bluuebunny/crossref_metadata_2025 huggingface dataset:
Results of a hepatitis C general transfusion lookback program for patients who received blood products before July 1992
Authors: Williams James L. , Cagle Henry H , Christensen Carol J. , Fox‐Leyva Leslie K. , McMahon Brian J.
Published: June 2005

BACKGROUND: The Centers for Disease Control and Prevention recommend hepatitis C virus (HCV) antibody (anti‐HCV) screening for persons who received blood products before July 1992. A general transfusion lookback program was implemented to identify, counsel, and screen persons who received transfusions at the Alaska Native Medical Center between January 1980 and July 1992.

STUDY DESIGN AND METHODS: Hard‐copy transfusion records data were entered, and available databases were queried to identify deceased patients and the mailing address of those living. Patients were notified by letter of their HCV risk and encouraged to seek counseling and testing. Serum samples were screened for anti‐HCV and HCV RNA, and program costs were estimated.

RESULTS: Overall, 3169 transfusion recipients were identified, with 1356 (43%) living and targeted for notification. Of 764 patients notified and screened by this program, 41 (5%) were anti‐HCV–positive and 19 (2%) were HCV RNA–positive. There was a higher probability of detecting anti‐HCV with each subsequent increase of a transfusion event. Among 298 lookback patients, 33 percent were unaware of having received a blood transfusion. The estimated cost per person sent notification was US$57 and to detect an anti‐HCV–positive case it was US$3146.

CONCLUSION: This general transfusion lookback program successfully notified and screened patients at a reasonable cost. Further investigation would be helpful in determining the role these programs or other measures could play in promoting HCV screening in persons receiving transfusions before July 1992, especially among those who are unaware of their transfusion history.
Onset of expression of the components of the Kell blood group complex
Authors: Pu Jeffrey J. , Redman Colvin M. , Visser Jan W.M. , Lee Soohee
Published: June 2005

BACKGROUND:Kell and XK, two distinct red blood cell membrane proteins, are linked by a disulfide bond and form the Kell blood group complex. Kell surface antigens are expressed early during erythropoiesis but the onset of expression of XK which carries the Kx antigen is unknown.

STUDY DESIGN AND METHODS:To determine whether Kell and XK are synchronously expressed, sorted human hematopoietic progenitor cells and mouse progenitor cells of defined lineage were studied. To determine the onset of expression, human marrow and cord blood cells were sorted into three subpopulations, representing stem, multipotent, and erythroid progenitor cells, and the expression of Kell and XK was determined by reverse transcription–polymerase chain reaction (RT‐PCR) and fluorescence‐activated cell sorting (FACS) analysis. Mouse Kell and XK transcripts were determined by cDNA blotting of progenitor cells of defined lineage.

RESULTS:By RT‐PCR, human peripheral blood progenitor cells had weak expression of Kell and XK transcripts but FACS analysis did not detect surface antigens. Kell and XK transcripts are expressed in multipotent progenitor cells and these cells express Kell surface antigens. The expression of Kx antigen in progenitor cells was not determined owing to nonspecific reactions with the antibody. By cDNA blotting, mouse Kell expression was detected in bipotential megakaryocytes‐erythroid cells and in colony‐forming units–erythroid (CFU–E) and burst‐forming units–erythroid (BFU–E), whereas XK was only detected in CFU‐E and BFU‐E.

CONCLUSION:Both Kell and XK transcripts occur early during erythropoiesis; however, expression may not be coincident because, in mice, Kell transcripts, but not XK, occur in bipotential megakaryocytes‐erythroid progenitor cells.
Placebo‐controlled study of intravenous magnesium supplementation during large‐volume leukapheresis in healthy allogeneic donors
Authors: Haddad Salim , Leitman Susan F. , Wesley Robert A. , Cecco Stacey , Yau Yu Ying , Starling Judith , Rehak Nadja N. , Bolan Charles D.
Published: June 2005

BACKGROUND: Marked decreases in ionized magnesium (iMg) levels occur during large‐volume leukapheresis (LVL); however, the effect of intravenous (IV) magnesium supplementation in this setting has not been carefully studied.

STUDY DESIGN AND METHODS: Thirty healthy allogeneic peripheral blood progenitor cell donors receiving citrate anticoagulant with IV calcium prophylaxis were randomized to receive either IV magnesium (0.2 mg Mg per mL acid citrate dextrose‐A) or placebo during LVL, with a double‐blind design.

RESULTS: Thirty subjects underwent 75 LVL pro‐ cedures, 37 with magnesium and 38 with placebo. Group characteristics were similar for sex, weight, citrate infusion rate (1.36 mg/kg/min vs. 1.37 mg/kg/min), and volume processed (16 L vs. 17 L). Serum iMg levels remained within the reference range with magnesium supplementation, but decreased 39 ± 11 percent below baseline (p < 10−10) after placebo, with greater decreases after consecutive procedures. Subjects receiving magnesium had more vigorous parathyroid hormone responses and higher glucose levels and also tended to have higher serum potassium and ionized calcium levels. Mild paresthesias, coldness, and nausea occurred in 28, 20, and 7 percent of donors, respectively, with no significant differences between groups. Severe symptoms (chest tightness) occurred in only one subject receiving placebo.

CONCLUSION: IV magnesium supplementation exerts a significant impact on serum magnesium levels, but does not reduce the frequency or severity of the relatively mild citrate‐related effects observed in LVL performed with continuous IV calcium prophylaxis.
Effects of a new pathogen‐reduction technology (Mirasol PRT) on functional aspects of platelet concentrates
Authors: Perez‐Pujol S. , Tonda R. , Lozano M. , Fuste B. , Lopez‐Vilchez I. , Galan A.M. , Li J. , Goodrich R. , Escolar G.
Published: June 2005

BACKGROUND: Several strategies are being developed to reduce the risk of pathogen transmission associated with platelet (PLT) transfusion.

