Nucleotide sequence and mutational analysis of an immunity repressor gene from Bacillus subtilis temperate phage ϕ105

We have identified and sequenced a bacteriophage phi 105 gene encoding an immunity repressor, the first to be characterized from a temperate phage infecting a Gram-positive host. Using superinfection immunity as an assay for repressor function, the phi 105 repressor gene was located within a 740-bp PvuII-HindIII subfragment near the left end of the phi 105 EcoRI-F fragment. We show that the repressor is specified by the 5'-proximal coding sequence of a translationally overlapping gene pair, transcribed from right to left on the conventional phi 105 map. Comparison of its amino acid sequence (146 residues) with that of a large number of Gram-negative bacterial and phage repressors revealed a putative DNA-binding region between positions 20 and 39. The coding region is preceded by a strong Shine-Dalgarno sequence 5' AAAGGAG 3'. Deletion analysis of the 5'-flanking DNA allowed to identify transcriptional control elements. Their structure, 5' TTGTAT 3' at -35 and 5' TATAAT 3' at -10, strongly suggests that the phi 105 repressor gene is transcribed by the major vegetative form of B. subtilis RNA polymerase, as would be expected for an early phage gene

Gouty arthritis of the spine in a renal transplant patient : a clinical case report: an unusual presentation of a common disease

Axial gout is a well-documented but uncommon manifestation of gout. Its mimicking nature and the impracticality of axial joint aspiration might considerably delay its diagnosis. We report a case in a normouricemic renal transplant recipient, whereby the primary symptom of severe neck pain suggested pyogenic spondylodiscitis as an initial tentative diagnosis. Clinical findings included a high C-reactive protein concentration and elevated body temperature. The patient did not respond to empiric antibiotic treatment and suffered consecutive attacks of severe wrist and ankle pain in conjunction with a persistent fever. Blood and joint cultures were negative, but analysis of aspirated ankle joint fluid revealed monosodium urate crystals. A dual-energy computed tomography scan confirmed the presence of monosodium urate crystals in the costovertebral joints. Colchicine treatment dramatically improved the patient's clinical condition. Axial gout should be considered in transplant recipients with severe neck or back pain, fever, and increased inflammatory parameters with a high likelihood of an infectious etiology, despite the presence of paradoxically normal or even decreased serum urate concentrations. Dual-energy computed tomography is a noninvasive technique of possible benefit in the detection of axial gout when joint fluid aspiration is not deemed safe

High-efficiency transformation of Agrobacterium tumefaciens with plasmid DNA by electroporation

We describe a very simple and highly efficient procedure for direct transformation of Agrobacterium tumefaciens with plasmid DNA. The protocol is based on high-voltage electropermeabilization of whole cells, and reproducible yields of 107 to 5 x 108 transformants per µg of plasmid DNA are obtained. basically electroporation is performed in 15% polyethylene glycol using an electrical field strength of 12.5 kV/cm. The utility of this transformation method is demonstrated by direct establishment in Agrobacterium tumefaciens of a representative genomic library from the plant Arabidopsis thaliana. Our results offer interesting prospects for the transfer of gene banks and for gene complementation in plants without the use of Escherichia coli as an intermediate host

CHARACTERIZATION OF THE BACILLUS-STEAROTHERMOPHILUS MANGANESE SUPEROXIDE-DISMUTASE GENE AND ITS ABILITY TO COMPLEMENT COPPER-ZINC SUPEROXIDE-DISMUTASE DEFICIENCY IN SACCHAROMYCES-CEREVISIAE

Recombinant clones containing the manganese superoxide dismutase (MnSOD) gene of Bacillus stearothermophilus were isolated with an oligonucleotide probe designed to match a part of the previously determined amino acid sequence. Complementation analyses, performed by introducing each plasmid into a superoxide dismutase-deficient mutant of Escherichia coli, allowed us to define the region of DNA which encodes the MnSOD structural gene and to identify a promoter region immediately upstream from the gene. These data were subsequently confirmed by DNA sequencing. Since MnSOD is normally restricted to the mitochondria in eucaryotes, we were interested (i) in determining whether B. stearothermophilus MnSOD could function in eucaryotic cytosol and (ii) in determining whether MnSOD could replace the structurally unrelated copper/zinc superoxide dismutase (Cu/ZnSOD) which is normally found there. To test this, the sequence encoding bacterial MnSOD was cloned into a yeast expression vector and subsequently introduced into a Cu/ZnSOD-deficient mutant of the yeast Saccharomyces cerevisiae. Functional expression of the protein was demonstrated, and complementation tests revealed that the protein was able to provide tolerance at wild-type levels to conditions which are normally restrictive for this mutant. Thus, in spite of the evolutionary unrelatedness of these two enzymes, Cu/ZnSOD can be functionally replaced by MnSOD in yeast cytosol

Early target attainment of continuous infusion piperacillin/tazobactam and meropenem in critically ill patients : a prospective observational study

