bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of monoclonal antibody (P504S) and polyclonal antibody to alpha methylacyl‐CoA racemase (AMACR) in the work‐up of prostate cancer
Authors: Kunju L P , Chinnaiyan A M , Shah R B 
Published: October 2005

Aim : Studies using a monoclonal (P504S) and a polyclonal antibody (p‐AMACR) to α‐methylacyl‐CoA racemase (AMACR) have shown variable expression in prostate cancer (PCa). The goal is to compare the sensitivity of both antibodies in PCa and evaluate their utility in the work‐up of atypical prostate needle biopsies (NBXs).Methods and results : A tissue microarray (TMA) with 248 samples of benign prostate, high‐grade prostatic intraepithelial neoplasia (HGPIN) and PCa samples, 20 NBXs with minute PCa and 32 NBXs with ‘atypical’ foci were stained with P504S and p‐AMACR. Ninety percent of PCa (76/76 TMA, 16/20 NBXs) showed predominantly strong p‐AMACR expression while 87% (65/69 TMA, 16/20 NBXs) showed variable P504S expression (sensitivity 90% versus 87%, P = 0.10). In HGPIN, P504S and p‐AMACR were positive in 77% and 91% of samples, respectively. In the ‘atypical’ NBXs group, 53% were classified as PCa, 12% benign and 35% atypical, suspicious for PCa, after review of the basal marker. Of atypical, suspicious for PCa, P504S/p‐AMACR helped convert the diagnosis to PCa in 5/11 (45%) cases, where, despite negative basal cell markers, morphology was less than optimal.Conclusions : Differences between P504S and p‐AMACR appear marginal and clinically insignificant. AMACR is negative in a subset of unequivocal minute PCa with both antibodies. However, when utilized in proper context, AMACR may offer significant advantage in converting an ‘atypical’ diagnosis to PCa where morphology and basal markers are less than optimal in resolving the diagnosis.
Gene clusters involved in anaerobic benzoate degradation of Geobacter metallireducens
Authors: Wischgoll Simon , Heintz Dimitri , Peters Franziska , Erxleben Anika , Sarnighausen Eric , Reski Ralf , Van Dorsselaer Alain , Boll Matthias 
Published: October 2005

SummaryThe degradation of aromatic compounds follows different biochemical principles in aerobic and anaerobic microorganisms. While aerobes dearomatize and cleave the aromatic ring by oxygenases, facultative anaerobes utilize an ATP‐dependent ring reductase for the dearomatization of the activated key intermediate benzoyl‐coenzyme A (CoA). In this work, the aromatic metabolism was studied in the obligately anaerobic model organism Geobacter metallireducens. The gene coding for a putative carboxylic acid‐CoA ligase was heterologously overexpressed and the gene product was characterized as a highly specific benzoate‐CoA ligase catalysing the initial step of benzoate metabolism. However, no evidence for the presence of an ATP‐dependent benzoyl‐CoA reductase as observed in facultative anaerobes was obtained. In a proteomic approach benzoate‐induced proteins were identified; the corresponding genes are organized in two clusters comprising 44 genes. Induction of representative genes during growth on benzoate was confirmed by reverse transcription polymerase chain reaction. The results obtained suggest that benzoate is activated to benzoyl‐CoA, which is then reductively dearomatized to cyclohexa‐1,5‐diene‐1‐carbonyl‐CoA, followed by β‐oxidation reactions to acetyl‐CoA units, as in facultatively anaerobic bacteria. However, in G. metallireducens the process of reductive benzene ring dearomatization appears to be catalysed by a set of completely different protein components comprising putative molybdenum and selenocysteine containing enzymes.
Iron‐reducing bacteria unravel novel strategies for the anaerobic catabolism of aromatic compounds
Authors: Carmona Manuel , Díaz Eduardo 
Published: October 2005

