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[Mycobacterium fortuitum skin infection occurring after a facelift].

INTRODUCTION: Mycobacterium fortuitum skin infections are rare and usually iatrogenic. We report a case with cervical involvement following a facelift. OBSERVATION: A 65 year-old woman, without past history, underwent bilateral surgical facelift, complicated by cutaneous necrosis and treated with directed healing at home. Six weeks later, an abscessed nodule appeared under the left maxillary and was drained surgically. Then other pre-auricular and left cervical inflammatory nodules appeared without adenopathy or fever. M. fortuitum was isolated in bacteriological samples. The initially probabilistic antibiotherapy with carithromycin, subsequently adapted with amikacine and cirprofloxacine and then imipeneme for a total duration of 3 months, led to the clinical cure. DISCUSSION: Mycobacterium fortuitum is a rapidly growing, ubiquitous, mycobacteria responsible for nosocomial infections in immunocompetent patients, notably following plastic surgery. Contamination occurs where there has been a rupture in the skin barrier through contact with a vector (water, surgical material, antiseptic.). Treatment, which is not codified, consists in the association of surgery and antibiotics for several months.

Aged↗

High-performance liquid chromatographic validated assay of doxorubicin in rat plasma and tissues.

A specific and selective high-performance liquid chromatography (HPLC) technique that requires few manipulations, and is readily adaptable to analysis for a large series of samples, has been developed for the determination of the concentration of the anticancer drug doxorubicin (DXR) in rat serum and tissues. The biological samples were efficiently deproteinised and resolved from a reversed-phase nucleosil C18 column with fluorescence detection. The validation study of the proposed method was successfully carried out in an assay range of between 5 and 5000 ng/ml and was subsequently implemented in a pharmacokinetic study of DXR in Wistar rats that were treated by intravenous administration of the drug.

Animals↗

[Perioperative perfusion in children: evaluation of a new perfusion solution].

A new intravenous solution (B66) containing 0.9% dextrose in water for infusion therapy in infants and children was assessed. Forty-one children, aged between 6 months and 11 years, scheduled for elective non haemorrhagic surgery, were randomly assigned to two groups: children in group I (n = 22) were given 1% dextrose in lactated Ringer's solution (RLG1), and those in group II (n = 19) the commercially available solution B66 (0.9% dextrose in lactated Ringer's solution). The fluids were administered throughout the study with volumetric infusion pumps (IVAC 541). The infusion rate was adapted to children's weight and age. Blood samples for blood glucose, sodium and protein concentration assessments were obtained at induction (T0), on arrival in the recovery room (T1), than 30 and 60 min later (T2 and T3). Preoperative blood glucose concentrations were within the normal range for all children except for two, who had asymptomatic hypoglycaemia (2.2 and 2.3 mmol.l-1). Postoperative blood glucose concentrations were higher in both groups. This increase was significantly greater in the RLG1 group than in the B66 group. The highest mean figures at T1 were 6.8 +/- 1.5 mmol.l-1 and 5.2 +/- 1.0 mmol.l-1 in the RLG1 and B66 groups respectively. Total protein levels decreased postoperatively significantly in both groups. Preoperative age-related differences in total protein concentrations were also observed postoperatively. Sodium concentrations remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Glucose↗

Extraction and quantitative analysis of iodine in solid and solution matrixes.

129I is a contaminant of interest in the vadose zone and groundwater at numerous federal and privately owned facilities. Several techniques have been utilized to extract iodine from solid matrixes; however, all of them rely on two fundamental approaches: liquid extraction or chemical/heat-facilitated volatilization. While these methods are typically chosen for their ease of implementation, they do not totally dissolve the solid. We defined a method that produces complete solid dissolution and conducted laboratory tests to assess its efficacy to extract iodine from solid matrixes. Testing consisted of potassium nitrate/potassium hydroxide fusion of the sample, followed by sample dissolution in a mixture of sulfuric acid and sodium bisulfite. The fusion extraction method resulted in complete sample dissolution of all solid matrixes tested. Quantitative analysis of 127I and 129I via inductively coupled plasma mass spectrometry showed better than +/-10% accuracy for certified reference standards, with the linear operating range extending more than 3 orders of magnitude (0.005-5 microg/L). Extraction and analysis of four replicates of standard reference material containing 5 microg/g 127I resulted in an average recovery of 98% with a relative deviation of 6%. This simple and cost-effective technique can be applied to solid samples of varying matrixes with little or no adaptation.

