bluuebunny/crossref_metadata_2025 huggingface dataset:
Peptide alarmone signalling triggers an auto‐active bacteriocin necessary for genetic competence
Authors: Perry Julie A. , Jones Marcus B. , Peterson Scott N. , Cvitkovitch Dennis G. , Lévesque Céline M.
Published: April 2009

The induction of genetic competence is a strategy used by bacteria to increase their genetic repertoire under stressful environmental conditions. Recently, Streptococcus pneumoniae has been shown to co‐ordinate the uptake of transforming DNA with fratricide via increased expression of the peptide pheromone responsible for competence induction. Here, we document that environmental stress‐induced expression of the peptide pheromone competence‐stimulating peptide (CSP) in the oral pathogen Streptococcus mutans. We showed that CSP is involved in the stress response and determined the CSP‐induced regulon in S. mutans by microarray analysis. Contrary to pneumococcus, S. mutans responds to increased concentrations of CSP by cell lysis in only a fraction of the population. We have focused on the mechanism of cell lysis and have identified a novel bacteriocin as the ‘death effector’. Most importantly, we showed that this bacteriocin causes cell death via a novel mechanism of action: intracellular action against self. We have also identified the cognate bacteriocin immunity protein, which resides in a separate unlinked genetic locus to allow its differential regulation. The role of the lytic response in S. mutans competence is also discussed. Together, these findings reveal a novel autolytic pathway in S. mutans which may be involved in the dissemination of fitness‐enhancing genes in the oral biofilm.
Ectopic production of VapCs from Enterobacteria inhibits translation and trans‐activates YoeB mRNA interferase
Authors: Winther Kristoffer S. , Gerdes Kenn
Published: April 2009

Toxin–antitoxin loci have been identified in almost all free‐living prokaryotes, often in high copy numbers. The biological function and molecular targets of the abundant vapBC loci are not yet known. Here we analyse the vapBC loci of Salmonella LT2 and Shigella plasmid pMYSH6000. Both loci encode putative PIN (PilT N‐terminal) domain toxins, and antitoxins that may regulate vapBC transcription. We show that vapBCLT2 and vapBCpMYSH are bona fide TA loci: (i) both VapCs inhibited cell growth very efficiently and were counteracted by the cognate VapBs; (ii) both VapCs inhibited translation; (iii) transcription of the vapBC loci was induced by amino acid starvation and chloramphenicol, consistent with the proposal that VapB is an unstable inhibitor of vapBC transcription; (iv) ectopic expression of both VapCs induced a bacteriostatic condition that could be reversed by the cognate antitoxins. Unexpectedly, induction of vapC in Escherichia coli resulted in mRNA cleavage at stop‐codons. Surprisingly, these cleavages depended on the yefM yoeB locus, indicating cross‐activation between different toxins, that is, VapC activated YoeB mRNA interferase. Activation of YoeB depended on Lon, indicating that Lon degrades YefM antitoxin. Based on these results we present a model that explains activation of YoeB.
Biosynthesis of the red pigment bikaverin in Fusarium fujikuroi: genes, their function and regulation
Authors: Wiemann Philipp , Willmann Anita , Straeten Marcus , Kleigrewe Karin , Beyer Marita , Humpf Hans‐Ulrich , Tudzynski Bettina
Published: April 2009

Fusarium secondary metabolites are structurally diverse, have a variety of activities and are generally poorly understood biosynthetically. The F. fujikuroi polyketide synthase gene bik1 was previously shown to be responsible for formation of the mycelial pigment bikaverin. Here we present the characterization of five genes adjacent to bik1 as encoding a putative FAD‐dependent monooxygenase (bik2), an O‐methyltransferase (bik3), an NmrA‐like protein (bik4), a Zn(II)2Cys6 transcription factor (bik5) and an MFS transporter (bik6). Deletion of each gene resulted in total loss or significant reduction of bikaverin synthesis. Expression studies revealed that all bik genes are repressed by high amounts of nitrogen in an AreA‐independent manner and are subject to a time‐ and pH‐dependent regulation. Deletion of the pH regulatory gene pacC resulted in partial derepression while complementation with a dominant active allele resulted in repression of bik genes at acidic ambient pH. Transcription of all bik genes in strains lacking bik1, bik2 or bik3 was essentially eliminated, while transcription of some bik genes was detected in strains lacking bik4, bik5 or bik6. Thus, bikaverin synthesis is regulated by a complex regulatory network. Understanding how different factors influence the synthesis of this model secondary metabolite will aid understanding secondary metabolism in general.
Characterization of staphyloferrin A biosynthetic and transport mutants in Staphylococcus aureus
Authors: Beasley Federico C. , Vinés Enrique D. , Grigg Jason C. , Zheng Qin , Liu Suya , Lajoie Gilles A. , Murphy Michael E. P. , Heinrichs David E.
Published: April 2009