STUDY DESIGN AND METHODS: The impact of a new technology for pathogen reduction based on riboflavin plus illumination (Mirasol PRT, Navigant Biotechnologies, Inc.) at 6.2 and 12.3 J per mL on functional and biochemical characteristics of PLTs was evaluated. PLT concentrates (PCs) obtained by apheresis were treated with Mirasol PRT and stored at 22°C. Modifications in major PLT glycoproteins (GPIbα, GPIV, and GPIIb‐IIIa), adhesive ligands (von Willebrand factor [VWF], fibrinogen [Fg], and fibronectin), activation antigens (P‐selectin and LIMP), and apoptotic markers (annexin V binding and factor [F]Va) were analyzed by flow cytometry. Adhesive and cohesive PLT functions were evaluated with well‐established perfusion models. Studies were performed on the preparation day (Day 0) and during PCs storage (Days 3 and 5).

RESULTS: Levels of glycoproteins remained stable during storage in PCs treated with 6.2 J per mL pathogen reduction technology (PRT) and similar to those observed in nontreated PCs. When 12.3 J per mL PRT was applied, however, levels of GPIbα moderately decreased on Days 3 and 5. VWF, Fg, and FVa were not modified in their expression levels, either by treatment or by storage period. Fibronectin appeared more elevated in all PRT samples. A progressive increase in P‐selectin and LIMP expression and in annexin V binding was observed during storage of PRT‐treated PCs. Functional studies indicated that 6.2 J per mL Mirasol PRT–treated PLTs preserved adhesive and cohesive functions to levels compatible with those observed in the respective control PCs.

CONCLUSION: PLT function was well preserved in PCs treated with 6.2 J per mL Mirasol PRT and stored for 5 days.
Enhancement of a culture‐based bacterial detection system (eBDS) for platelet products based on measurement of oxygen consumption
Authors: Holme Stein , McAlister Morven B. , Ortolano Girolamo A. , Chong Chiyong , Cortus Mary Anne , Jacobs Michael R. , Yomtovian Roslyn , Freundlich Lawrence F. , Wenz Barry
Published: June 2005

BACKGROUND: An enhanced bacterial detection system (Pall eBDS) was developed that distinguishes itself from its predecessor (Pall BDS) by removal of the platelet (PLT)‐retaining filter allowing for optimal bacterial transfer, modification of the culture tablet to reduce the confounding effects of respiring PLTs while enhancing bacterial growth, and facilitation of nutrients and gas exchange by agitating the sample pouch during incubation at 35°C. The objective was to evaluate the performance of the new eBDS.

STUDY DESIGN AND METHODS: Leukoreduced whole blood–derived PLT concentrates (LR‐PCs) and LR single‐donor PLTs (LR‐SDPs) were inoculated with 1 to 15 colony‐forming units (CFUs) of bacteria per mL in studies of each of 10 bacterial species associated with fatal transfusion‐transmitted bacterial infection. Immediately after inoculation and after 24 hours of storage at 22°C, samples of inoculated LR‐PCs were aseptically transferred into the eBDS pouches. Pouches were then incubated for 24 hours at 35°C with agitation and oxygen concentration was then measured.

RESULTS: Median inoculation levels ranged from 5 to 13 CFUs per mL for each species studied. No significant differences in oxygen concentration were found when comparing LR‐PCs with LR‐SDPs. When sampling occurred from the PLTs 24 hours after inoculation, all 280 cases (24‐33 replicates of each species) were detected as contaminated by the device (100% sensitivity). No false‐positives were obtained with 713 uninoculated PLT units.

CONCLUSIONS: The eBDS demonstrated improved detection sensitivity in the range of 1 to 15 CFUs per mL with no observed false‐positives compared to the original BDS (detection range 100 to 500 CFUs/mL) with no false‐positives.
Serum N‐Terminal pro‐BNP Levels Correlate with Symptoms and Echocardiographic Findings in Patients with Mitral Stenosis
Authors: Arat‐Özkan Alev , Kaya Ayşem , Yiǧit Zerrin , Balcı Huriye , Ökçün Bariş , Yazıcıoǧlu Nuran , Küçükoǧlu Serdar
Published: June 2005

This study is designed to evaluate the N‐terminal pro‐BNP (NTproBNP) levels in patients with mitral stenosis (MS) and its possible correlation with clinical and echocardiographic parameters of the disease. The study group consisted of 29 patients with isolated MS (patients with greater mild regurgitation were excluded) and 20 normal control subjects of similar age and gender distribution. Blood samples for NTproBNP were collected at the time of clinical and echocardiographic examination. NTproBNP levels were elevated in patients with MS compared to controls (325 ± 249 pg/dL [19.9–890] versus 43 ± 36 pg/dL [5.76–193.3], P < 0.001). Patients with atrial fibrillation had significantly higher NTproBNP levels compared to those with sinus rhythm (561 ± 281 pg/dL versus 254 ± 194 pg/dL, P = 0.044). MS patients with sinus rhythm also had higher NTproBNP levels compared to controls (254 ± 194 pg/dL versus 43 ± 36 pg/dL, P = 0.00011). NT pro BNP levels correlated to the LA (R = 0.73, P < 0.0001) and RV (R = 0.41, P = 0.042) diameters, mitral valve area (R =−0.45, P = 0.025), mean mitral gradient (R = 0.57, P = 0.003), peak PAP (R = 0.7, P = 0.03), and NYHA functional class (R = 0.61, P = 0.007). In conclusion, serum NTproBNP levels correlate well with echocardiographic findings and functional class in patients with MS and can be used as a marker of disease severity. Additionally, it may have a potential use as an additional noninvasive and relatively cheap method in monitoring disease progression especially in patients with poor echocardiographic windows.
Ontogeny of hematological cell and biochemical profiles in maternal and fetal baboons (Papio species)
Authors: Schlabritz‐Loutsevitch Natalia E. , Hubbard Gene B. , Jenkins Susan L. , Martin Helen C. , Snider Cathy S. , Frost Patrice A. , Michelle Leland M. , Havill Lorena M. , McDonald Thomas J. , Nathanielsz Peter W.
Published: June 2005

