more selective about the patients for whom they requested ANCA testing. The percentage of “appropriate” screens for systemic vasculitis was relatively high (212 of 287 requests: 72.5%). Only one of the remainder, for whom ANCA testing was initially refused, developed an ANCA related systemic vasculitis in the two years after the study, but the delay in reporting her positive ANCA was only two days. Most of the samples tested were negative (155 of 212), but most (42 of 57) of the patients with positive ANCA results were found to have a systemic vasculitis.

Conclusions:A gating policy to select requests supported by clinical data suggestive of systemic vasculitis makes ANCA testing more clinically relevant and cost effective. Studies where guidelines can be proposed and their effects measured are important in the light of clinical governance and evidence based medicine.
Infection of the CNS byScedosporium apiospermumafter near drowning. Report of a fatal case and analysis of its confounding factors
Authors: Kowacs P A , Soares Silvado C E , Monteiro de Almeida S , Ramos M , Abrão K , Madaloso L E , Pinheiro R L , Werneck L C
Published: January 2004

This report describes a fatal case of central nervous system pseudallescheriasis. A 32 year old white man presented with headache and meningismus 15 days after nearly drowning in a swine sewage reservoir. Computerised tomography and magnetic resonance imaging of the head revealed multiple brain granulomata, which vanished when steroid and broad spectrum antimicrobial and antifungal agents, in addition to dexamethasone, were started. Cerebrospinal fluid analysis disclosed a neutrophilic meningitis. Treatment with antibiotics and amphotericin B, together with fluconazole and later itraconazole, was ineffective. Miconazole was added through an Ommaya reservoir, but was insufficient to halt the infection.Pseudallescheria boydiiwas finally isolated and identified in cerebrospinal fluid cultures, a few days before death, three and a half months after the symptoms began. Diagnosis was delayed because of a reduction in the lesions after partial treatment, which prevented a stereotactic biopsy. Physicians should be aware of this condition, and provide prompt stereotactic biopsy. Confirmed cases should perhaps be treated with voriconazole, probably the most effective, currently available treatment for this agent.