Iron is critical for virtually all forms of life. The production of high‐affinity iron chelators, siderophores, and the subsequent uptake of iron–siderophore complexes are a common strategy employed by microorganisms to acquire iron. Staphylococcus aureus produces siderophores but genetic information underlying their synthesis and transport is limited. Previous work implicated the sbn operon in siderophore synthesis and the sirABC operon in uptake. Here we characterize a second siderophore biosynthetic locus in S. aureus; the locus consists of four genes (in strain Newman these open reading frames are designated NWMN_2079–2082) which, together, are responsible for the synthesis and export of staphyloferrin A, a polycarboxylate siderophore. While deletion of the NWMN_2079–2082 locus did not affect iron‐restricted growth of S. aureus, strains bearing combined sbn and NWMN_2079–2082 locus deletions produced no detectable siderophore and demonstrated severely attenuated iron‐restricted growth. Adjacent to NWMN_2079–2082 resides the htsABC operon, encoding an ABC transporter previously implicated in haem acquisition. We provide evidence here that HtsABC, along with the FhuC ATPase, is required for the uptake of staphyloferrin A. The crystal structure of apo‐HtsA was determined and identified a large positively charged region in the substrate‐binding pocket, in agreement with a role in binding of anionic staphyloferrin A.
Impaired TLR‐2 expression and TLR‐2‐mediated cytokine secretion in macrophages from patients with atopic dermatitis
Authors: Niebuhr M. , Lutat C. , Sigel S. , Werfel T.
Published: April 2009

Background:  In many patients with atopic dermatitis (AD), the disease is complicated by their enhanced susceptibility to bacterial skin infections, especially with Staphylococcus aureus. The pattern recognition receptor toll‐like receptor (TLR)‐2 recognizes components of S. aureus, for example, lipoteichoic acid (LTA) and peptidoglycan (PGN) and, therefore, might be crucial in the pathogenesis and flare‐ups of AD.

Objective:  To investigate TLR‐2 expression and cytokine secretion in macrophages from patients with AD compared to healthy controls upon TLR‐2 stimulation with PGN, LTA and Pam3Cys.

Methods:  Macrophages were cultivated from highly purified peripheral blood monocytes of AD patients and nonatopic healthy controls and stimulated with PGN, LTA and Pam3Cys in a time and dose–dependent manner. Afterwards, TLR‐2 expression and cytokine secretion were measured on protein and mRNA level. TLR‐1 and TLR‐6 expression were investigated on the mRNA level. Immunohistochemical stainings from punch biopsies were performed to investigate TLR‐2 expression in skin macrophages.

Results:  We could clearly show that macrophages from patients with AD expressed significantly less TLR‐2, whereas the expression pattern of TLR‐1 and TLR‐6 were not altered. Macrophages had a reduced capacity to produce pro‐inflammatory cytokines such as IL‐6, IL‐8 and IL‐1β after stimulation with TLR‐2 ligands.

Conclusion:  Our findings clearly show an impaired TLR‐2 expression and functional differences of TLR‐2‐mediated effects on macrophages of AD patients compared to healthy controls which might contribute to the enhanced susceptibility to skin infections with S. aureus in AD.
The size of the disease relevant IgE antibody fraction in relation to ‘total‐IgE’ predicts the efficacy of anti‐IgE (Xolair®) treatment
Authors: Johansson S. G. O. , Nopp A. , Öman H. , Ankerst J. , Cardell L. O. , Grönneberg R. , Matsols H. , Rudblad S. , Strand V. , Stålenheim G.
Published: April 2009

Background:  Some patients with allergic asthma treated with anti‐IgE (Xolair®) do not become symptom free. Better criteria for response assessment than allergy skin tests or IgE determination are needed. The impact of the size of the disease relevant allergen‐specific IgE antibody fraction, i.e. the percentage of IgE antibody of total IgE, was evaluated in cat allergic patients treated with the recommended doses of Xolair®. Results were measured as changes in basophil allergen threshold sensitivity (CD‐sens).

Methods:  In a double‐blind placebo controlled trial 20 patients with a high (>3.8%) and 18 with a low (<1%) percentage of IgE antibodies to cat were given Xolair® for 16 weeks and the change in CD‐sens was compared to 11 and 10 patients, respectively, in each group receiving placebo.

Results:  The CD‐sens dropped significantly in both the high (P < 0.001) and low (P < 0.001) group on Xolair® but did not change significantly after placebo. For Xolair®‐treated patients, at the end of the trial there was a highly significant (P < 0.001) difference in CD‐sens between the high group, where no patients, and the low group, where 13/18 patients, had become negative.

Conclusions:  The currently recommended doses of Xolair® very efficiently eliminate IgE antibodies if the IgE antibody fraction is <1% of total IgE but has not enough effect on allergen sensitivity if the fraction is >3–4%. Further studies will show if increased doses of Xolair® would help also these patients, who seem to represent about 1/3 of the patient population.
Genetic variation in CRTh2 influences development of allergic phenotypes
Authors: Cameron L. , Depner M. , Kormann M. , Klopp N. , Illig T. , Von Mutius E. , Kabesch M.
Published: April 2009

Background:  Allergic disorders are characterized by an increase in the Th2 cytokines IL‐4, IL‐5 and IL‐13, produced primarily by Th2 cells. These cells are marked by the expression of CRTh2 (chemoattractant receptor‐homologous molecule expressed on Th2 cells), a receptor for prostaglandin D2. As genetic variation plays a significant role in the predisposition for allergic disorders, we investigated the influence of single nucleotide polymorphisms (SNPs) in CRTh2.

Methods:  In a large study population of German children (n = 4264) from the International Study of Asthma and Allergy in Children (ISAAC II), six polymorphisms in CRTh2 were genotyped. Statistical analyses were performed using single SNP and haplotype analyses.

Results:  Uncorrected associations among −6373G>A, +1431G>C and +1538A>G were observed with a number of allergic phenotypes (P < 0.05). After correction, association between +1431C and specific IgE to food allergens remained significant (P = 0.04). Associations of haplotype (H)3 (containing +1538G) with reduced risk for asthma and H2 (containing +1431C) with increased risk for specific IgE to food allergens also remained significant after correction for multiple testing (P = 0.004).