Journal Article↗

(H)N(COCA)NH and HN(COCA)NH experiments for 1H-15N backbone assignments in 13C/15N-labeled proteins.

Triple resonance HN(COCA)NH pulse sequences for correlating 1H(i), 15N(i), 1H(i-1), and 15N(i-1) spins that utilize overlapping coherence transfer periods provide increased sensitivity relative to pulse sequences that utilize sequential coherence transfer periods. Although the overlapping sequence elements reduce the overall duration of the pulse sequences, the principal benefit derives from a reduction in the number of 180 degrees pulses. Two versions of the technique are presented: a 3D (H)N(COCA)NH experiment that correlates 15N(i), 1H(i-1), and 15N(i-1) spins, and a 3D HN(COCA)NH experiment that correlates 1H(i), 15N(i), 1H(i-1), and 15N(i-1) spins by simultaneously encoding the 1H(i) and 15N(i) chemical shifts during the t1 evolution period. The methods are demonstrated on a 13C/15N-enriched sample of the protein ubiquitin and are easily adapted for application to 2H/13C/15N-enriched proteins.

Carbon Isotopes↗

Validation of an oral health quality of life index (GOHAI) in France.

UNLABELLED: Oral health has traditionally been defined in terms of disease. Today, health is seen in a wider context: taking into account its impact on everyday living. Several indices attempt to capture this dimension. The Geriatric Oral Health Assessment Index (GOHAI) has been adapted for general use and tested on adult samples, mainly in North America. Language, cultural norms and the health care system in France are different and this raises the need to validate the GOHAI in France before it receives widespread use. OBJECTIVES: The purpose of this study was to test the validity of a French version of the GOHAI. METHODS: The GOHAI is based on responses to a 12-item self-administered questionnaire. The items were translated into French, back-translated and compared with the original. After pilot testing and minor modifications, the French version was administered to a group of low-income persons benefiting from the national health insurance system (n = 260, 18-45 years). Measures for stability and internal consistency were calculated. Concurrent and discriminant validity were assessed. RESULTS: Cronbach's alpha (0.86) showed a high internal consistency and homogeneity between items. Item-scale correlations varied between 0.40 and 0.78. Repeat administration of the GOHAI to 32 participants gave weighted kappa coefficients from 0.51 to 0.87 and a Pearson's correlation coefficient of 0.87. Low GOHAI scores were associated with perceptions of poor oral and general health, low satisfaction with oral health and a perceived need for dental care. There were significant relationships between the GOHAI score and most objective measures of dental status except FT. Younger, well-educated and higher income respondents were more likely to have a high GOHAI score. CONCLUSION: The French version of the GOHAI exhibits satisfactory psychometric properties but two items (one about swallowing, the other with complex sentence structure) had poor stability.

Adolescent↗

Monitoring of CD34+ cells during leukapheresis allows a single, successful collection of hemopoietic progenitors in patients with low numbers of circulating stem cells.