SummaryAlthough the aerobic degradation of aromatic compounds has been extensively studied in many microorganisms, the anaerobic mineralization of the aromatic ring is a more recently discovered microbial capacity on which very little information is available from facultative anaerobic bacteria. In this issue of Molecular Microbiology, Wischgoll and colleagues use proteomic and reverse‐transcription polymerase chain reaction (PCR) approaches to identify for the first time the gene clusters involved in the central pathway for the catabolism of aromatic compounds in Geobacter metallireducens, a strictly anaerobic iron‐reducing bacterium. This work highlights that the major difference in anaerobic benzoate metabolism of facultative and strictly anaerobic bacteria is the reductive process for dearomatization of benzoyl‐CoA. The authors propose that a new type of benzoyl‐CoA reductase, comprising molybdenum‐ and selenocysteine‐containing proteins, is present in strictly anaerobic bacteria. This work paves the way to fundamental studies on the biochemistry and regulation of this new reductive process and provides the first genetic clues on the anaerobic catabolism of benzoate by strict anaerobes.
Release of the type I secreted α‐haemolysin via outer membrane vesicles from Escherichia coli
Authors: Balsalobre Carlos , Silván Jose Manuel , Berglund Stina , Mizunoe Yoshimitsu , Uhlin Bernt Eric , Wai Sun Nyunt 
Published: October 2005

SummaryThe α‐haemolysin is an important virulence factor commonly expressed by extraintestinal pathogenic Escherichia coli. The secretion of the α‐haemolysin is mediated by the type I secretion system and the toxin reaches the extracellular space without the formation of periplasmic intermediates presumably in a soluble form. Surprisingly, we found that a fraction of this type I secreted protein is located within outer membrane vesicles (OMVs) that are released by the bacteria. The α‐haemolysin appeared very tightly associated with the OMVs as judged by dissociation assays and proteinase susceptibility tests. The α‐haemolysin in OMVs was cytotoxically active and caused lysis of red blood cells. The OMVs containing the α‐haemolysin were distinct from the OMVs not containing α‐haemolysin, showing a lower density. Furthermore, they differed in protein composition and one component of the type I secretion system, the TolC protein, was found in the lower density vesicles. Studies of natural isolates of E. coli demonstrated that the localization of α‐haemolysin in OMVs is a common feature among haemolytic strains. We propose an alternative pathway for the transport of the type I secreted α‐haemolysin from the bacteria to the host cells during bacterial infections.
Mucin–Pseudomonas aeruginosa interactions promote biofilm formation and antibiotic resistance
Authors: Landry Rebecca M. , An Dingding , Hupp Joseph T. , Singh Pradeep K. , Parsek Matthew R. 
Published: October 2005

Summary Pseudomonas aeruginosa is an opportunistic pathogen that causes chronic lung infections in people suffering from cystic fibrosis (CF). In CF airways, P. aeruginosa forms surface‐associated communities called biofilms. Compared with free‐swimming cultures, biofilms resist clearance by the host immune system and display increased resistance to antimicrobial agents. In this study we developed a technique to coat surfaces with molecules that are abundant in CF airways in order to investigate their impact on P. aeruginosa biofilm development. We found that P. aeruginosa biofilm development proceeds differently on surfaces coated with the glycoprotein mucin compared with biofilm development on glass and surfaces coated with actin or DNA. Biofilms formed on mucin‐coated surfaces developed large cellular aggregates and had increased tolerance to the antibiotic tobramycin compared with biofilms grown on glass. Analysis of selected mutant backgrounds in conjunction with time‐lapse microscopy revealed that surface‐associated motility was blocked on the mucin surface. Furthermore, our data suggest that a specific adhesin–mucin interaction immobilizes the bacterium on the surface. Together, these experiments suggest that mucin, which may serve as an attachment surface in CF airways, impacts P. aeruginosa biofilm development and function.
Characterization of the putative operon containing arylamine N‐acetyltransferase (nat) in Mycobacterium bovis BCG
Authors: Anderton Matthew C. , Bhakta Sanjib , Besra Gurdyal S. , Jeavons Peter , Eltis Lindsay D. , Sim Edith 
Published: October 2005

Summary Mycobacterium bovis BCG and Mycobacterium tuberculosis possess a single arylamine N‐acetyltransferase whose gene is predicted to occur within a six‐gene operon. Deletion of the nat gene caused an extended lag phase in M. bovis BCG and a cell morphology associated with an altered pattern of cell wall mycolates. Analysis of cDNA from M. bovis BCG shows that during in vitro growth all the genes in the putative nat operon are expressed and the open reading frames are contiguous, supporting the existence of an operon. Two genes in the operon, Mb3599c and Mb3600c, are predicted to encode homologues of enzymes annotated as a 2,3‐dihydroxybiphenyl 1,2‐dioxygenase (bphC5) and a 2‐hydroxy‐6‐oxo‐6‐phenylhexa‐2,4‐dienoate hydrolase (bphD2), respectively, in Rhodococcus RHA1. As predicted, M. bovis BCG cell lysates metabolized the BphC substrate 2,3‐dihydroxybiphenyl (2,3‐DHB) to 2‐hydroxy‐6‐oxo‐6‐phenylhexa‐2,4‐dienoic acid (HOPDA), a BphD substrate, which was subsequently hydrolysed. Immunoprecipitation of the BphD homologue from these lysates led to an accumulation of HOPDA. M. bovis BCG growth on both solid and liquid media was inhibited with either 2,3‐DHB or an inhibitor of BphC, 3‐chlorocatechol (3‐CC). In addition, incubation with 2,3‐DHB affects the lipid composition of the cell wall resulting in a diminished level of mycolates and an altered cell morphology similar to the Δnat strain. We propose the enzymes encoded by the putative operon have a similar endogenous role to that of the NAT enzyme and are part of a pathway important for cell wall synthesis.
Specific immunotherapy with SQ standardized grass allergen tablets in asthmatics with rhinoconjunctivitis
Authors: Dahl R. , Stender A. , Rak S. 
Published: October 2005