Conclusions:  Genetic variation within CRTh2 modifies the development of allergic sensitization and asthma in a population of German children.
Decreased FOXP3 protein expression in patients with asthma
Authors: Provoost S. , Maes T. , Van Durme Y. M. , Gevaert P. , Bachert C. , Schmidt‐Weber C. B. , Brusselle G. G. , Joos G. F. , Tournoy K. G.
Published: April 2009

Background:  T‐regulatory cells (Treg) are important in balancing immune responses and maintaining peripheral tolerance. Current evidence suggests that asthma is characterized by a relative deficiency in Treg, allowing T helper 2 cells to expand. In this study, we aimed to evaluate circulating Treg, defined by the protein FOXP3, in both control subjects and patients with stable asthma.

Methods:  Peripheral blood mononuclear cells (PBMC) of control (n = 14) and asthmatic patients (n = 29) were labeled for CD4, CD25, and intracellular FOXP3 and analyzed using flow cytometry. In CD3/CD28 stimulated PBMC, the effects of dexamethasone on the transcription factors T‐bet, GATA‐3, FOXP3, and RORc2 and representative cytokines were studied.

Results:  In control subjects and asthmatic patients, numbers of peripheral blood CD4+CD25high and CD4+CD25highFOXP3+ T‐cells were similar. However, FOXP3 protein expression within CD4+CD25high T‐cells was significantly decreased in asthmatic patients. There was a tendency for increased FOXP3 expression within CD4+CD25high T‐cells in glucocorticosteroid‐treated patients when compared to steroid‐naive asthmatic patients. In stimulated PBMC, dexamethasone treatment increased the anti‐/proinflammatory transcription ratios of FOXP3/GATA‐3, FOXP3/T‐bet, and FOXP3/RORc2.

Conclusion:  Asthmatic patients have decreased FOXP3 protein expression within their CD4+CD25high Treg. Our findings also suggest that treatment with inhaled glucocorticosteroids in asthmatics might increase this FOXP3 protein expression within the CD4+CD25high T‐cell population.
Neutrophilic airway inflammation is a main feature of induced sputum in nonatopic asthmatic children
Authors: Drews A. C. , Pizzichini M. M. M. , Pizzichini E. , Pereira M. U. , Pitrez P. M. , Jones M. H. , Sly P. D. , Stein R. T.
Published: April 2009

Background:  Asthma phenotypes are well described among children. However, there are few studies comparing airway inflammation in different clinical presentations of pediatric asthma. We tested the hypothesis that nonatopic asthma is associated with a predominant noneosinophilic inflammation in the airways, as assessed by induced sputum. The objective of this study was to evaluate the cytological characteristics of induced sputum (IS) in atopic (AA), nonatopic asthmatics (NAA) and nonatopic nonasthmatic children (NANA).

Methods:  Of 90 selected children, 77 met eligibility criteria for performing IS and were classified as: AA, n = 28, NAA, n = 29 and NANA, n = 19. Subjects answered to a set of ISAAC‐based questions and were skin‐tested for common aeroallergens. A defined series of exclusion criteria was applied.

Results:  Induced sputum was obtained from 54 (70.1%) subjects (21 AA, 20 NAA and 13 NANA). Demographic data and mean FEV1 were similar in the three groups. The proportion of eosinophils [median, inter quartile range (IQR)] was significantly higher in the sputum of AA [(6.0.)12)] compared with NAAs [0 (2)] and NANAs [0 (1)], P < 0.001. The proportion of children with sputum eosinophilia (eos > 3%) was also significantly higher in AA (71.4%) when compared with NAA (28.6%); none of the NANA had sputum eosinophilia. Nonatopic asthmatic children had significantly higher proportions and absolute number of neutrophils than AA and controls.

Conclusions:  The results suggest that nonatopic children present IS with a cell pattern that is predominantly neutrophilic while eosinophilia is the hallmark of airway inflammation in the majority of atopic wheezing children not treated with inhaled steroids.
Cross‐reactivity and tolerability of imipenem in patients with delayed‐type, cell‐mediated hypersensitivity to β‐lactams
Authors: Schiavino D. , Nucera E. , Lombardo C. , Decinti M. , Pascolini L. , Altomonte G. , Buonomo A. , Patriarca G.
Published: April 2009

Background: Administration of imipenem‐cilastatin to patients with IgE‐mediated hypersensitivity to β‐lactams has always been considered potentially harmful. Recent studies have demonstrated the tolerability of carbapenems (imipenem‐cilastatin and meropenem) in patients with IgE‐mediated hypersensitivity to β‐lactams; there are no studies on this topic regarding patients with cell‐mediated allergy to β‐lactams. The aim of this study is to assess cross‐reactivity and tolerability of imipenem in patients with cell‐mediated allergy to β‐lactams.

Methods: From our database we selected 73 patients with cell‐mediated allergy to β‐lactams, diagnosed by means of immediate‐type skin tests, delayed reading intradermal tests, patch tests and detection of specific IgE. Patients with negative patch tests with imipenem‐cilastatin underwent an intramuscular test dosing.

Results: Our patients had a total of 94 nonimmediate reactions to penicillins. All patients had positive patch tests and/or delayed reading intradermal tests for at least one of the penicillin reagent tested and negative immediate‐type skin tests and specific IgE. Four patients out of 73 had a positive patch tests to at least one penicillin reagent and imipenem‐cilastatin showing cross‐reactivity. Sixty‐four patients underwent the imipenem‐cilastatin intramuscular test dosing and none of them had a clinical reaction.