We have studied a total of 188 patients with hematological malignancies, submitted to mobilization therapy with G-CSF associated or not with chemotherapy in order to: (1) establish the lower limit of circulating progenitor cells that allows the collection of 2 x 10(6) CD34+ cells/kg by a single leukapheresis, utilizing the instrument set on standard parameters; (2) evaluate whether the number and quality of CD34+ cells collected remain stable during leukapheresis; and (3) collect a sufficient number of circulating CD34+ cells by a single procedure in patients in whom such an approach would have been insufficient to reach the target with the instrument set on standard parameters. The retrospective analysis conducted in 85 patients showed that 19 circulating CD34+ cells/microl represented the cut-off number capable of discriminating between patients who will require one or more apheresis to collect 2 x 10(6) CD34+ cells/kg. The validity of this value was prospectively confirmed in 70 subsequent patients. Based on in vitro results that showed the stability in the number of CD34+ cells, the proportion of different CD34+ cell subpopulations and the clonogenic capacity of the stem cell compartment during leukapheresis both in the blood of the patients and in samples taken directly from the instrument, we have adapted the blood volume to be processed in 33 patients with <19 PB CD34+ cells/microl. Stem cell collection was monitored during the leukapheresis and the procedure was prolonged for a time period estimated to be sufficient to reach the target number of CD34+ cells with a single procedure. The median increment of the total blood volume processed, calculated from the volume set automatically by the instrument was 25.2%, with a median of 3.3-fold total blood volume processed. In all cases, a sufficient CD34+ cell collection was completed in a single procedure. After autograft, the pattern of blood reconstitution was similar to that of all the other patients.

Adolescent↗

Generation of larger numbers of separated microbial populations by cultivation in segmented-flow microdevices.

The high speed production of fluid segments for the highly parallelized cultivation of monoclonal cell populations was carried out by the use of microchip segmentor modules. Aqueous fluid segments, embedded in a non-miscible carrier liquid, were produced with frequencies up to 30 s(-1) and showed a high homogeneity in size. This corresponds with the production of about 2.5 million samples per day. The segment volumes can be adapted between about 4 nl and 100 nl. The typical segment size for cultivation experiments is in the range between 40 nl and 80 nl. Nutrient medium can be applied instead of pure water. It is possible to aliquot a cell suspension in such a way that most of the aqueous fluid segments contain only one cell. In model experiments with four microbial species chip-produced aliquots of 60 nl, each containing one or a few cells, were incubated in Teflon capillary tubes. Rapid growth of the microcultures was observed. Cell densities were found to be as high as in conventional shake flask cultures.

Colony Count, Microbial↗

Deaf adults without attention deficit hyperactivity disorder display reduced perceptual sensitivity and elevated impulsivity on the Test of Variables of Attention (T.O.V.A.).

The Test of Variables of Attention (T.O.V.A.; R. A. Leark, T. R. Dupuy, L. M. Greenberg, C. L. Corman, & C. L. Kindeschi, 1996) is a continuous performance test used widely to help diagnose attention deficit hyperactivity disorder (ADHD) in both hearing and deaf people. The T.O.V.A. previously has been normed only on the hearing population. The T.O.V.A. performance of 38 prelingually and severely-to-profoundly deaf young adults and 34 hearing young adults who did not have ADHD was examined in this study. Deaf and hearing participants did not differ on the T.O.V.A. omission variables. However, deaf participants had significantly lower d' scores than hearing participants, indicating reduced perceptual sensitivity to the distinction between target and distractor stimuli. Consistent with the existing literature on attentional reorganization in the deaf population, this result was interpreted as indicating a deafness-related reduction in attention to centrally presented stimuli. Deaf participants also showed 2 to 3 times more commission errors than hearing participants and displayed a higher incidence of anticipatory errors. These results suggest a deafness-related increase in impulsivity at the time of response initiation. Beta score analysis confirmed that deaf participants adopted an overall less conservative (more impulsive) response criterion that contributed to their total elevated commission errors. However, a portion of the commission errors was secondary to their reduced d', not to increased behavioral impulsivity. Separate factor analyses of the standard T.O.V.A. variables revealed highly similar factor structures for deaf and hearing participants, indicating similar construct validity of the T.O.V.A. for both groups. The evidence for increased inattention and impulsivity in a non-ADHD deaf sample are interpreted in the context of an adaptive attentional reorganization due to deafness. Along with the factor analytic results, these considerations suggest that separate T.O.V.A. norms must be developed for the deaf population to avoid overdiagnosis of ADHD in deaf individuals.

Adult↗

Applied technique for increasing calicivirus detection in shellfish extracts.

