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The purpose of this study was to compare compliance with recommendations and clinical outcomes between formal and informal infectious disease specialist consultations. Six hundred twenty-seven consecutive adult inpatients who received an infectious disease consultation in a university-affiliated hospital were included. After adjusting for quintile of propensity score, we compared compliance with the consultant's recommendations and clinical outcomes for 443 (70.7%) and 184 (29.3%) formal and informal consultations. Informal and formal consultations were associated with comparable levels of compliance with recommendations for antimicrobial treatment (86.5% vs 88.9%; adjusted odds ratio [aOR], 0.63; 95% confidence interval, 0.34-1.14; P = 0.13) and diagnostic or monitoring tests (72.6% vs 72.0%; aOR, 0.91 [0.53-1.57]; P = 0.73). The rates of early clinical improvement (58.2% vs 58.6%; aOR, 1.11 [0.70-1.74]; P = 0.66), subsequent consultation (34.2% vs 36.3%; aOR, 0.80 [0.53-1.21]; P = 0.29), in-hospital mortality (4.9% vs 8.4%; aOR, 0.55 [0.24-1.24]; P = 0.15), and the median length of stay (23 vs 20 days; aOR of discharge, 0.90 [0.74-1.10]; P = 0.30) did not differ depending on the type of consultation. This study provides observational evidence that informal consultations result in levels of compliance with recommendations comparable to formal consultations, without compromising patient safety. Further study is needed to refine the criteria for requesting or providing informal rather than formal consultations. [ABSTRACT FROM AUTHOR]

The objective of this study is to determine whether the likelihood of returning for routine breast cancer screening differed for false-positive cases depending on the diagnostic work-up. Using the original data from a French population-based breast cancer screening program, we compared the attendance rates at the subsequent round of screening for 16,946 and 1,127 participants who received negative (i.e., American College of Radiology, ACR, categories 1-2) and false-positive mammograms, respectively. False-positive mammograms were categorized ACR 0 (i.e., warranting additional imaging evaluation), 3 (i.e., warranting clinical and imaging follow-up), and 4-5 (i.e., warranting biopsy). We estimated the odds ratios of attendance at subsequent screening round using logistic regression, adjusting for age and history of previous mammography. The attendance rates at the subsequent screening round were 80.6% for women who received negative mammograms versus 69.6, 74.3, and 70.1% for women who received false-positive mammograms warranting additional imaging evaluation, clinical and imaging follow-up, or biopsy, respectively. In comparison to women who received negative mammograms, the corresponding adjusted odds ratios of returning for routine screening were 0.6 [95% confidence interval (CI) 0.4-0.8], 0.8 (95% CI 0.6-0.9), and 0.6 (95% CI 0.4-0.8). No significant differences were found in odds ratios of attendance across ACR categories among women who received false-positive mammograms. Similar figures were observed for attending at least one of the two subsequent screening rounds. In conclusion, in comparison to women with normal or benign findings on index mammograms, false-positive cases warranting additional imaging evaluation, clinical and imaging follow-up, or biopsy had uniformly decreased odds of attending subsequent routine screening rounds. [ABSTRACT FROM AUTHOR]

We aimed to assess the appropriateness of routine blood culture testing parameters and antimicrobial therapy for patients with suspected bloodstream infection. We conducted a cross-sectional study of blood cultures registered in the microbiological laboratory at a university-affiliated hospital from 4 to 15 June 2007. Using a structured implicit chart review, two infectious disease specialists assessed the appropriateness of the testing parameters and antimicrobial therapy. Overall, 2,696 blood culture bottles were collected from 260 patients during their stay, including 955 bottles that were evaluated during the study period. The indication of fungal and bacterial blood cultures was rated as appropriate for 75% (95% confidence interval [CI], 65-83) and 91% (95% CI, 87-95) of patients. Only 45% (95% CI, 39-52) of patients had an adequate number of blood cultures (i.e., two to four). An optimal volume of blood (i.e., ≥10 mL) was inoculated in 13% (95% CI, 11-15) of adult bottles. Empirical antimicrobial therapy was appropriate for 60% (95% CI, 43-76) of patients with positive blood cultures. There is room for improvement regarding routine blood culture testing parameters and antimicrobial therapy. The effectiveness of multifaceted interventions in altering the appropriateness of blood culture parameters deserves further research. [ABSTRACT FROM AUTHOR]