Conclusions: Our rate of cross‐reactivity between imipenem‐cilastatin and other β‐lactams was 5.5%. This result is different from previous findings and this may be explained by the fact that we investigated patients with cell‐mediated allergy to β‐lactams. Patients with cell‐mediated allergy to β‐lactams should undergo patch tests and a tolerance challenge test before treatment with imipenem‐cilastatin.
Oral cavity is not a reservoir for Helicobacter pylori in infected patients with functional dyspepsia
Authors: Silva Rossi‐Aguiar V. P. , Navarro‐Rodriguez T. , Mattar R. , Siqueira de Melo Peres M. P. , Correa Barbuti R. , Silva F. M. , Carrilho F. J. , Eisig J. N.
Published: April 2009

Introduction:  Helicobacter pylori infection is very prevalent in Brazil, infecting almost 65% of the population. The aim of this study was to evaluate the presence of this bacterium in the oral cavity of patients with functional dyspepsia (epigastric pain syndrome), establish the main sites of infection in the mouth, and assess the frequency of cagA and vacA genotypes of oral H. pylori.

Methods:  All 43 outpatients with epigastric pain syndrome, who entered the study, were submitted to upper gastrointestinal endoscopy to rule out organic diseases. Helicobacter pylori infection in the stomach was confirmed by a rapid urease test and urea breath tests. Samples of saliva, the tongue dorsum and supragingival dental plaque were collected from the oral cavity of each subject and subgingival dental plaque samples were collected from the patients with periodontitis; H. pylori infection was verified by polymerase chain reaction using primers that amplify the DNA sequence of a species‐specific antigen present in all H. pylori strains; primers that amplify a region of urease gene, and primers for cagA and vacA (m1, m2, s1a, s1b, s2) genotyping.

Results:  Thirty patients harbored H. pylori in the stomach, but it was not possible to detect H. pylori in any oral samples using P1/P2 and Urease A/B. The genotype cagA was also negative in all samples and vacA genotype could not be characterized (s‐m‐).

Conclusion:  The oral cavity may not be a reservoir for H. pylori in patients with epigastric pain syndrome, the bacterium being detected exclusively in the stomach.
Prevalence, intensity and identity of bacteraemia following conservative dental procedures in children
Authors: Sonbol H. , Spratt D. , Roberts G. J. , Lucas V. S.
Published: April 2009

Introduction:  The aim of this study was to investigate the prevalence, intensity and microbial identity of bacteraemia following conservative dental procedures. The procedures were placement of rubber dam, use of the fast drill, use of the slow drill and placement of a matrix band and wedge.

Method:  Two hundred and five children and adolescents undergoing general anaesthesia for dental treatment at the Eastman Dental Hospital were recruited. Each subject was randomly allocated to one of the procedure groups. A baseline blood sample was taken before any dental treatment was carried out. A second blood sample was taken 30 s after a single conservative procedure. The blood samples were processed using lysis filtration. All bacterial isolates were identified using comparative 16 S ribosomal RNA gene sequencing. Oral Streptococcus spp. and coagulase‐negative Staphylococcus spp. were further identified by comparative sodA gene sequencing.

Results:  The prevalence of bacteraemia was significantly greater following placement of rubber dam (P = 0.01) and placement of matrix band and wedge, compared with baseline. The intensity of bacteraemia was significantly greater following placement of rubber dam (P = 0.001) and placement of matrix band and wedge (P = 0.0001). The most frequently isolated bacteria were Streptococcus spp. (56%), Actinomyces spp. (15%) and coagulase‐negative Staphylococcus spp. (15%).

Conclusion:  Conservative dental procedures are a significant cause of bacteraemia.
Expression of receptor activator of nuclear factor‐κB ligand by B cells in response to oral bacteria
Authors: Han X. , Lin X. , Seliger A. R. , Eastcott J. , Kawai T. , Taubman M. A.
Published: April 2009

Introduction:  We investigated receptor activator of nuclear factor‐κB ligand (RANKL) expression by B lymphocytes during early and late aspects of the immune response to Aggregatibacter actinomycetemcomitans, a gram‐negative, anaerobic bacterium associated with aggressive periodontal disease.

Methods:  Expression of messenger RNA transcripts (tumor necrosis factor‐α, Toll‐like receptors 4 and 9, interleukins 4 and 10, and RANKL) involved in early (1‐day) and late (10‐day) responses in cultured rat splenocytes was examined by reverse transcription–polymerase chain reaction (RT‐PCR). The immune cell distribution (T, B, and natural killer cells and macrophages) in cultured rat splenocytes and RANKL expression in B cells were determined by flow cytometric analyses. B‐cell capacity for induction of osteoclast differentiation was evaluated by coculture with RAW 264.7 cells followed by a tartrate‐resistant acid phosphatase (TRAP) activity assay.

Results:  The expression levels of interleukins 4 and 10 in cultured cells were not changed in the presence of A. actinomycetemcomitans until cultured for 3 days, and peaked after 7 days. After culture for 10 days, the percentages of B and T cells, the overall RANKL messenger RNA transcripts, and the percentage of RANKL‐expressing immunoglobulin G‐positive cells were significantly increased in the presence of A. actinomycetemcomitans. These increases were considerably greater in cells isolated from A. actinomycetemcomitans‐immunized animals than from non‐immunized animals. RAW 264.7 cells demonstrated significantly increased TRAP activity when cocultured with B cells from A. actinomycetemcomitans‐immunized animals. The addition of human osteoprotegerin‐Fc to the culture significantly diminished such increases.

