A new methodology combining PCR, cloning, and sequencing of clones discriminated by RFLP for the study of microbial populations: application to an UASB reactor sample.

This work describes a methodology combining DNA extraction, polymerase chain reaction amplification with primers targeting 16S ribosomal RNA genes, cloning, and sequencing of clones previously analyzed by restriction fragment length polymorphism (RFLP), which can be applied to study the microbial diversity in a given habitat. The methodology allows the minimization of the sequencing effort, which is particularly relevant when analyzing large numbers of clones. The methodology does not require particularly skilled personnel and can easily be adaptable to the molecular characterization of virtually any particular microbial population, provided that both adequate primers and suitable restriction enzymes for RFLP analysis of the clone library have been chosen.

Aortobronchopulmonary fistula in the postoperative period of aortic coarctation.

Background And Objectives: In patients with hemoptysis and a history of aortic surgery, the possibility of aortobronchopulmonary fistula must always be considered. The objective of this study was to report a rare case of hemoptysis due to aortobronchopulmonary fistula in the late postoperative period of aortic surgery.

Case Report: Female patient, 34 years, surgical correction of aortic coarctation, presenting massive hemoptysis.

Urinary porphyrin excretion in children with mercury amalgam treatment: findings from the Casa Pia Children's Dental Amalgam Trial.

Increases in the urinary concentrations of pentacarboxyl- and coproporphyrins and the appearance of the atypical precoproporphyrin have been defined in relation to mercury (Hg) body burden in animal studies, and this change in the porphyrin excretion pattern has been described as a biomarker of occupational Hg exposure and toxicity in adult human subjects. In the present studies, urinary porphyrins were determined in relation to Hg exposure in children and adolescents, 8-18 yr of age, over the 7-yr course of a clinical trial designed to evaluate the neurobehavioral and renal effects of dental amalgam in children. Subjects were randomized to either dental amalgam or composite resin treatments.

Urinary porphyrin excretion in normal children and adolescents.

Background: Urinary porphyrins are diagnostic of various metabolic disorders and xenobiotic exposures, but comprehensive normative data for urinary porphyrin concentrations in children are currently unavailable.

Methods: Subjects were participants in a prospective, randomized, controlled clinical trial of dental materials safety, 8 to 12 y at inception, who were followed longitudinally for 7 y after baseline with an extensive battery of neurobehavioral, neurological, renal function and urinary porphyrin assessments. Porphyrins were quantified by HPLC.

Traumatic bilateral atlantoaxial rotatory subluxation mimicking as torticollis in an adult female.

Rotatory subluxation of the atlantoaxial joint in adults is a rare, but potentially life-threatening injury. Delayed or missed diagnosis can lead to catastrophic consequences. Early recognition and prompt treatment is paramount to avoid long-term morbidity and disability.

Multiway chemometric decomposition of EEM of fluorescence of CdTe quantum dots obtained as function of pH.

The effect of the pH (from 3 to 10) on the excitation emission matrices (EEMs) of fluorescence of CdTe quantum dots (QDs), capped with mercaptopropionic acid (MPA), were analyzed by multiway decomposition methods of parallel factor analysis (PARAFAC), a variant of the parallel factor analysis method (PARAFAC2) and multivariate curve resolution alternating least squares (MCR-ALS). Three different sized CdTe QDs with emission maximum at 555 nm (QDa), 594 nm (QDb) and 628 nm (QDc) were selected for analysis. The three-way data structures composed of sets of EEMs obtained as function of the pH (EEMs, pH) do not have a trilinear structure.

Burkholderia cenocepacia J2315 acyl carrier protein: a potential target for antimicrobials' development?

This work describes the isolation and characterization of an acyl carrier protein (ACP) mutant from Burkholderia cenocepacia J2315, a strain of the Burkholderia cepacia complex (Bcc). Bcc comprises at least 9 species that emerged as opportunistic pathogens able to cause life-threatening infections, particularly severe among cystic fibrosis patients. Bacterial ACPs are the donors of the acyl moiety involved in the biosynthesis of fatty acids, which play a central role in metabolism.

Dental hygiene work in a clinical trial.

Unlabelled: Dental hygiene activities were developed as part of a randomized clinical trial designed to assess the safety of low-level mercury exposure from dental amalgam restorations. Along with dental-hygiene clinical work, a community programme was implemented after investigators noticed the poor oral hygiene habits of participants, and the need for urgent action to minimize oral health problems in the study population.