Background:  The best way to prevent allergy symptoms is to treat the allergic condition. Specific immunotherapy with grass allergen tablets 75 000 SQ‐T (Grazax®Phleum pratense, ALK‐Abelló) is safe and efficacious in rhinoconjunctivitis patients. As rhinoconjunctivitis often co‐exists with asthma, we aimed to confirm safety and efficacy in grass allergic subjects with asthma and rhinoconjunctivitis.Methods:  A randomized, double‐blind, placebo‐controlled, multicentre trial was performed 10–14 weeks prior to and during the grass pollen season 2004. About 114 subjects were randomized 2 : 1 to grass allergen tablets or placebo. The primary end points were average asthma medication and symptom scores during the grass pollen season, and secondary variables were average rhinoconjunctivitis symptom and medication scores during the grass pollen season. Additionally, number of well days was defined post hoc.Results:  Differences in asthma medication and symptom scores between the treatment groups were negligible. The mean difference in asthma medication score was below 0.1 and 0.3 for asthma symptom score [a single inhalation of salbutamol (200 μg) was scored 2]. No serious adverse events were reported.A reduction in rhinoconjunctivitis symptom score of 37% (P = 0.004) and a 41% (P = 0.036) reduction in medication score was found in the grass pollen season for subjects treated with the grass allergen tablet compared with placebo. Well days increased by 54% (P = 0.002).Conclusions:  Self‐administration of the grass allergen tablet was safe. The treatment did not impair asthma control and confirmed considerable symptom prevention and reduced medication use. It addresses the allergic condition and represents a baseline treatment for grass pollen allergy.
CD4+CD30+ T cells perpetuate IL‐5 production in Dermatophagoides pteronyssinus allergic patients
Authors: Garfias Y. , Ortiz B. , Hernández J. , Magaña D. , Becerril‐Angeles M. , Zenteno E. , Lascurain R. 
Published: October 2005

Background:  Airway allergic diseases are regulated by interleukin (IL)‐5, which causes infiltration of eosinophils into the bronchial epithelium, and by IL‐4 which increases serum immunoglobulin E (IgE) production and promotes CD30 expression on Th cells. CD30 generates a costimulatory signal involved in apoptosis or cell proliferation, depending on the microenvironment. Our aims were: (i) to analyze if CD4+CD30+ T cells from allergic patients proliferate in response to Dermatophagoides pteronyssinus, and (ii) if upon stimulation this cell population produces IL‐4 and IL‐5.Methods:  Peripheral blood mononuclear cell (PBMC) from 17 allergic rhinitis and mild allergic asthma patients and 12 healthy nonallergic individuals were stimulated with allergen in the presence or absence of anti‐IL‐4, anti‐IL‐5 or anti‐IL‐4Rα monoclonal antibodies (mAbs). TdT‐mediated dUTP nick end‐labeling (TUNEL) assay, 7‐aminoactinomycin‐D (7‐AAD) intercalation, and flow cytometry were used to determine the CD4+CD30+ blasts percentage, cell proliferation, apoptosis, and intracellular cytokines after 7 culture days.Results:  Cell proliferation induced with allergen showed that 90% of the allergen‐stimulated blasts were CD4+, 50% of which were CD30+. Allergen‐stimulated PBMC showed a progressive increase (mean: from 7% to 23%) of CD4+CD30+IFN‐γ+ and CD4+CD30+IL‐4+ blasts which diminished (mean: 6%) after 5 culture days. In contrast, CD4+CD30+IL‐5+ blasts showed a continuous progression (from 12% to 24%) that maintained after 7 culture days. The vast majority of CD4+CD30+ blasts were negative to 7‐AAD or TUNEL. Additionally, a significant decrease (34%) was observed in the number of CD4+CD30+ blasts when IL‐4 was neutralized.Conclusions:  These data suggest that specific allergen stimulation of PBMC isolated from allergic patients generates a nonapoptotic CD4+CD30+ blast subset that produces IL‐5.
Malassezia sympodialis differently affects the expression of LL‐37 in dendritic cells from atopic eczema patients and healthy individuals
Authors: Agerberth B. , Buentke E. , Bergman P. , Eshaghi H. , Gabrielsson S. , Gudmundsson G. H. , Scheynius A. 
Published: October 2005