Conclusion:  This study suggests that B‐lymphocyte involvement in the immune response to A. actinomycetemcomitans through upregulation of RANKL expression potentially contribute to bone resorption in periodontal disease.
Comparative whole‐genome analysis of Streptococcus mutans isolates within and among individuals of different caries status
Authors: Zhang L. , Foxman B. , Drake D. R. , Srinivasan U. , Henderson J. , Olson B. , Marrs C. F. , Warren J. J. , Marazita M. L.
Published: April 2009

Introduction:  Genotypic analyses of Streptococcus mutans using fingerprinting methods depend on a few genetic loci being different but do not reveal the underlying genome‐wide differences between strains.

Methods:  We used comparative genomic hybridization (CGH) with 70‐mer oligonucleotide microarrays containing open reading frames (ORFs) from S. mutans strain UA159 to examine the genetic diversity of 44 isolates from nine children selected from a local study population in Eastern Iowa.

Results:  Unique strains (clones) within each child initially identified by arbitrary‐priming polymerase chain reaction were confirmed by CGH. There was a wide range of variation in the hybridization patterns of the 1948 ORFs among the test isolates examined. Between 87 and 237 ORFs failed to give a positive signal among individual isolates. A total of 323 of the UA159 ORFs were absent from one or more of the test strains. These 323 variable genes seemed to be distributed across the entire UA159 genome and across all the predicted functional categories.

Conclusion:  This set of very close geographically and temporally collected S. mutans isolates had a degree of gene content variation as high as a previously examined global set of strains. Comparing the frequency of these variable genes, the majority of which have unknown function, among strains of different origins (i.e. different caries status) could help to determine their relevance in S. mutans cariogenicity.
Genes responsible for dextran‐dependent aggregation of Streptococcus sobrinus strain 6715
Authors: Sato Y. , Okamoto‐Shibayama K. , Takada K. , Igarashi T. , Hirasawa M.
Published: April 2009

Introduction:  Streptococcus sobrinus exhibits more significant dextran‐dependent aggregation mediated by glucan‐binding proteins than Streptococcus mutans. We have identified four glucan‐binding protein C gene (gbpC) homologues designated as gbpC1, gbpC2, dblA and dblB in S. sobrinus in contrast to the single gene gbpC in S. mutans. We attempted to determine which gene is most responsible for the dextran‐dependent aggregation of S. sobrinus.

Methods:  We introduced mutation with a chemical mutagen, 1‐methyl‐3‐nitro‐1‐nitrosoguanidine, into S. sobrinus strain 6715 and analysed the four gbpC homologous gene sequences in the parental strain 6715 and an obtained aggregation‐negative mutant NUM‐Ssg99. We also examined the localization of proteins encoded by these genes in the mutant NUM‐Ssg99.

Results:  The nucleotide sequences of the gbpC1, gbpC2 and dblA genes in NUM‐Ssg99 were 100% identical to the homologous genes in parental strain 6715. In contrast, a truncated mutation was detected in the dblB gene and the mutant protein devoid of the LPXTG motif was confirmed by Western blot analysis to be released into the extracellular milieu.

Conclusion:  We conclude that the dblB gene among the four GbpC homologous protein genes is most responsible for aggregation in strain 6715.
Efficiency of oral fluid collection devices in extracting antibodies
Authors: Chang C. K. , Cohen M. E. , Bienek D. R.
Published: April 2009

Introduction:  To facilitate diagnoses, this study determined the efficacy of commercial oral fluid collection devices for their ability to recover three human immunoglobulin isotypes; immunoglobulin A (IgA), IgG, and IgM.

Methods:  The sandwich enzyme‐linked immunosorbent assay was used to determine antibody recovery from the following devices: (i) OraSure® oral specimen collection device, (ii) saliva•sampler®, (iii) ORALscreen™ collector, (iv) Dri‐Angle®, (v) no. 2 cotton roll, (vi) all‐gauze sponges device, and (vii) DentaSwabs®. For each isotype tested, the recovered eluate was compared with the concentration applied to the device. The performance of each device was determined at various antibody concentrations.

Results:  Recovery of IgA from the saliva•sampler, ORALscreen collector, Dri‐Angle and cotton roll was comparable to that seeded onto the device. When compared with the seeded IgG concentration, the mean concentration of antibody recovered by each product differed by approximately ± 9 ng/ml. The average amount of IgM recovered by the cotton roll and all‐gauze sponges device was approximately 29 and 39 ng/ml, respectively, less (P < 0.0001) than that seeded on the device. For all isotypes tested, the amount of antibody recovered from the device was dependent on the initial seeding concentration.

Conclusion:  Collectively, these data suggest that the product used for specimen collection can affect retrieval of antibodies and potentially confound patient diagnosis.
Human cytomegalovirus and Epstein–Barr virus inhibit oral bacteria‐induced macrophage activation and phagocytosis
Authors: Lin Y.‐L. , Li M.
Published: April 2009

Introduction:  Periodontal disease is an inflammatory condition caused by periodontal microorganisms. Viruses such as human cytomegalovirus (HCMV) and Epstein–Barr virus (EBV) are associated with certain types of periodontal disease, but their roles in promoting the disease are unclear. Because both viruses infect human macrophages, cells which play key roles in the clearance of pathogenic bacteria, it is likely that the viruses alter the functional capacity of macrophages by inhibiting their defense mechanisms against invading pathogens.