BACKGROUND: Although older patients with restricted mobility are at increased risk for venous thromboembolism, they are under-represented in clinical trials evaluating prophylactic treatments against deep vein thrombosis (DVT). OBJECTIVE: To determine whether prolonged prophylaxis with low-molecular-weight heparin (LMWH) is associated with a lower rate of DVT in older patients with restricted mobility. METHODS: Two cross-sectional studies were conducted in 50 hospital-based, post-acute care facilities in France in 2001 and 2003. The studies included 1603 evaluable patients aged >or=65 years, including 866 LMWH users (median treatment duration 23 days; interquartile range 13-42) and 737 LMWH non-users. All patients underwent complete compression ultrasonography performed by board-certified vascular medicine physicians. The primary study outcome was proximal DVT. Propensity analyses were used to control for bias in LMWH treatment assignment. RESULTS: The rate of proximal DVT was 4% (35/866) and 5.7% (42/737) for LMWH users and non-users, respectively (p = 0.16). Prophylaxis with LMWH was associated with decreased odds of proximal DVT after adjusting for baseline characteristics (odds ratio [OR] 0.56; 95% CI 0.33, 0.95; p = 0.03) or quintile of propensity score (OR 0.58; 95% CI 0.35, 0.99; p = 0.04). In propensity matched analysis, 342 LMWH users were at decreased odds of proximal DVT compared with 342 non-users (OR 0.50; 95% CI 0.24, 1.00; p = 0.04). The decrease in proximal DVT was paralleled by a similar decrease in distal DVT. Compared with non-users, only high-risk dose users had decreased odds of DVT. CONCLUSIONs: In this observational study, prophylaxis with a high-risk dose of LMWH was associated with decreased odds of proximal DVT in older patients with restricted mobility. Further study is needed before recommending routine prophylaxis with LMWH in these patients. [ABSTRACT FROM AUTHOR]

An ORF of 1716 nucleotides, putatively encoding a DNA polymerase, was characterized in the mitochondrial genome of the edible basidiomycete Agrocybe aegerita. The complete gene, named Aa-polB, and its flanking regions were cloned and sequenced from three overlapping restriction fragments. Aa-polB is located between the SSU rDNA (5′ region) and a gene for tRNAAsn (3′ region), and is separated from these genes by two A+T-rich intergenic regions of 1048 (5′ region) and 3864 (3′ region) nucleotides, which lack repeated sequences of mitochondrial or plasmid origin. The deduced Aa-POLB protein shows extensive sequence similarity with the family B DNA polymerases encoded by genomes that rely on protein-primed replication (invertrons). The domains involved in the 3′→5′ exonuclease (Exo I to III) and polymerase (Pol I to Pol V) activities were localized on the basis of conserved sequence motifs. The alignment of the Aa-POLB protein (571 amino acids) with sequences of family B DNA polymerases from invertrons revealed that in Aa-POLB the N-terminal region preceding Exo I is short, suggesting a close relationship with the DNA polymerases of bacteriophages that have linear DNA. The Aa-polB gene was shown to be present in all wild strains examined, which were collected from a wide range of locations in Europe. As shown by RT-PCR, the Aa-polB gene is transcribed in the mitochondria, at a low but significant level. The likelihood of the coexistence of Aa-POLB and Pol γ in the A. aegerita mitochondrion is discussed in the light of recent reports showing the conservation of the nucleus-encoded Pol γ from yeast to human. [ABSTRACT FROM AUTHOR]

The Aa-Pri2 gene, specifically expressed during basidiocarp differentiation of the mushroom Agrocybe aegerita, was cloned. Sequence analysis showed a 525-nt ORF interrupted by three introns (51, 48, 54 bp). One TATA box (–94), two CAAT boxes (–382, –398) and one CT-rich motif (–149) were present. The transcription start point was located by primer extension at position –60. The hydrophobic Aa-PRI2 protein (123 aa) possessed a putative signal peptide (aa1–aa18), eight cysteine residues characteristic of the hydrophobin proteins, and one putative leucine-zipper domain (aa50–aa71). Hybridization of the genomic DNA of several edible mushrooms with the Aa-Pri2 cDNA, showed a 2.7-kb DNA fragment highly similar to the Aa-Pri2 gene in the species Agrocybe chaxingu; sequences showing slight hybridization signals were detected in Pleurotus eryngii, Lentinula edodes and Agaricus bisporus. [ABSTRACT FROM AUTHOR]

A procedure suitable for the extraction and mapping of total proteins from the basidiomycete, Agrocybe aegerita, was developed. A. aegerita mycelia were fragmented either with a Dangoumeau grinder, an X-press bomb or a sonicator and the efficiency of these three disruption methods were compared. The extraction buffer composition was optimized to avoid proteolytic activities. 2D-SDS-PAGE analysis of protein extracts showed that the rate of reproducibility depending on extractions and electrophoretic separations was always greater than 96% for all strains. The differences in efficiency observed between the breaking procedures indicate that the A. aegerita cell wall is more mechanically resistant than that of other basidiomycetes. The efficient action of protease inhibitors (PMSF and SDS) showed that A. aegerita mycelia contains numerous and/or highly active proteases. Reproducibility of protein extraction and separation methods allowed the establishment and the comparison of standard maps. Qualitative and quantitative variations in gene products between a wild dikaryotic strain and 11 homokaryotic strains from its progeny were examined. The genetic diversity, determined by comparing the distribution of proteic variations in 11 homokaryons from the same progeny, was comparable to that observed between co-isogenic homokaryons of another basidiomycete. [ABSTRACT FROM AUTHOR]