Objectives: Clinical and community activity goal was to promote oral health and prevent new disease.

A longitudinal study of neurological soft signs from late childhood into early adulthood.

Neurological examination of children includes the screening for soft neurological signs (NSS). There is little knowledge about their evolution during adolescence, except that their lasting presence has been associated with developmental, psychological, and cognitive disorders. We report the results of a NSS exam (assessing gross and fine motor function and the presence of hyperactivity and motor impersistence) over a 5-year period, among a group of healthy children who were followed annually as part of a dental study.

Biomarkers of kidney integrity in children and adolescents with dental amalgam mercury exposure: findings from the Casa Pia children's amalgam trial.

Mercury is toxic to the kidney, and dental amalgam is a source of mercury exposure. Few studies have evaluated the effects of dental amalgam on kidney function in a longitudinal context in children. Here, we evaluated urinary concentrations of glutathione S-transferases (GSTs) alpha and pi as biomarkers of renal proximal and distal tubular integrity, respectively, and albumin as a biomarker of glomerular integrity in children and adolescents 8-18 years of age over a 7-year course of dental amalgam treatment.

Functional analysis of the Burkholderia cenocepacia J2315 BceAJ protein with phosphomannose isomerase and GDP-D-mannose pyrophosphorylase activities.

The bceA(J) gene from the cystic fibrosis isolate Burkholderia cenocepacia J2315 encodes a 56-kDa bifunctional protein, with phosphomannose isomerase (PMI) and guanosine diphosphate (GDP)-mannose pyrophosphorylase (GMP) activities, a new member of the poorly characterised type II PMI class of proteins. Due to the lack of homology between the type II PMIs and the human PMI, this class of proteins are being regarded as interesting potential targets to develop new antimicrobials. The BceA(J) protein conserves the four typical motifs of type II PMIs: the pyrophosphorylase signature, the GMP active site, the PMI active site and the zinc-binding motif.

Variation of the antimicrobial susceptibility profiles of Burkholderia cepacia complex clonal isolates obtained from chronically infected cystic fibrosis patients: a five-year survey in the major Portuguese treatment center.

The treatment of cystic fibrosis (CF) patients chronically infected with Burkholderia cepacia complex (Bcc) bacteria requires extensive and aggressive antibiotics therapy, exposing these bacteria to prolonged antibiotics-selective pressure. In the present study, we have compared the susceptibility patterns to 13 antimicrobials of 94 Bcc isolates obtained from 15 Portuguese CF patients in the course of chronic infection during a five-year survey. These isolates were previously genotyped and represent 11 different strains of the species B.

Dental amalgam and antibiotic- and/or mercury-resistant bacteria.

Mercury emitted from dental amalgam may select for increased numbers of antibiotic- or mercury-resistant commensal bacteria in patients and increase their risk for bacterial diseases that are resistant to common therapies. We hypothesized that the presence of dental amalgams would increase the level of mercury-, tetracycline-, ampicillin-, erythromycin-, or chloramphenicol-resistant oral and urinary bacteria as compared with levels in children receiving composite fillings. Samples were collected at baseline, 3-6 months after the initial dental treatment, and annually for 7 years of follow-up.

Optimization of Verapamil drug analysis by excitation-emission fluorescence in combination with second-order multivariate calibration.

Excitation emission fluorescence matrices (EEMs) of Verapamil drug were obtained by direct and by derivatization fluorescence spectroscopy. The fluorescence excitation and emission wavelengths were displaced to longer wavelengths and the fluorescence intensity was enhanced upon derivation with respect to the native fluorescence of the drug. The complete EEM of the native fluorescence of the drug and of the derivatization product were rapidly acquired by using a charged-coupled device detector (CCD), which is advantageous in terms of speed in the analysis, with respect to the use of a conventional photomultiplier detector.

Absolute phase image reconstruction: a stochastic nonlinear filtering approach.

This paper formulates and proposes solutions to the problem of estimating/reconstructing the absolute (not simply modulo-2pi) phase of a complex random field from noisy observations of its real and imaginary parts. This problem is representative of a class of important imaging techniques such as interferometric synthetic aperture radar, optical interferometry, magnetic resonance imaging, and diffraction tomography. We follow a Bayesian approach; then, not only a probabilistic model of the observation mechanism, but also prior knowledge concerning the (phase) image to be reconstructed, are needed.

Unsupervised contour representation and estimation using B-splines and a minimum description length criterion.