Methods:  Macrophages preinfected with HCMV or EBV were evaluated following stimulation by selected oral bacteria. Bacteria‐induced macrophage activation was assayed by measuring the levels of tumor necrosis factor‐α (TNF‐α) produced in the media, and phagocytic activity was analysed by a phagocytosis assay with fluorescein isothiocyanate‐labeled bacteria. The virus‐infected macrophages were also subjected to semi‐quantitative polymerase chain reaction to measure the expression of toll‐like receptor 9, which is involved in the activation of phagocytosis‐related pathways.

Results:  Both HCMV and EBV significantly diminished the TNF‐α production typically induced by oral bacteria, inhibited the phagocytic activity of macrophages, and downregulated the expression of toll‐like receptor 9.

Conclusion:  Infection by HCMV or EBV inhibits the functional ability of macrophages to respond to bacterial challenge, thereby suggesting their pathogenic role in the development of periodontal disease.
Oral Candida infection and colonization in solid organ transplant recipients
Authors: Dongari‐Bagtzoglou A. , Dwivedi P. , Ioannidou E. , Shaqman M. , Hull D. , Burleson J.
Published: April 2009

Introduction:  Oral Candida carriage and infection have been reported to be associated with a greater risk for systemic infection in transplant recipients; however, a systematic analysis of the oral Candida titers and species has not been previously conducted. The objectives of this study were to determine the prevalence of oropharyngeal candidiasis, the oral carrier status, Candida titers and species in this population.

Methods:  Ninety kidney and heart transplant subjects and 72 age‐matched healthy controls were included. Swabs from the oral mucosa and a standardized amount of unstimulated saliva were plated on Chromagar™Candida, and colony‐forming units per millilitre were calculated. Initial speciation was based on colony color and was confirmed by standard germ tube, biotyping, or polymerase chain reaction assays.

Results:  Infection with C. albicans was detected in seven transplant subjects and none of the controls. The transplant group had significantly higher oral Candida titers than the control group. There were no statistically significant relationships between the dose or type of immunosuppressants and oral Candida titers or infection. A significantly higher percentage of transplant subjects were colonized by more than one species, compared with control subjects. The most frequent species combination in transplant subjects was C. albicans and C. glabrata. C. glabrata was isolated from 13.5% of transplant carriers and none of the controls.

Conclusions:  Increased oral Candida infection and carriage titers were found in the transplant population. Although the majority of transplant patients were colonized by C. albicans, C. glabrata appears to emerge as the second most prevalent species.
Homotypic biofilm structure of Porphyromonas gingivalis is affected by FimA type variations
Authors: Kuboniwa M. , Amano A. , Inaba H. , Hashino E. , Shizukuishi S.
Published: April 2009

Introduction:  Porphyromonas gingivalis is a periodontal pathogen whose long fimbriae (FimA) are classified into six genotypes (types I–V and Ib) based on the diversity of the fimA genes. FimA variations were previously shown to be related to the onset and development of adult periodontitis in a general population, while FimA were recently found to be critical mediators of initial biofilm formation. However, it is unclear if FimA variations have effects on biofilm features. Here, we compare the characteristic structures of homotypic biofilms developed by P. gingivalis strains with different FimA types.

Methods:  Biofilms were formed on saliva‐coated glass bottom wells in phosphate‐buffered saline and their structures were analysed using confocal laser scanning microscopy. Furthermore, the biovolumes of the biofilms were quantified with a three‐dimensional fluorophotometric method.

Results:  Biofilm structures formed by the six representative FimA‐type strains apparently differed. Type I and Ib P. gingivalis formed biofilms with a dense basal monolayer and dispersed microcolonies, whereas those formed by types II, III and IV strains had markedly luxuriant biofilms filled with widely clumped and tall colonies, and their biovolumes were significantly greater than those of types I and Ib. These characteristic features were confirmed to be closely related to FimA type in assays that utilized fimA‐substituted mutants from type I to II and those from type II to I.

Conclusion:  Our results suggest that FimA variations have effects on the structures of biofilms formed by P. gingivalis, which may be an important factor in the pathogenesis of periodontitis.
Examination of concordance between maternal and youth reports in the diagnosis of pediatric bipolar disorder
Authors: Biederman Joseph , Petty Carter R , Wilens Timothy E , Spencer Thomas , Henin Aude , Faraone Stephen V , Mick Eric , Monuteaux Michael C , Kenealy Deborah , Mirto Tara , Wozniak Janet
Published: April 2009

Objective:  While concordance between mother and child report continues to be the gold standard in the assessment of pediatric bipolar disorder, uncertainty develops when a mother’s report is not endorsed by the youth. To this end we compared discordant (mother positive and youth negative) and concordant (mother and youth positive) cases.

Methods:  Subjects were 98 adolescents (12–19 years of age) derived from family studies of bipolar disorder in youth who had both self‐reported and mother‐reported assessments. Comparisons were made between discordant (n = 35) and concordant (n = 59) cases on a wide range of clinical correlates.

Results:  Mothers in both groups reported similar rates of symptoms of mania and depression. Within the concordant group, mothers and youth reported similar rates of symptoms of mania. There were no differences between the concordant and discordant groups in onset, duration, or impairment of mania, rates of psychiatric hospitalization, cognitive variables, or rates of disorders in family members.