Abstract:  The normal ranges of hematological cell profiles and biochemistry are documented in adult non‐pregnant, pregnant, juvenile, and neonatal baboons. Despite the extensive use of the baboon as a model for the study of various aspects of pregnancy, there is no data from paired mothers and their fetuses at different stages of gestation. Hematologic and biochemical profile data were obtained from eight non‐pregnant female baboons, 37 mothers and 38 fetal baboons at 30 ± 2, 90 ± 2, 125 ± 2, and 175 ± 2 days of gestation (mean ± range; dGA; term, 180 dGA). Changes observed in fetal and maternal blood during normal baboon pregnancy were similar to those reported in human pregnancy. The level of alkaline phosphatase was two times higher in fetal blood circulation than that reported in human pregnancy.
Evaluation of a timed and repeated perianal tape test for the detection of pinworms (Trypanoxyuris microon) in owl monkeys (Aotus nancymae)
Authors: Felt S.A. , White C.E.
Published: June 2005

Abstract:  The purpose of this project was to determine if the detection of pinworm infections in owl monkeys (Aotus nancymae) could be improved by performing perianal tape testing at specific times of the day and/or by performing repeated sampling. Eight Aotus known to be infected with pinworms were sampled at four selected time points (06:00, 12:00, 18:00 and 24:00 hours) over the course of a 3‐week period. Samples were examined microscopically and oxyurid eggs were quantified. Results revealed no significant differences in time points, but did indicate that repeated sampling significantly improved pinworm egg detection. Results also determined that Aotus housed with an infected cagemate are at an approximately 14‐times greater risk of being infected than animals housed without an infected cagemate. Lastly, results indicated no significant difference between peripheral eosinophil and basophil numbers from infected and clean animals.
Nephroblastomatosis and nephroblastoma in nonhuman primates
Authors: Denise Goens S. , Moore Charleen M. , Brasky Kathleen M. , Frost Patrice A. , Michelle Leland M. , Hubbard Gene B.
Published: June 2005

Abstract:  Wilms’ tumors, or nephroblastomas, are renal embryonal malignancies with a high incidence in humans. Nephroblastomas are uncommon in nonhuman primates. This report describes three cases of spontaneous proliferative renal tumors in young monkeys: two cases of unilateral kidney nephroblastomas in baboons and a nephroblastomatosis in a cynomolgus macaque. Histologically, both baboon tumors were typical of Wilms’ tumors found in humans, with proliferative epithelial cells forming tubules and aborted glomeruli, nephrogenic rests and proliferative fibrovascular tissue. The left kidney of the macaque was markedly enlarged and histologically similar to the baboon tumors, although normal kidney architecture was completely effaced by primitive tubules and occasional glomeruli surrounded by edematous stromal tissue. Cytogenetic analysis did not detect any macaque or baboon equivalents to human Wilms’ tumor chromosomal abnormalities. By human pathology classification, the diffuse nature of the macaque tumor is more consistent with nephroblastomatosis than nephroblastoma. This differentiation is the first to be reported in a species other than human. The nephroblastomas described here are the first nephroblastomas to be reported in baboons. Our observations indicate that nonhuman primate nephroblastomatosis and nephroblastomas develop in a similar way to Wilms’ tumors in humans, although no genetic marker has been associated with nephroblastomas of nonhuman primates thus far.
Naturally occurring menopause in cynomolgus monkeys: changes in hormone, lipid, and carbohydrate measures with hormonal status
Authors: Kavanagh Kylie , Koudy Williams J. , Wagner Janice D.
Published: June 2005

Abstract:  Naturally occurring post‐menopausal (PM) female cynomolgus monkeys (Macaca fascicularis) were identified. Their sex hormone profile was characterized and compared with younger pre‐menopausal females before and after ovariectomy (OVX). PM females had lower estrogens and increased follicle‐stimulating hormone (FSH) concentrations. Two PM females had diabetes mellitus and elevated androgens (androstenodione and dihydroepiandrosterone sulfate). Non‐diabetic PM females were given parenteral E2 which normalized FSH, and caused improvements in body weight, plasma lipids and lipoprotein cholesterol. Androgens remained lower with E2 treatment. OVX induced comparable increases in FSH seen with the PM monkeys, however they had lower bodyweights, and had higher estrone and androstenodione concentrations. Natural menopause occurs in cynomolgus monkeys and hormone changes with OVX are similar however, differences in sex hormones that can relate to body mass and age may be important. E2 treatment restored estrogen levels and induced improvements in the lipid profile of PM females.
Pharmacokinetics and pharmacodynamics of injectable testosterone undecanoate in castrated cynomolgus monkeys (Macaca fascicularis) are independent of different oil vehicles
Authors: Wistuba Joachim , Marc Luetjens C. , Kamischke Axel , Gu Yi‐Qun , Schlatt Stefan , Simoni Manuela , Nieschlag Eberhard
Published: June 2005