To evaluate target attainment of empirically dosed continuous infusion piperacillin/tazobactam (TZP) and meropenem (MER) in critically ill patients.Patients were sampled on a daily basis. TZP or MER concentrations were evaluated during the first two days antibiotic therapy. The lower limit of the target range was defined as unbound concentrations equaling 4 times the epidemiological cutoff value of P. aeruginosa. The upper limit of the target range was based on the risk of toxicity, i.e. unbound concentrations >160 mg/L for TZP and > 45 mg/L for MER. Multivariable logistic regression was used to evaluate factors associated with target attainment.Data from 253 patients were analyzed. Overall, 76/205 (37.1%) and 36/48 (75%) of the patients receiving TZP or MER respectively, attained target concentrations. In multivariable analysis, estimated creatinine clearance was identified as a risk factor for target non-attainment (OR 0.988, 95%CI [0.982;0.994]). Patients receiving MER were more likely to attain target concentrations compared with patients receiving TZP (OR 6.02, 95%CI [2.12;18.4]).Target attainment of empiric antibiotic therapy in critically ill patients was low (37%) for TZP and moderate (75%) for MER, despite the use of a loading dose and despite optimization of the mode of infusion

Non-rigid registration of serial intra-operative images for automatic brain shift estimation

Measurement of intra-operative brain motion is important to provide boundary conditions to physics-based deformation models that can be used to register pre- and intra-operative information. In this paper we present and test a technique that can be used to measure brain surface motion automatically. This method relies on a tracked laser range scanner (LRS) that can acquire simultaneously a picture and the 3D physical coordinates of objects within its field of view. This reduces the 3D tracking problem to a 2D non-rigid registration problem which we solve with a Mutual Information-based algorithm. Results obtained on images of a phantom and on images acquired intra-operatively that demonstrate the feasibility of the method are presented

Identification of Extracellular d-Catenin Accumulation for Prostate Cancer Detection

BACKGROUND—Prostate cancer is the second leading cause of cancer death in men, and early detection is essential to reduce mortality and increase survival. δ-Catenin is a unique β-catenin superfamily protein primarily expressed in the brain but is upregulated in human prostatic adenocarcinomas. Despite its close correlation with the disease, it is unclear whether δ-catenin presents the potential in prostate cancer screening because it is an intracellular protein. In this study, we investigated the hypothesis of δ-catenin accumulation in the urine of prostate cancer patients and its potential pathways of excretion into extracellular milieu. METHODS—Prostate cancer cell cultures, human tissue biopsies, and voided urines were characterized to determine extracellular δ-catenin accumulation and co-isolation with exosomes/ prostasomes. RESULTS—We identified δ-catenin in culture media and in the stroma of human prostate cancer tissues. In PC-3 cells in culture, δ-catenin was partially co-localized and co-isolated with raftassociated membrane protein caveolin-1 and glycosylphosphatidylinositol-anchored protein CD59, suggesting its potential excretion into extracellular milieu through exosome/prostasome associated pathways. Interference with endocytic pathway using wortmannin did not block prostasome excretion, but δ-catenin overexpression promoted the extracellular accumulation of caveolin-1. δ- Catenin, caveolin-1, and CD59 were all detected in cell-free human voided urine prostasomes. δ- Catenin immunoreactivity was significantly increased in the urine of prostate cancer patients (p<0.0005). CONCLUSIONS—This study demonstrated, for the first time, the extracellular accumulation of δ-catenin in urine supporting its potential utility for non-invasive prostate cancer detection. Originally published The Prostate, Vol. 69, No. 4, March 1 200

Studentenparticipatie als een strategie om leiderschapskwaliteiten te ontwikkelen

België is een van de weinige landen ter wereld waar de graad van studentenparticipatie in de opleiding geneeskunde zo hoog ligt. In deze studie gaan we na in hoeverre de studentenparticipatie kan bijdragen aan het ontwikkelen van leiderskwaliteiten die van belang zijn voor een arts. Studentenvertegenwoordigers uit de opleiding geneeskunde aan de Vlaamse universiteiten werden een vragenlijst voorgelegd. Uit de vragenlijst blijkt dat bepaalde competenties aangeleerd worden dankzij het engagement van de student. Deze vaardigheden kunnen bijdragen tot een goede ontwikkeling van een arts die kan inspelen op de noden van de maatschappij

Saturable elimination of piperacillin in critically ill patients:implications for continuous infusion

The study aimed to evaluate saturation of piperacillin elimination in critically ill adult patients. Seventeen critically ill adult patients received continuous and intermittent infusion of piperacillin/tazobactam. Piperacillin plasma concentrations (n = 217) were analysed using population pharmacokinetic (PopPK) modelling. Post-hoc simulations were performed to evaluate the type I error rate associated with the study. Unseen data were used to validate the final model. The mean error (ME) and root mean square error (RMSE) were calculated as a measure of bias and imprecision, respectively. A PopPK model with parallel linear and non-linear elimination best fitted the data. The median and 95% confidence interval (CI) for the model parameters drug clearance (CL), volume of central compartment (V), volume of peripheral compartment (V-p) and intercompartmental clearance (Q) were 9 (7.69-11) L/h, 6.18 (4.93-11.2) L, 11.17 (7.26-12) L and 15.61 (12.66-23.8) L/h, respectively. The Michaelis-Menten constant (K-m) and the maximum elimination rate for Michaelis-Menten elimination (V-max) were estimated without population variability in the model to avoid overfitting and inflation of the type I error rate. The population estimates for K-m and V-max were 37.09 mg/L and 353.57 mg/h, respectively. The bias (ME) was -20.8 (95% CI -26.2 to -15.4) mg/L, whilst imprecision (RMSE) was 49.2 (95% CI 41.2-56) mg/L. In conclusion, piperacillin elimination is (partially) saturable. Moreover, the population estimate for K-m lies within the therapeutic window and therefore saturation of elimination should be accounted for when defining optimum dosing regimens for piperacillin in critically ill patients. (C) 2019 Elsevier B.V. and International Society of Chemotherapy. All rights reserved