Conclusions:  The similarities between discordant and concordant reports in symptomatology of mania and depression, rates of comorbidities, treatment needs, and other clinical correlates suggest that a mother‐based diagnosis of mania should not be discounted in discrepant cases in which the youth fails to endorse the diagnosis.
Canadian Network for Mood and Anxiety Treatments (CANMAT) and International Society for Bipolar Disorders (ISBD) collaborative update of CANMAT guidelines for the management of patients with bipolar disorder: update 2009
Authors: Yatham Lakshmi N , Kennedy Sidney H , Schaffer Ayal , Parikh Sagar V , Beaulieu Serge , O'Donovan Claire , MacQueen Glenda , McIntyre Roger S , Sharma Verinder , Ravindran Arun , Young L Trevor , Young Allan H , Alda Martin , Milev Roumen , Vieta Eduard , Calabrese Joseph R , Berk Michael , Ha Kyooseob , Kapczinski Flávio
Published: April 2009

The Canadian Network for Mood and Anxiety Treatments (CANMAT) published guidelines for the management of bipolar disorder in 2005, with a 2007 update. This second update, in conjunction with the International Society for Bipolar Disorders (ISBD), reviews new evidence and is designed to be used in conjunction with the previous publications.

The recommendations for the management of acute mania remain mostly unchanged. Lithium, valproate, and several atypical antipsychotics continue to be first‐line treatments for acute mania. Tamoxifen is now suggested as a third‐line augmentation option. The combination of olanzapine and carbamazepine is not recommended. For the management of bipolar depression, lithium, lamotrigine, and quetiapine monotherapy, olanzapine plus selective serotonin reuptake inhibitor (SSRI), and lithium or divalproex plus SSRI/bupropion remain first‐line options. New data support the use of adjunctive modafinil as a second‐line option, but also indicate that aripiprazole should not be used as monotherapy for bipolar depression. Lithium, lamotrigine, valproate, and olanzapine continue to be first‐line options for maintenance treatment of bipolar disorder. New data support the use of quetiapine monotherapy and adjunctive therapy for the prevention of manic and depressive events, aripiprazole monotherapy for the prevention of manic events, and risperidone long‐acting injection monotherapy and adjunctive therapy, and adjunctive ziprasidone for the prevention of mood events.

Bipolar II disorder is frequently overlooked in treatment guidelines, but has an important clinical impact on patients’ lives. This update provides an expanded look at bipolar II disorder.
Selective deficits in semantic verbal fluency in patients with a first affective episode with psychotic symptoms and a positive history of mania
Authors: Kravariti Eugenia , Reichenberg Abraham , Morgan Kevin , Dazzan Paola , Morgan Craig , Zanelli Jolanta W , Lappin Julia M , Doody Gillian A , Harrison Glynn , Jones Peter B , Murray Robin M , Fearon Paul
Published: April 2009

Objectives:  Neurocognitive dysfunction is likely to represent a trait characteristic of bipolar disorder, but the extent to which it comprises ‘core’ deficits as opposed to those secondary to longstanding illness or intellectual decline is unclear. We investigated neuropsychological performance in an epidemiologically derived sample of patients with a first affective episode with psychotic symptoms and a positive history of mania, compared to community controls.

Methods:  Using a nested case‐control, population‐based study, measures of episodic and working memory, executive function, processing speed, and visual‐spatial perception were compared between 35 patients with a first affective episode with psychotic symptoms and a positive history of mania, and 274 community controls, as well as a subgroup of 105 controls matched on current IQ (‘good’ versus ‘poor’) and IQ trajectory (‘stable’, ‘declined’, or ‘improved’) with the patients (three controls per case).

Results:  Compared to the extended control sample, probands showed a suggestive deficit in short‐term verbal recall, and a significant deficit in semantic fluency. Only the latter was detectable in the comparison with the IQ‐matched controls. All other neurocognitive domains showed intact performance or nonsignificant deficits of small effect sizes compared to both control groups. Semantic fluency showed no association with symptoms or duration of untreated illness.

Conclusions:  Patients with a first affective episode with psychotic symptoms and a positive history of mania show an isolated, selective deficit in semantic verbal fluency, against a background of generally preserved neurocognitive function. This pattern seems to contrast with the more widespread neuropsychological dysfunction seen in schizophrenia.
Valproate activates the Notch3/c‐FLIP signaling cascade: a strategy to attenuate white matter hyperintensities in bipolar disorder in late life?
Authors: Yuan Peixiong , Salvadore Giacomo , Li Xiaoxia , Zhang Lei , Du Jing , Chen Guang , Manji Husseini K
Published: April 2009

Objectives:  Increased prevalence of deep white matter hyperintensities (DWMHs) has been consistently observed in patients with geriatric depression and bipolar disorder. DMWHs are associated with chronicity, disability, and poor quality of life. They are thought to be ischemic in their etiology and may be related to the underlying pathophysiology of mood disorders in the elderly. Notably, these lesions strikingly resemble radiological findings related to the cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephelopathy (CADASIL) syndrome. CADASIL arises from mutations in Notch3, resulting in impaired signaling via cellular Fas‐associated death domain‐like interleukin‐1‐beta‐converting enzyme‐inhibitory protein (c‐FLIP) through an extracellular signal‐regulated kinase (ERK)‐dependent pathway. These signaling abnormalities have been postulated to underlie the progressive degeneration of vascular smooth muscle cells (VSMC). This study investigates the possibility that the anticonvulsant valproate (VPA), which robustly activates the ERK mitogen‐activated protein kinase (MAPK) cascade, may exert cytoprotective effects on VSMC through the Notch3/c‐FLIP pathway.

Methods:  Human VSMC were treated with therapeutic concentrations of VPA subchronically. c‐FLIP was knocked down via small interfering ribonucleic acid transfection. Cell survival, apoptosis, and protein levels were measured.

Results:  VPA increased c‐FLIP levels dose‐ and time‐dependently and promoted VSMC survival in response to Fas ligand‐induced apoptosis in VSMC. The anti‐apoptotic effect of VPA was abolished by c‐FLIP knockdown. VPA also produced similar in vivo effects in rat brain.