AIMS: Optimal detection of enteric RNA viruses in clinical, environmental, and food products using reverse transcription-PCR (RT-PCR) when inhibitory substances in extracted sample materials are present. METHODS AND RESULTS: We adapted a device for detection of RNA viruses in plant tissues and insects to detect a calicivirus strain (San Miguel sea lion virus, serotype 17) in water and oyster tissue extracts. This single, compartmentalized tube-within-a-tube (TWT) device for RT-PCR-nested PCR was compared to a conventional protocol of RT-PCR-nested PCR. In the presence of 100 mg of shellfish tissue extract equivalent, this TWT device decreases the calicivirus assay detection limit 10-fold over that of conventional RT-PCR-nested PCR while maintaining an identical detection limit of viral nucleic acid suspended in water. Both the conventional and TWT methods estimated the total particle-to-infectious particle ratio for this strain of calicivirus at approximately 40 : 1. CONCLUSIONS: We believe that the TWT device with appropriate RT-PCR primers will decrease the detection limit for other calicivirus strains and RNA viruses in shellfish tissue extracts. SIGNIFICANCE AND IMPACT OF THE STUDY: We believe that the TWT approach is applicable to other situations where RT and/or PCR inhibitory materials are present or nucleic acid targets of bacteria or viruses are at low levels in extracts of food products or clinical specimens.

Animals↗

Variation of proteolytic activity in ewe during in vivo digestion of two pea-based diets differing by their fermentability characteristics.

In order to study the effects of a small difference in starch and nitrogen availability on proteolysis, two different diets were supplied to four ewes fitted with rumen fistulae. They differed in the ratio of fermentable nitrogen over fermentable energy. with 144 g of fermentable nitrogen (FN) per kg of fermentable energy (FE) for diet I and 126 g FN x kg(-1) FE for diet II. The diets were constituted of 700 g hay grass, 200 g ground pea and either 100 g ground wheat (diet I) or 100 g corn starch (diet II). After two weeks of an adapting period to the diets, rumen content was sampled after feeding over time. The rate of disappearance of soluble proteins was 2.5 times higher with diet II and ammonia concentrations were significantly lower (from -28 to -43%) with diet II. Total proteolytic activity, by considering all the bacterial compartments, was significantly higher with diet II (+40 EU/mL x h(-1)): changes in the total proteolytic activity in the particulate and the liquid phases of the rumen could explain the difference observed between the two diets. Moreover, with diet II, exopeptidase activities increased more in the liquid phase, especially leucine aminopeptidase and Dipeptidyl peptidase I (DPP-I), and the diversity of endopeptidase activities increased in the particulate phase. These two facts could account for the higher total proteolytic activity in the rumen content with diet II.

Animal Feed↗

Genetic exchange and plasmid transfers in Borrelia burgdorferi sensu stricto revealed by three-way genome comparisons and multilocus sequence typing.

Comparative genomics of closely related bacterial isolates is a powerful method for uncovering virulence and other important genome elements. We determined draft sequences (8-fold coverage) of the genomes of strains JD1 and N40 of Borrelia burgdorferi sensu stricto, the causative agent of Lyme disease, and we compared the predicted genes from the two genomes with those from the previously sequenced B31 genome. The three genomes are closely related and are evolutionarily approximately equidistant ( approximately 0.5% pairwise nucleotide differences on the main chromosome). We used a Poisson model of nucleotide substitution to screen for genes with elevated levels of nucleotide polymorphisms. The three-way genome comparison allowed distinction between polymorphisms introduced by mutations and those introduced by recombination using the method of phylogenetic partitioning. Tests for recombination suggested that patches of high-density nucleotide polymorphisms on the chromosome and plasmids arise by DNA exchange. The role of recombination as the main mechanism driving B. burgdorferi diversification was confirmed by multilocus sequence typing of 18 clinical isolates at 18 polymorphic loci. A strong linkage between the multilocus sequence genotypes and the major alleles of outer-surface protein C (ospC) suggested that balancing selection at ospC is a dominant force maintaining B. burgdorferi diversity in local populations. We conclude that B. burgdorferi undergoes genome-wide genetic exchange, including plasmid transfers, and previous reports of its clonality are artifacts from the use of geographically and ecological isolated samples. Frequent recombination implies a potential for rapid adaptive evolution and a possible polygenic basis of B. burgdorferi pathogenicity.