Abstract:  Testosterone undecanoate (TU) dissolved in soybean oil was developed in China to improve the pharmacokinetics of this testosterone ester in comparison with TU in castor or tea seed oil. As a pre‐clinical primate model, three groups of five castrated cynomolgus macaques received either a single intramuscular injection of 10 mg/kg bodyweight TU in soybean oil, in tea seed oil, or in castor oil (equals 6.3 mg pure T/kg bodyweight for all preparations). Testosterone, estradiol, luteinizing hormone, and follicle‐stimulating hormone as well as prostate volume, body weight and ejaculate weight were evaluated. After injection supraphysiological testosterone levels were induced. There were no significant differences in the pharmacokinetics of the three TU preparations for testosterone and estradiol. The gonadotropin levels showed a high individual variation. Prostate volumes increased equally in all groups after administration and declined to castrate level afterwards. The results suggest that TU in soybean oil produces similar effects as TU in the other vehicles. This study in non‐human primates provides no objection to testing of this new preparation in humans.
Real‐time polymerase chain reaction quantification of Epstein–Barr virus in chronic periodontitis patients
Authors: Konstantinidis Antonis , Sakellari Dimitra , Papa Anna , Antoniadis Antonis
Published: June 2005

Background:  Although herpes viruses have been implicated in the pathogenesis of chronic and aggressive periodontitis, few data in the literature refer to quantification of these viruses in periodontal sites, especially in relation to serological findings.

Objective:  The aim of the present study was to compare Epstein–Barr virus (EBV) DNA load in subgingival specimens from chronic periodontitis patients and in periodontally healthy subjects, in relation to serologic testing of IgM and IgG antibodies to EBV.

Methods:  A total of 22 chronic periodontitis patients and 13 controls participated in the present study. Seventy‐nine subgingival specimens (one pooled, one from a deep and one from a shallow site), sampled with paper points, were analysed with real‐time polymerase chain reaction for EBV. Subjects were also examined for anti‐EBV IgG and IgM levels in serum, using an enzyme‐linked immunosorbent assay.

Results:  One subject was seronegative for EBV. Three subjects (one patient and two controls) displayed anti‐EBV IgM. Their data were excluded from further analysis. All three displayed EBV in their subgingival samples. Nine out of the remaining 20 chronic periodontitis patients and 10 out of 11 controls did not display EBV subgingivally. A statistically significant difference in viral load was observed between pooled and shallow‐pocket samples from periodontitis patients but not between samples from deep and shallow pockets (Kruskall–Wallis anova, Dunn's multiple comparisons test).

Conclusions:  Data from the present study do not strongly support the pathogenetic significance of EBV in chronic periodontitis lesions. The data do, however, suggest that parallel serological assessments provide a useful insight into the association of viruses with periodontal disease.
A functional interleukin‐1β gene polymorphism is associated with chronic periodontitis in a sample of Brazilian individuals
Authors: Moreira Paula R. , De Sá Alessandra R. , Xavier Guilherme M. , Costa José E. , Gomez Ricardo S. , Gollob Kenneth J. , Dutra Walderez O.
Published: June 2005

Background:  Interleukin‐1 beta (IL‐1β) is a potent inflammatory mediator and an important polymorphism in the locus +3954 (C/T) of the human IL1B gene has been shown to affect the levels of this cytokine. This functional polymorphism has been associated with the establishment of inflammatory diseases, including periodontal disease, in European, Asian and North American populations.

Objective:  The aim of this study was to investigate the association between the IL1B (+3954) gene polymorphism and the occurrence of different clinical forms of periodontitis in a sample of Brazilian individuals.

Methods:  This study employed a cross‐sectional design involving individuals from the State of Minas Gerais in the south‐eastern region of Brazil. Genomic DNA was obtained from oral swabs of 129 individuals and amplified using the polymerase chain reaction (PCR) with specific primers flanking the locus +3954 of IL1B. PCR products were submitted to restriction endonuclease digestion and analyzed by polyacrylamide gel electrophoresis, to distinguish alleles T and C of the IL1B gene, allowing for the determination of the genotypes and detection of the polymorphism.

Results and Conclusions:  The chronic periodontitis group displayed a higher percentage of the T allele (28%) when compared to the aggressive periodontitis group (10.7%, χ2 = 5.24, p = 0.02, OR = 0.31, CI = 0.11–0.88) and to control group (8.7%, χ2 = 7.11, p = 0.007, OR = 0.24, CI = 0.08–0.73). Our data suggested that the polymorphism in the locus +3954 of IL1B gene could be a risk factor for chronic periodontitis in a sample of Brazilian individuals.
Relationship between periodontal disease status and combination of biochemical assays of gingival crevicular fluid
Authors: Hanioka Takashi , Matsuse Ryoichi , Shigemoto Yukiko , Ojima Miki , Shizukuishi Satoshi
Published: June 2005

Background:  Currently, no biochemical assay involving gingival crevicular fluid is utilized routinely as a screening test for periodontal disease.

Objective:  The objective of the present study was to evaluate the potential of gingival crevicular fluid assay as a screening methodology.

Methods:  The subject population was comprised of 27 volunteers. Nine participants were classified as ‘subject with periodontal destruction’ (SPD) exhibiting at least one site with pocket depth and attachment loss > 3.5 mm, whereas the remaining individuals were categorized as ‘subject with minimal periodontal destruction’ (SMD). Gingival crevicular fluid was collected from fixed sites via a standardized method. Biochemical assays of 12 substances (hemoglobin, albumin, transferrin, α1‐antitrypsin, fibronectin, IgA, IgG, IgM, lactoferrin, myeloperoxidase and neutrophil elastase) were conducted at a commercial laboratory. Power transformation of total quantities in gingival crevicular fluid was performed for statistical analysis.