Background:  Atopic eczema (AE) is a multifactorial disease, which has increased in prevalence. The skin‐colonizing yeast Malasezzia sympodialis can induce IgE‐ and T‐cell reactivity in patients with AE. LL‐37 is an endogenous peptide antibiotic belonging to the cathelicidin family. The aim of this study was to examine whether exposure to M. sympodialis would affect the expression of LL‐37 in dendritic cells.Methods:  The presence of LL‐37 was analyzed in monocyte‐derived dendritic cells (MDDCs) generated from healthy individuals and patients with AE by Western blotting and the corresponding cDNA by real‐time quantitative RT‐PCR. Antibacterial activity was measured with an inhibition zone assay in fractions after reverse phase chromatography.Results:  For the first time we here present data, showing that LL‐37 is produced by MDDCs. Notably, the secretion of LL‐37 was substantially enhanced in M. sympodialis‐exposed MDDCs generated from patients with a high degree of eczema, as measured by SCORAD, compared to healthy controls and patients with a low SCORAD. The relative expression of LL‐37 transcript in MDDCs generated from patients was up‐regulated after 1 h of exposure to M. sympodialis and declined gradually at the time points analyzed, whereas the transcription was unaffected in the MDDCs of healthy controls.Conclusions:  Our results suggest that M. sympodialis can trigger the innate immune response differently in patients with AE and healthy individuals. The enhanced LL‐37 secretion from the MDDCs in the patients with AE may reflect the severity of their inflammatory response to M. sympodialis.
Motivation, recruitment and retention of voluntary non‐remunerated blood donors: a survey‐based questionnaire study
Authors: Misje A. H. , Bosnes V. , Gåsdal O. , Heier H. E. 
Published: October 2005

Background and Objectives  The aim of this study was to establish which motivational and socio‐demographic factors are important for the development of a long‐term commitment as a voluntary, non‐remunerated blood donor.Study Design and Methods  A cross‐sectional sample survey of active blood donors in Oslo, Norway, was conducted. Donors filled in a self‐administered questionnaire during donation. Data on motivation were analysed using factor analysis.Results  The blood donors’ socio‐demographic characteristics were found to be similar to those of the population as a whole. The single, most important, recruitment channel was the influence of active blood donors. Five dimensions of blood‐donor motivation were identified with factor analysis. These were: altruism and empathy; social reasons (such as the influence of friends and family); strengthening of one's self‐esteem; positive experiences associated with donation; and a moral obligation to donate. Support for statements on altruistic motives for donation was strong and similar in long‐time and short‐time donors. In contrast, short‐time donors were more likely to be motivated by factors related to self‐esteem than were long‐term donors.Conclusion  The ‘good habit’ of continued blood donation seems not to be exclusively linked to a high degree of reported other‐regarding (‘altruistic’) reasons, but also to a combination of motives, including some modestly self‐regarding motives.
Optimal sampling time after preparation of platelet concentrates for detection of bacterial contamination by quantitative real‐time polymerase chain reaction
Authors: Mohammadi T. , Pietersz R. N. I. , Scholtalbers L. A. H. , Vandenbroucke‐Grauls C. M. J. E. , Savelkoul P. H. M. , Reesink H. W. 
Published: October 2005