This paper describes a new approach to adaptive estimation of parametric deformable contours based on B-spline representations. The problem is formulated in a statistical framework with the likelihood function being derived from a region-based image model. The parameters of the image model, the contour parameters, and the B-spline parameterization order (i.

Neurological outcomes in children with and without amalgam-related mercury exposure: seven years of longitudinal observations in a randomized trial.

Background: Although large-scale, randomized trials involving children have been completed and their results demonstrate an absence of neurobehavioral effects from clinical exposure to mercury amalgam, neurological findings from such studies have not been reported.

Methods: The authors conducted a randomized, prospective trial examining the safety of dental amalgam in which 507 children aged 8 through 12 years were assigned to treatment with either amalgam or resin-based composite. During seven years of follow-up, the authors performed annual clinical neurological examinations, including an evaluation of neurological hard signs (NHSs), presence of tremor and neurological soft signs (NSSs).

The ZpiM algorithm: a method for interferometric image reconstruction in SAR/SAS.

This paper presents an effective algorithm for absolute phase (not simply modulo-2-pi) estimation from incomplete, noisy and modulo-2pi observations in interferometric aperture radar and sonar (InSAR/InSAS). The adopted framework is also representative of other applications such as optical interferometry, magnetic resonance imaging and diffraction tomography. The Bayesian viewpoint is adopted; the observation density is 2-pi-periodic and accounts for the interferometric pair decorrelation and system noise; the a priori probability of the absolute phase is modeled by a compound Gauss-Markov random field (CGMRF) tailored to piecewise smooth absolute phase images.

Factorial analysis optimization of a Diltiazem kinetic spectrophotometric quantification method.

A Diltiazem kinetic spectrophotometric method was optimized by factorial analysis. The experimental method is based on a two-stage reaction of Diltiazem with hydroxylamine and a ferric salt: in the first stage there is a hydroxamic acid formation; and, in the second stage there is a red colour complex ferric hydroxamate formation. The variables under investigation were: solvent; hydroxylamine, sodium hydroxide and ammonium ferric sulphate concentrations; volume of perchloric acid; and, temperature.

Evaluation of a 7-year school-based community dental hygiene programme in Portugal by high school teachers.

Objectives: To obtain high school teachers evaluation of a Community Dental Hygiene programme, developed as part of a clinical trial designed to assess the safety of low-level mercury exposure from amalgam restorations.

Methods: A questionnaire to assess programme evaluation, personal opinion on programme relevance and satisfaction with activities was distributed among teachers. It had a total of 22 questions organized into three groups.

Longitudinal urinary creatinine excretion values among preadolescents and adolescents.

To present longitudinal urinary creatinine excretion data for developmentally normal children 9-17 years of age. Only extremely limited data have been presented of a longitudinal nature for this age group. Overall, 507 children who participated in a prospective, randomized, controlled trial of dental materials safety were followed longitudinally for 7 years with renal measures, including creatinine excretion.

The contribution of dental amalgam to urinary mercury excretion in children.

Background: Urinary mercury concentrations are widely used as a measure of mercury exposure from dental amalgam fillings. No studies have evaluated the relationship of these measures in a longitudinal context in children.

Objective: We evaluated urinary mercury in children 8-18 years of age in relation to number of amalgam surfaces and time since placement over a 7-year course of amalgam treatment.

On the structural diversity of sialoliths.

Sialoliths from parotid and submaxillar glands have been characterized. Fractured and polished surfaces revealed an intrinsic structural diversity across the calculi sections. In general, the calculi presented highly mineralized amorphous-looking cores surrounded by concentric alternating mineralized and organic layers.

Virulence of Burkholderia cepacia complex strains in gp91phox-/- mice.

In cystic fibrosis (CF), infection with Burkholderia cepacia complex (Bcc) strains may cause long-term asymptomatic airway colonization, or severe lung infection leading to rapid pulmonary decline. To assess the virulence of Bcc strains, we established a lung infection model in mice with a null allele of the gene involved in X-linked chronic granulomatous disease (CGD). CGD mice, challenged intratracheally with 10(3) cells of the epidemic Burkholderia cenocepacia strain J2315, died within 3 days from sepsis after bacteria had multiplied to 3.

Survival and reasons for failure of amalgam versus composite posterior restorations placed in a randomized clinical trial.

Background: Failure of dental restorations is a major concern in dental practice. Replacement of failed restorations constitutes the majority of operative work. Clinicians should be aware of the longevity of, and likely reasons for the failure of, direct posterior restorations.