Results:  Relationships between total quantity of each substance and periodontal disease status were unclear. Logistic regression analysis yielded six predictive models, which consisted of substance pairs: neutrophil elastase/IgA, neutrophil elastase/hemoglobin, neutrophil elastase/α1‐antitrypsin and neutrophil elastase/IgG, and IgA/albumin and IgA/transferrin (p < 0.05). Regression lines for SPD and SMD on a scattergram of IgA and neutrophil elastase were nearly parallel within the range of amounts in gingival crevicular fluid. The predictive model derived from both substances afforded sensitivity and specificity of 88% and 94%, respectively.

Conclusions:  These results indicated that the combination of IgA and neutrophil elastase in gingival crevicular fluid may be crucial for prediction of periodontal disease status. Furthermore, these data suggested that biochemical assays employing both substances in gingival crevicular fluid may provide a satisfactory screening test for periodontal disease.
Periodontal status and preterm low birth weight: a case control study
Authors: Noack B. , Klingenberg J. , Weigelt J. , Hoffmann T.
Published: June 2005

Background:  Previous studies have suggested that periodontal disease may be an important risk factor for preterm low birth weight. However, the link between periodontal health status of pregnant women and preterm low birth weight is contentious, as recent studies found no association between periodontitis and pregnancy outcome.

Objective:  The aim of this study was to investigate this potential link in a German Caucasian population.

Methods:  Fifty‐nine pregnant women with a high risk for a preterm low birth weight infant (suffering from preterm contractions, cases, group 1) as well as 42 control women with no preterm contractions during pregnancy and having an infant appropriate for date and weight (≥ 37 weeks gestation, ≥ 2500 g, group 2) were examined. Clinical periodontal status was recorded on a full mouth basis. Subgingival plaque samples were taken and periodontal pathogens were identified by polymerase chain reaction. Additionally, interleukin‐1β level in gingival crevicular fluid was analysed.

Results:  The mean percentage of sites showing moderate to advanced attachment loss (≥ 3 mm) was low in all study groups (group 1: 9.9 ± 11.2%; group 2: 10.6 ± 14.1%, respectively). No significant differences between the groups in any aspects of the studied periodontitis parameters could be detected. Using a logistic regression model controlling for known preterm low birth weight risk factors, no periodontitis‐associated factors increased risk for preterm contractions or preterm low birth weight. The odds ratio (OR) was 1.19 for preterm contractions, the 95% confidence interval (CI) 0.46; 3.11 and 0.73 for preterm low birth weight; 95% CI: 0.13; 4.19, respectively.

Conclusion:  In this population, periodontitis was not a detectable risk factor for preterm low birth weight in pregnant women.
Inflammatory bone destruction and osteoimmunology
Authors: Takayanagi Hiroshi
Published: June 2005

Objectives:  The metabolism of hard tissue is influenced by the immune system. Research into the bone destruction associated with inflammatory diseases such as periodontal disease and rheumatoid arthritis has highlighted the importance of the interplay of the immune and skeletal systems. This interdisciplinary research field, called ‘osteoimmunology’, has become increasingly important for each system by itself as well as the biology linking them. The history and recent progress of this field are reviewed.

Material and methods:  ‘Osteoimmunology’ was coined to describe the pioneering work on the T‐cell regulation of osteoclastogenesis by the receptor activator of nuclear factor‐κB ligand (RANKL) and interferon (IFN)‐γ. Accumulating evidence suggests that the immune and skeletal systems share not only cytokines but also various signaling molecules, transcription factors and membrane receptors. The contribution of T cells to the pathogenesis of inflammatory bone destruction is discussed, and our recent findings are summarized to illustrate how the osteoimmunological network functions.

Results:  RANKL is an osteoclastogenic cytokine that links bone and the immune system. Immunomodulatory cytokines such as IFNs also participate in the regulation of RANKL signaling and inflammatory bone loss. The transcription factor nuclear factor of activated T cells c1 (NFATc1) has been identified as a master switch regulator of osteoclastogenesis. In addition, immunoglobulin‐like receptors are critically involved in bone homeostasis.

Conclusion:  Bone turns out to be a dynamic tissue that is constantly renewed, where the immune system participates to a hitherto unexpected extent. This emerging field will be of great importance to a better understanding and treatment of diseases of the skeletal and immune systems, as well as to the fundamental biology underpinning both.
Child and Adolescent Mental Health Services in the UK: Nurse Consultants
Authors: McDougall Tim
Published: June 2005

TOPIC:  Children's mental health services in the UK are transforming. Innovations in health, social welfare, education, and youth justice agencies have all got emotional literacy at their core. The changing face of children's services has called for partnerships and new ways of planning and delivering services in statutory and voluntary sectors. Alongside this, nursing is evolving and developing new ways to meet the challenges that modernization brings. Nurse consultants are children's champions who have a leading role in helping lead children's services into the 21st century.