Conclusions:  These results raise the intriguing possibility that VPA may be a novel therapeutic agent for the treatment of CADASIL and related disorders. They also suggest that VPA might decrease the liability of patients with late‐life mood disorders to develop DWMHs.
Clinical characteristics of unipolar disorder and bipolar disorder according to the lifetime presence of recurrent panic attacks
Authors: Forty Liz , Smith Daniel , Jones Lisa , Jones Ian , Caesar Sian , Cooper Carly , Fraser Christine , Gordon‐Smith Katherine , Hyde Sally , Farmer Anne , McGuffin Peter , Craddock Nick
Published: April 2009

Objectives:  The frequent comorbidity of panic and affective disorders has been described in previous studies. However, it is not clear how panic disorder comorbidity in unipolar disorder and bipolar disorder is related to illness course.

Methods:  We compared lifetime clinical characteristics of illness and items of symptomatology in samples of individuals with bipolar I disorder (n = 290) and unipolar disorder (n = 335) according to the lifetime presence of recurrent panic attacks.

Results:  We found significant differences in clinical course of illness characteristics that were shared across the unipolar and bipolar samples according to the lifetime presence of panic attacks. We also found a number of differences according to the presence of panic attacks that may be specific to the diagnostic group.

Conclusions:  Distinguishing patients who have mood disorder diagnoses, especially bipolar I disorder, according to the lifetime presence of panic attacks may not only be of use in clinical practice, but may also be informative for aetiological research, such as molecular genetic studies.
A comparison of ultrasound‐guided supraclavicular and infraclavicular blocks for upper extremity surgery
Authors: KOSCIELNIAK‐NIELSEN Z. J. , FREDERIKSEN B. S. , RASMUSSEN H. , HESSELBJERG L.
Published: April 2009

Background: Ultrasound (US)‐guided supraclavicular or infraclavicular blocks are commonly used for upper extremity surgery. The aims of this randomized study were to compare the block performance and onset times, effectiveness, incidence of adverse events and patient's acceptance of US‐guided supraclavicular or infraclavicular blocks. We hypothesized that the supraclavicular approach, being more superficial and easier to visualize using a 10 MHz transducer, will produce a faster and a more extensive sensory block.

Methods: One hundred and twenty patients were randomized to two equal groups: supraclavicular (S) and infraclavicular (I). Each patient received a mixture containing equal volumes of ropivacaine 7.5 mg/ml and mepivacaine 20 mg/ml with adrenaline 5 μg/ml, 0.5 ml/kg body weight (minimum 30 ml, maximum 50 ml). The sensory score (anaesthesia – 2 points, analgesia – 1 point and pain – 0 point) of the seven terminal nerves was assessed every 10 min. Patients were declared ready for surgery when they had an effective surgical block – anaesthesia or analgesia of the five nerves below the elbow. Thirty minutes after the block, the unblocked nerves were supplemented. The block performance and latency times, surgical effectiveness, adverse events and patient's acceptance were recorded.

Results: Significantly more patients in the I group were ready for surgery 20 and 30 min after the block. The mean block performance time was 5.7 min in the S group and 5.0 min in the I group (NS). Block effectiveness was superior in the I group: 93% vs. 78% in the S group (P=0.017). The S group patients had a significantly poorer block of the median and ulnar nerves, but a better block of the axillary nerve. Sensory scores at 10, 20 and 30 min were not significantly different. Thirty‐two patients in the S group vs. nine patients in the I group experienced transient adverse events (P<0.0001). Patients' acceptance of the block was similar in both groups.

Conclusions: Infraclavicular block had a faster onset, better surgical effectiveness and fewer adverse events. Block performance time and patients' acceptance of the procedure were similar in both groups.
Prescription pattern of codeine for non‐malignant pain: a pharmacoepidemiological study from the Norwegian Prescription Database
Authors: FREDHEIM O. M. S. , SKURTVEIT S. , MOROZ A. , BREIVIK H. , BORCHGREVINK P. C.
Published: April 2009

Background: Opioid prescription for pain relief is increasing. Codeine is the dominating opioid in several European countries, with Norway being among the highest codeine users.

Aim: To determine whether codeine is primarily used for acute pain or whether there is a prescription pattern indicating problematic opioid use.

Methods: All pharmacies in Norway are obliged to submit data electronically to the Norwegian Prescription Database at the Norwegian Institute of Public Health on all dispensed prescriptions. Because all prescriptions are identified with a unique person identifier, it is possible to identify all prescriptions to one subject. All subjects who had prescription(s) of codeine dispensed to them in 2004, 2005 or 2006 are included in the study.

Results: 385 190 Norwegian persons had at least one prescription of codeine dispensed to them due to non‐cancer pain in 2005, corresponding to a 1‐year periodic prevalence of 8.3%. 223 778 (58%) received only one prescription in 2005, 121 025 (31%) received more than one prescription but <120 defined daily doses (DDDs), 30 939 (8%) received between 120 and 365 DDDs, 7661 (2%) between 365 and 730 DDDs, while only 1787 (0.5%) exceeded the maximum recommended dose of 730 DDDs. In the latter group, co‐medication with benzodiazepines (65%) and carisoprodol (45%) was prevalent.

Conclusion: About one in 10 adult persons in Norway were dispensed codeine in 2005. A majority (58%) received codeine only once, most likely for acute pain, whereas a small minority (0.5%) had a prescription pattern indicating problematic opioid use.