Background and Objectives  A universal quantitative real‐time polymerase chain reaction (PCR), based on bacterial 16S rDNA, to detect bacterial contamination of platelet concentrates (PCs), was developed previously and compared with automated culturing. In the present study, this real‐time PCR method was evaluated to determine the optimal sampling time for screening of bacterial contamination in PCs.Materials and Methods  Routinely prepared PCs were spiked with suspensions of Escherichia coli, Bacillus cereus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Propionibacterium acnes to 1, 10 and 100 colony‐forming units (CFU)/ml and stored at room temperature for 7 days. The presence of bacteria in these PCs was monitored by quantitative real‐time PCR. As a reference method (additional control), BacT/Alert automated culturing was used. For PCR, 1‐ml aliquots were drawn from all (spiked) PCs on days 0, 1, 2, 3, 6 and 7 of storage. As a control, triplicate samples (10 ml) were inoculated into aerobic and anaerobic BacT/Alert culture bottles immediately after spiking (day 0) and after storage for 1, 2, 3, 6 or 7 days.Results  With quantitative real‐time PCR, all bacterial species tested were reproducibly detected on day 1 after spiking at original concentrations of 10 and 100 CFU/ml. Bacteria were also detected on day 1 from PCs spiked with an initial concentration of 1 CFU/ml, except for E. coli, which was detected in only one of the three samples and P. aeruginosa, for which analysis was not performed on day 1. With the reference method, bacteria were detected in culture bottles (inoculated on day 0) within a mean time of 20·1 h, with the exception of P. acnes which was detected at a mean time of 102·3 and 49·3 h (for original spiking concentrations of 10 and 100 CFU/ml respectively).Conclusions  PCR enables the rapid detection of low initial numbers of bacteria in PCs. For reliable detection, our results support that sampling of PCs for real‐time PCR screening should not be carried out earlier than 1 day after preparation (48 h after blood collection). Importantly, the real‐time PCR approach has the potential to be used before the release of PCs from the blood centre or shortly before they are transfused in the hospital.
Extending the Box–Cox Transformation to the Linear Mixed Model
Authors: Gurka Matthew J. , Edwards Lloyd J. , Muller Keith E. , Kupper Lawrence L. 
Published: October 2005

SummaryFor a univariate linear model, the Box–Cox method helps to choose a response transformation to ensure the validity of a Gaussian distribution and related assumptions. The desire to extend the method to a linear mixed model raises many vexing questions. Most importantly, how do the distributions of the two sources of randomness (pure error and random effects) interact in determining the validity of assumptions? For an otherwise valid model, we prove that the success of a transformation may be judged solely in terms of how closely the total error follows a Gaussian distribution. Hence the approach avoids the complexity of separately evaluating pure errors and random effects. The extension of the transformation to the mixed model requires an exploration of its potential effect on estimation and inference of the model parameters. Analysis of longitudinal pulmonary function data and Monte Carlo simulations illustrate the methodology discussed.
The effect of low‐dose ritonavir monotherapy on fasting serum lipid concentrations
Authors: Shafran SD , Mashinter LD , Roberts SE 
Published: October 2005

ObjectivesRitonavir (RTV) at doses of 400 mg twice a day (bid) or higher adversely affects serum lipids. However, the effect of RTV 100 mg bid on serum lipids is unknown. We conducted a study to evaluate the effect of RTV 100 mg bid on fasting serum lipid profiles in HIV‐negative healthy volunteers.MethodsRitonavir 100 mg bid was administered for 14 days to 20 healthy HIV‐seronegative adults with normal serum lipids. After a 7‐day washout, lopinavir/ritonavir (LPV/RTV) 400/100 mg bid was administered for 14 days. Fasting serum lipid parameters were measured twice at baseline, after 14 days of RTV, and after 14 days of LPV/RTV, and comparisons were made at each time‐point for levels of total cholesterol, low‐density lipoprotein (LDL) cholesterol, high‐density lipoprotein (HDL) cholesterol, the total/HDL cholesterol ratio and triglycerides.ResultsAfter 14 days of RTV 100 mg bid, total cholesterol level increased by 10.2% (P<0.001), LDL cholesterol level increased by 16.2% (P<0.001), triglyceride levels increased by 26.5% (P<0.001), HDL cholesterol level decreased by 5.4% (P<0.01) and the total/HDL cholesterol ratio increased by 17.3% (P<0.001). The addition of LPV 400 mg bid to RTV 100 mg bid resulted in no significant further changes in LDL cholesterol or triglyceride level or total/HDL cholesterol ratio, but there were significant increases in both total cholesterol (8.0% increase; P=0.007) and HDL cholesterol levels (6.7% increase; P=0.008).ConclusionsRitonavir dosed at 100 mg bid significantly increased the concentration of total cholesterol, LDL cholesterol, total/HDL cholesterol ratio and triglycerides and reduced HDL cholesterol concentration. The addition of LPV 400 mg bid to RTV 100 mg bid further increased both total and HDL cholesterol levels without affecting the total/HDL ratio.