PURPOSE:  The following article explores the role of nurse consultants in child and adolescent mental health services (CAMHS).
Microarray Analysis and Motif Detection Reveal New Targets of theSalmonella entericaSerovar Typhimurium HilA Regulatory Protein, IncludinghilAItself
Authors: De Keersmaecker Sigrid C. J. , Marchal Kathleen , Verhoeven Tine L. A. , Engelen Kristof , Vanderleyden Jos , Detweiler Corrella S.
Published: June 2005

DNA regulatory motifs reflect the direct transcriptional interactions between regulators and their target genes and contain important information regarding transcriptional networks. In silico motif detection strategies search for DNA patterns that are present more frequently in a set of related sequences than in a set of unrelated sequences. Related sequences could be genes that are coexpressed and are therefore expected to share similar conserved regulatory motifs. We identified coexpressed genes by carrying out microarray-based transcript profiling ofSalmonella entericaserovar Typhimurium in response to the spent culture supernatant of the probiotic strainLactobacillus rhamnosusGG. Probiotics are live microorganisms which, when administered in adequate amounts, confer a health benefit on the host. They are known to antagonize intestinal pathogens in vivo, including salmonellae.S. entericaserovar Typhimurium causes human gastroenteritis. Infection is initiated by entry of salmonellae into intestinal epithelial cells. The expression of invasion genes is tightly regulated by environmental conditions, as well as by many bacterial factors including the key regulator HilA. One mechanism by which probiotics may antagonize intestinal pathogens is by influencing invasion gene expression. Our microarray experiment yielded a cluster of coexpressedSalmonellagenes that are predicted to be down-regulated by spent culture supernatant. This cluster was enriched for genes known to be HilA dependent. In silico motif detection revealed a motif that overlaps the previously described HilA box in the promoter region of three of these genes,spi4_H,sicA, andhilA. Site-directed mutagenesis, β-galactosidase reporter assays, and gel mobility shift experiments indicated thatsicAexpression requires HilA and thathilAis negatively autoregulated.
The Genomic Island SGI1, Containing the Multiple Antibiotic Resistance Region ofSalmonella entericaSerovar Typhimurium DT104 or Variants of It, Is Widely Distributed in OtherS. entericaSerovars
Authors: Levings Renee S. , Lightfoot Diane , Partridge Sally R. , Hall Ruth M. , Djordjevic Steven P.
Published: June 2005

The global dissemination of the multiply-antibiotic-resistantSalmonella entericaserovar Typhimurium DT104 clone with the resistance genes located in a class 1 integron, here designated In104, within genomic island SGI1 is a significant public health issue. Here, we have shown that SGI1 and variants of it carrying different combinations of resistance genes are found in severalSalmonella entericaserovars. These are serovars Cerro, Derby, Dusseldorf, Infantis, Kiambu, and Paratyphi B dT+isolated from human infections and serovar Emek from sewage effluent. Two new variants, SGI1-I and SGI1-J, both of which include thedfrA1-orfC cassette array, were identified.
Functional Analysis of the
Burkholderia cenocepacia
ZmpA Metalloprotease
Authors: Kooi C. , Corbett C. R. , Sokol P. A.
Published: June 2005

Burkholderia cenocepacia
ZmpA is expressed as a preproenzyme typical of thermolysin-like proteases such as
Pseudomonas aeruginosa
LasB and
Bacillus thermoproteolyticus
thermolysin. The
zmpA
gene was expressed using the pPRO-EXHTa His
6
tag expression system, which incorporates a six-His tag at the N-terminal end of the protein, and recombinant ZmpA was purified using Ni-nitrilotriacetic acid affinity chromatography. Upon refolding of the recombinant His
6
-pre-pro-ZmpA (62 kDa), the fusion protein was autoproteolytically cleaved into 36-kDa (mature ZmpA) and 27-kDa peptides. Site-directed mutagenesis was employed to infer the identity of the active site residues of ZmpA and to confirm that the enzyme undergoes autoproteolytic cleavage. Oligonucleotide mutagenesis was used to replace H
465
with G
465
or A
465
, E
377
with A
377
or D
377
, or H
380
with P
380
or A
380
. Mutagenesis of H
465
, E
377
, or H
380
resulted in the loss of both autocatalytic activity and proteolytic activity. ZmpA with either substitution in H
380
was not detectable in
B. cenocepacia
cell extracts. The activity of the recombinant ZmpA was inhibited by EDTA and 1,10 phenanthroline, indicating that it is a zinc metalloprotease. ZmpA, however, was not inhibited by phosphoramidon, a classical inhibitor of the thermolysin-like proteases. The refolded mature ZmpA enzyme was proteolytically active against various substrates including hide powder azure, type IV collagen, fibronectin, neutrophil α-1 proteinase inhibitor, α
2
-macroglobulin, and gamma interferon, suggesting that
B. cenocepacia
ZmpA may cause direct tissue damage to the host or damage to host tissues through a modulation of the host's immune system.
Stable Accumulation of σ
54
in
Helicobacter pylori
Requires the Novel Protein HP0958
Authors: Pereira Lara , Hoover Timothy R.
Published: June 2005

Several flagellar genes in
Helicobacter pylori
are dependent on σ
54
(RpoN) for their expression. These genes encode components of the basal body, the hook protein, and a minor flagellin, FlaB. A protein-protein interaction map for
H. pylori
constructed from a high-throughput screen of a yeast two-hybrid assay (
http://pim.hybrigenics.com/pimrider
ext/common/) revealed interactions between σ
54
and the conserved hypothetical protein HP0958. To see if HP0958 influences σ
54
function, the corresponding gene was disrupted with a kanamycin resistance gene (
aphA3
) in
H. pylori
ATCC 43504 and the resulting mutant was analyzed. The
hp0958:aphA3
mutant was nonmotile and failed to produce flagella. Introduction of a functional copy of
hp0958
into the genome of the
hp0958:aphA3
mutant restored flagellar biogenesis and motility. The
hp0958:aphA3
mutant was deficient in expressing two σ
54
-dependent reporter genes,
flaB
′
-
′
xylE
and
hp1120
′
-
′
xylE
. Levels of σ
54
in the
hp0958
mutant were substantially lower than those in the parental strain, suggesting that the failure of the mutant to express the genes in the RpoN regulon and produce flagella was due to reduced σ
54
levels. Expressing σ
54
at high levels by putting
rpoN
under the control of the
ureA
promoter restored flagellar biogenesis and motility in the
hp0958:aphA3
mutant. Turnover of σ
54
was more rapid in the
hp0958:aphA3
mutant than it was in the wild-type strain, suggesting that HP0958 supports wild-type σ
54
levels in
H. pylori
by protecting it from proteolysis.
Staphylococcus aureus
ClpC Is Required for Stress Resistance, Aconitase Activity, Growth Recovery, and Death
Authors: Chatterjee Indranil , Becker Petra , Grundmeier Matthias , Bischoff Markus , Somerville Greg A. , Peters Georg , Sinha Bhanu , Harraghy Niamh , Proctor Richard A. , Herrmann Mathias
Published: June 2005