Amino Acid Sequence↗

Sexual harassment of college students: implications for campus health promotion.

The authors examined students' perceptions, attitudes, and experiences of sexual harassment by faculty members at a state university, using a sample of 1,139 graduate and undergraduate students. Twenty-six percent (292) of those in the sample responded. The instrument used in the study, adapted from a survey previously used at the University of Iowa, operationally defined eight categories of behavior: sexist comments, undue attention, verbal sexual advances, body language, invitations, physical advances, explicit sexual propositions, and sexual bribery. As many as 8% of the respondents indicated they had experienced the three most extreme forms of harassment--physical advances, explicit sexual propositions, and sexual bribery. Although most students thought they would report sexual harassment, only three incidents of the most extreme forms of sexual harassment were actually reported. Those who experienced harassment indicated that it generally came from one rather than from several faculty members and that it came from both male and female faculty. Twenty-three percent of the men reported experiencing sexist comments, and 5 male students reported they had experienced at least one of the three most extreme forms of sexual harassment. Recommendations for policy revisions and campus health promotion programming that were made following the survey are discussed.

Adult↗

Inhalant abuse and the abuse of other drugs.

AIMS: To examine the relationship between inhalant abuse and other substances of abuse. DESIGN: Survey using a structured interview administered by a single trained interviewer. SETTING: A juvenile detention facility. PARTICIPANTS: 209 children incarcerated at the facility over a 3-month period. SELECTION PROCEDURE: Consecutive sample. INTERVENTIONS: None. MEASUREMENTS/FINDINGS: The structured interview was adapted from the American Drug and Alcohol Survey, which has been extensively used to obtain substance abuse epidemiologic data. We collected information on inhalants, alcohol, marijuana, downers, pep pills, lysergic acid diethylamide (LSD), cocaine, designer drugs, phencyclidine (PCP), Talwin and Ritalin, speed, and narcotics. The chi-square or Fisher exact test were used when appropriate. Mean ages of initial experimentation were as follows: inhalants, 9.7 years; marijuana, 11.9 years; alcohol (inebriated), 12.0 years; cigarettes, 11.2 years; for the remaining substances of abuse, the mean age was 13.2-14.7 years. Thirty subjects had used inhalants. Significant relationships were found between inhalants and cocaine (p = .004), Talwin and Ritalin (p = .001), downers (p = .01), and narcotics (p = .003). CONCLUSIONS: For children incarcerated in a juvenile detention facility in our community, inhalant abuse is associated with the later use of other substances of abuse. If this finding is replicated in other populations, it underscores the need for effective preventive strategies.

Administration, Inhalation↗

Rapid detection of the staphylococcal mecA gene from BACTEC blood culture bottles by the polymerase chain reaction.

A rapid polymerase chain reaction (PCR) method for the direct detection of the staphylococcal mecA gene from BACTEC blood culture bottles (Becton Dickinson, Sparks, MD) was developed. Published primer sequences and sample preparation using Achromopeptidase for cell lysis were adapted to the use of the Idaho Technology Air Thermocycler 1605 (Idaho Technologies, Idaho Falls, ID). The method was validated with 80 strains of coagulase-positive and coagulase-negative geographically diverse methicillin-resistant and susceptible isolates of staphylococci. There was a 100% correlation between the PCR results and the results of standard susceptibility testing methods. From BACTEC 9240 blood cultures, mixed aliquots of blood and broth containing gram-positive cocci in clusters were centrifuged at low speed to sediment red blood cells. After additional centrifugation and wash steps, PCR was performed on the resuspended pellet. The turnaround time from initial Gram stain detection of positive BACTEC bottles to PCR amplicon detection by agarose gel electrophoresis is less than 3 hours. In a clinical evaluation of 181 blood culture isolates, there was a 99% correlation with standard susceptibility results for Staphylococcus aureus. Discrepant results for Staphylococcus aureus isolates were verified by a Mueller Hinton plate supplemented with 6 microg/mL of oxacillin and 2% sodium chloride. For coagulase-negative staphylococci, the PCR method detected an additional seven resistant isolates that were reported by the Vitek as susceptible. Coagulase-negative staphylococcal susceptibility results that were in disagreement with the PCR assay were confirmed by the disk-diffusion method. This procedure is accurate, rapid and fits well into laboratory work flow. Rapid detection of the mecA gene on positive blood culture vials has become a routine test in the authors' clinical microbiology laboratory.