A gene ( Aa-Pri1) specifically expressed during fruiting initiation of the basidiomycete Agrocybe aegerita was cloned. The total length of the Aa-Pri1 gene was 492 bp including a class-II intron of 54 bp size located at nt +125; the open reading frame encoded for a 145-aa protein of 16 093 Da. CCAAT (–156) and TATAAAT (– 83) boxes, and T(A)5T(A)2 (+593) and T(A)3T(A)4T(A)6T (+608) putative polyadenylation sequences were identified. The putative transcription start point was located at position 49. The Aa-Pri1 transcript was abundant only during fruiting initiation and was undetectable in the other stages of development. The Aa-Pri1 protein was hydrophilic, with a 20-aa hydrophobic motif in the NH2-terminal part, determining a putative α-helix. Two putative glycosylation sites were identified. Aa-Pri1 protein activity may be controlled by the phosphorylation of several residues by different protein kinases. [ABSTRACT FROM AUTHOR]

A protein that binds Concanavalin A (Con A) was detected on Western blots of Spiroplasma citri proteins. Its apparent molecular weight was 84000. It was localized in the plasma membrane. Affinity chromatography on Con A-agarose was used to isolate this protein. The glycosylation inhibitor, tunicamycin, inhibits S. citri growth and seems to block the glycosylation of the Con A-binding protein. [ABSTRACT FROM AUTHOR]

DNA sequences of the basidiomycete Agrocybe aegerita were cloned in E. coli based on their ability to drive the expression of the bacterial promoterless tetracycline (Tc)-resistance gene. A 0.48% frequency of the cloned sequences promoted antibiotic-resistance. The sequence conferring the highest Tc resistance (40 μg/ml) was selected to drive the expression in E. coli of two other promoterless genes encoding chloramphenicol and neomycin resistance. One of the derivative vectors, pN13-A2, carrying a chimeric neomycin-resistance gene, was used to transform an A. aegerita neomycin-sensitive strain by protoplast electroporation. Transformation frequencies ranged from 1 to 2.8 transformants per μg of DNA per 10 viable cells, in a relatively high background of spontaneous-resistant colonies (2% of the surviving protoplasts). Molecular analyses showed that transformation had occurred by the integration of pN13-A2 sequences, either ectopically or at the resident locus carrying the A. aegerita promoter-like sequence, with probable molecular rearrangements. The nucleotide sequence of the promoter-like fragment revealed the presence of a CT motif that is known to be involved in a promoter function in some highly expressed genes of filamentous fungi. [ABSTRACT FROM AUTHOR]

In the ribosomal DNA unit of Pleurotus cornucopiae, the rDNA coding regions are in the order 5′, 5S-18S-5.8S-25S, 3′, with the 5′ location of the 5S gene differing from its 3′ location found in other basidiomycetes. The most discriminating probe used to study the rDNA polymorphism consisted of a fragment that included the 5S, 18S and part of the 5.8S and 25S genes flanking three intergenic sequences. A high degree of rDNA polymorphism was observed in the seven P. cornucopiae dikaryons studied. For the first time within a basidiomycete species, the restrictions maps distinguished two types of rDNA units (I and II). In each rDNA type, length variations in the external intergenic sequence IGS 1 located between the 25S and 5S genes allowed characterization of two different rDNA units in type I and four rDNA units in type II. This suggested that the P. cornucopiae rDNA units were derived from two kinds of ancestors (type I and II) by insertion or deletion events (100-700 bp) in the IGS 1. In four dikaryotic strains, two rDNA units of the same type (I or II) differing only by the IGS 1 length, were found in a similar number of copies, and presented a meiotic segregation in homokaryotic progeny. In one progeny, some homokaryotic strains possessed two different rDNA units: one with a high copy number and another with a lower one, showing that two different rDNA units could coexist in a single nucleus. [ABSTRACT FROM AUTHOR]

The variability of the sexual incompatibility genes of Agrocybe aegerita was investigated in the homokaryotic progeny of 13 wild dikaryotic strains from five distinct European geographic origins. Results of mating tests allowed identification of 18 A alleles and 16 B alleles out of a possible 26 different alleles for each in the sample. The determination and the comparison by a contingency χ test of the frequencies of allele replications between intra- and interregional matings showed no departure from a random distribution of incompatibility alleles. The allelic series estimated for the incompatibility genes of the entire population of A. aegerita, 30 A and 25 B aleles, are significantly less extensive than those already hypothesized for other tetrapolar hymenomycetes. However, the low variability of incompatibility genes has little effect on the outbreeding efficiency (92.6%) of this mushroom. The low variability of the incompatibility alleles and the apparent absence of intrafactorial recombination could relate to a single-locus structure of the incompatibility genes in A. aegerita. [ABSTRACT FROM AUTHOR]