The ability of
Staphylococcus aureus
to adapt to various conditions of stress is the result of a complex regulatory response. Previously, it has been demonstrated that Clp homologues are important for a variety of stress conditions, and our laboratory has shown that a
clpC
homologue was highly expressed in the
S. aureus
strain DSM20231 during biofilm formation relative to expression in planktonic cells. Persistence and long-term survival are a hallmark of biofilm-associated staphylococcal infections, as cure frequently fails even in the presence of bactericidal antimicrobials. To determine the role of
clpC
in this context, we performed metabolic, gene expression, and long-term growth and survival analyses of DSM20231 as well as an isogenic
clpC
allelic-replacement mutant, a
sigB
mutant, and a
clpC sigB
double mutant. As expected, the
clpC
mutant showed increased sensitivity to oxidative and heat stresses. Unanticipated, however, was the reduced expression of the tricarboxylic acid (TCA) cycle gene
citB
(encoding aconitase), resulting in the loss of aconitase activity and preventing the catabolization of acetate during the stationary phase.
clpC
inactivation abolished post-stationary-phase recovery but also resulted in significantly enhanced stationary-phase survival compared to that of the wild-type strain. These data demonstrate the critical role of the ClpC ATPase in regulating the TCA cycle and implicate ClpC as being important for recovery from the stationary phase and also for entering the death phase. Understanding the stationary- and post-stationary-phase recovery in
S. aureus
may have important clinical implications, as little is known about the mechanisms of long-term persistence of chronic
S. aureus
infections associated with formation of biofilms.
Assimilation of Nicotinamide Mononucleotide Requires Periplasmic AphA Phosphatase inSalmonella enterica
Authors: Grose Julianne H. , Bergthorsson Ulfar , Xu Yaping , Sterneckert Jared , Khodaverdian Behzad , Roth John R.
Published: June 2005

Salmonella entericacan obtain pyridine from exogenous nicotinamide mononucleotide (NMN) by three routes. In route 1, nicotinamide is removed from NMN in the periplasm and enters the cell as the free base. In route 2, described here, phosphate is removed from NMN in the periplasm by acid phosphatase (AphA), and the produced nicotinamide ribonucleoside (NmR) enters the cell via the PnuC transporter. Internal NmR is then converted back to NMN by the NmR kinase activity of NadR. Route 3 is seen only inpnuC* transporter mutants, which import NMN intact and can therefore grow on lower levels of NMN. Internal NMN produced by either route 2 or route 3 is deamidated to nicotinic acid mononucleotide and converted to NAD by the biosynthetic enzymes NadD and NadE.
Transcriptional Analysis of the Conserved
ftsZ
Gene Cluster in
Mycoplasma genitalium
and
Mycoplasma pneumoniae
Authors: Benders Gwynedd A. , Powell Bradford C. , Hutchison Clyde A.
Published: June 2005

Several experimental approaches were used to construct a detailed transcriptional profile of the phylogenetically conserved
ftsZ
cell division gene cluster in both
Mycoplasma genitalium
and its closest relative,
Mycoplasma pneumoniae
. We determined initiation and termination points for the cluster, as well as an absolute steady-state RNA level for each gene. Transcription of this cluster in both these organisms was shown to be highly strand specific. While the four genes in this cluster are cotranscribed, their transcription unit also includes two genes of close proximity yet disparate function. A transcription initiation point immediately upstream of these two genes was detected in
M. genitalium
but not
M. pneumoniae
. In
M. pneumoniae
, transcription of the six genes terminates at a poly(U)-tailed hairpin. In
M. genitalium
, this transcription terminates at two closely spaced points by an unknown mechanism. Real-time reverse transcription-PCR analysis of this cluster in
M. pneumoniae
shows that mRNA levels for all six genes vary at most fivefold and form a gradient of decreasing quantity with increasing distance from the promoter at the beginning of the cluster. mRNA from coding regions was approximately 20- to 100-fold more abundant than that from intergenic regions. We estimated the most abundant mRNA we detected at 0.6 copy per cell. We conclude that groups of functionally related genes in
M. genitalium
and
M. pneumoniae
are often preceded by promoters but rarely followed by terminators. This causes functionally unrelated genes to be commonly cotranscribed in these organisms.
Epimerase Active Domain of
Pseudomonas aeruginosa
AlgG, a Protein That Contains a Right-Handed β-Helix
Authors: Douthit Stephanie A. , Dlakic Mensur , Ohman Dennis E. , Franklin Michael J.
Published: June 2005

The polysaccharide alginate forms a protective capsule for
Pseudomonas aeruginosa
during chronic pulmonary infections. The structure of alginate, a linear polymer of β1-4-linked O-acetylated
d
-mannuronate (M) and
l
-guluronate (G), is important for its activity as a virulence factor. Alginate structure is mediated by AlgG, a periplasmic C-5 mannuronan epimerase. AlgG also plays a role in protecting alginate from degradation by the periplasmic alginate lyase AlgL. Here, we show that the C-terminal region of AlgG contains a right-handed β-helix (RHβH) fold, characteristic of proteins with the
c
arbohydrate-binding
a
nd
s
ugar
h
ydrolase (CASH) domain. When modeled based on pectate lyase C of
Erwinia chrysanthemi
, the RHβH of AlgG has a long shallow groove that may accommodate alginate, similar to protein/polysaccharide interactions of other CASH domain proteins. The shallow groove contains a 324-DPHD motif that is conserved among AlgG and the extracellular mannuronan epimerases of
Azotobacter vinelandii
. Point mutations in this motif disrupt mannuronan epimerase activity but have no effect on alginate secretion. The D324A mutation has a dominant negative phenotype, suggesting that the shallow groove in AlgG contains the catalytic face for epimerization. Other conserved motifs of the epimerases, 361-NNRSYEN and 381-NLVAYN, are predicted to lie on the opposite side of the RHβH from the catalytic center. Point mutations N362A, N367A, and V383A result in proteins that do not protect alginate from AlgL, suggesting that these mutant proteins are not properly folded or not inserted into the alginate biosynthetic scaffold. These motifs are likely involved in asparagine and hydrophobic stacking, required for structural integrity of RHβH proteins, rather than for mannuronan catalysis. The results suggest that the AlgG RHβH protects alginate from degradation by AlgL by channeling the alginate polymer through the proposed alginate biosynthetic scaffold while epimerizing approximately every second
d
-mannuronate residue to
l
-guluronate along the epimerase catalytic face.
Identification of a Second Collagen-Like Glycoprotein Produced by
Bacillus anthracis
and Demonstration of Associated Spore-Specific Sugars
Authors: Waller Lashanda N. , Stump Michael J. , Fox Karen F. , Harley William M. , Fox Alvin , Stewart George C. , Shahgholi Mona
Published: June 2005

Certain carbohydrates (rhamnose, 3-
O
-methyl rhamnose, and galactosamine) have been demonstrated to be present in
Bacillus anthracis
spores but absent in vegetative cells. Others have demonstrated that these spore-specific sugars are constituents of the glycoprotein BclA. In the current work, spore extracts were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A second collagen-like glycoprotein, BclB, was identified in
B. anthracis
. The protein moiety of this glycoprotein was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MS) and the carbohydrate components by gas chromatography-mass spectrometry and tandem mass spectrometry. Spore-specific sugars were also demonstrated to be components of BclB.
Genomic Sequence of an Otitis Media Isolate of Nontypeable
Haemophilus influenzae
: Comparative Study with
H. influenzae
Serotype d, Strain KW20
Authors: Harrison Alistair , Dyer David W. , Gillaspy Allison , Ray William C. , Mungur Rachna , Carson Matthew B. , Zhong Huachun , Gipson Jenny , Gipson Mandy , Johnson Linda S. , Lewis Lisa , Bakaletz Lauren O. , Munson Robert S.
Published: June 2005

In 1995, the Institute for Genomic Research completed the genome sequence of a rough derivative of
Haemophilus influenzae
serotype d, strain KW20. Although extremely useful in understanding the basic biology of
H. influenzae
, these data have not provided significant insight into disease caused by nontypeable
H. influenzae
, as serotype d strains are not pathogens. In contrast, strains of nontypeable
H. influenzae
are the primary pathogens of chronic and recurrent otitis media in children. In addition, these organisms have an important role in acute otitis media in children as well as other respiratory diseases. Such strains must therefore contain a gene repertoire that differs from that of strain Rd. Elucidation of the differences between these genomes will thus provide insight into the pathogenic mechanisms of nontypeable
H. influenzae
. The genome of a representative nontypeable
H. influenzae
strain, 86-028NP, isolated from a patient with chronic otitis media was therefore sequenced and annotated. Despite large regions of synteny with the strain Rd genome, there are large rearrangements in strain 86-028NP's genome architecture relative to the strain Rd genome. A genomic island similar to an island originally identified in
H. influenzae
type b is present in the strain 86-028NP genome, while the mu-like phage present in the strain Rd genome is absent from the strain 86-028NP genome. Two hundred eighty open reading frames were identified in the strain 86-028NP genome that were absent from the strain Rd genome. These data provide new insight that complements and extends the ongoing analysis of nontypeable
H. influenzae
virulence determinants.
Architecture and Adhesive Activity of the
Haemophilus influenzae
Hsf Adhesin
Authors: Cotter Shane E. , Yeo Hye-Jeong , Juehne Twyla , St. Geme Joseph W.
Published: June 2005

Haemophilus influenzae
type b is an important cause of meningitis and other serious invasive diseases and initiates infection by colonizing the upper respiratory tract. Among the major adhesins in
H. influenzae
type b is a nonpilus protein called Hsf, a large protein that forms fiber-like structures on the bacterial surface and shares significant sequence similarity with the nontypeable
H. influenzae
Hia autotransporter. In the present study, we characterized the structure and adhesive activity of Hsf. Analysis of the predicted amino acid sequence of Hsf revealed three regions with high-level homology to the HiaBD1 and HiaBD2 binding domains in Hia. Based on examination of glutathione
S
-transferase fusion proteins corresponding to these regions, two of the three had adhesive activity and one was nonadhesive in assays with cultured epithelial cells. Structural modeling demonstrated that only the two regions with adhesive activity harbored an acidic binding pocket like the binding pocket identified in the crystal structure of HiaBD1. Consistent with these results, disruption of the acidic binding pockets in the adhesive regions eliminated adhesive activity. These studies advance our understanding of the architecture of Hsf and the family of trimeric autotransporters and provide insight into the structural determinants of
H. influenzae
type b adherence.