Bacterial Proteins↗

Clinical adaptation of the sperm ubuquitin tag immunoassay (SUTI): relationship of sperm ubiquitylation with sperm quality in gradient-purified semen samples from 93 men from a general infertility clinic population.

BACKGROUND: The proteolytic chaperone peptide ubiquitin accumulates in defective human spermatozoa. Immunodetection of ubiquitin in human sperm samples correlates with semen quality and male fertility. METHODS: Semen samples from 93 men from couples seeking infertility treatment were separated on a PureSperm density gradient and screened by immunofluorescence microscopy with anti-ubiquitin antibodies. The percentage of spermatozoa with head ubiquitylation was recorded and compared with clinical semen evaluation and embryo development data after IVF or ICSI. Subjects were divided into the following four groups based on the initial clinical diagnosis of the couples; group 1, male factor; group 2, idiopathic infertility; group 3, female infertility with neither partner having children previously; and group 4, female infertility with male partners having children from previous relationships. RESULTS: The percentage of sperm with ubiquitylated heads remaining after PureSperm separation in the respective groups was 4.0% (male factor), 2.5% (idiopathic infertility), 0.7% (female infertility and presumed fertile male) and 0.9% (female infertility with established fertile male). Negative correlations between sperm ubiquitin and several parameters reflective of embryo development after assisted fertilization were found within the male factor group. CONCLUSIONS: Use of this simplified ubiquitin-based sperm quality assay is feasible in a clinical environment. Since the gradient separation does not completely deplete the defective spermatozoa, the modified light microscopic sperm ubiquitin tag immunoassay could add a new level of stringency to the selection of human spermatozoa for ICSI.

Antibodies, Monoclonal↗

Very early prenatal diagnosis of genetic diseases based on coelomic fluid analysis: a feasibility study.

BACKGROUND: Coelocentesis may represent the ideal technique for very early prenatal diagnosis. Although cell density in coelomic fluid (CF) is very low, the results of analyses on the cellular compartment have been proposed for prenatal diagnosis. METHODS AND RESULTS: We aimed to evaluate the amount of total DNA (i.e. cellular and cell-free) in 14 samples (0.4-0.8 ml) of CF, taken from women at 8- to 9-week gestation, who are about to undergo termination of pregnancy, and to assess the feasibility of multiple single-gene analyses using multiplex real-time PCR. We found that the amount of total DNA in the CF was very low and varied widely. Genetic testing using multiplex real-time PCR was successfully achieved in 10 of 14 samples (71%). However, when considering samples that could provide a reliable prenatal diagnosis (i.e. successful PCR analysis and no marked maternal contamination), reliable CF-DNA-based prenatal diagnoses were obtained in only 8 of the 14 (58%) samples. CONCLUSION: The development of highly reliable procedures adapted to pauci-cellular CF is crucially needed before coelocentesis could be proposed for early prenatal diagnosis of genetic diseases before 10 weeks.

Body Fluids↗

Effects of psychotherapy in schizophrenia: II. Comparative outcome of two forms of treatment.

This study compares the effects of exploratory, insight-oriented (EIO) and reality-adaptive, supportive (RAS) forms of psychotherapy on a sample of 95 schizophrenic patients. Analyses of 2-year outcomes revealed a complex interaction between the type of psychotherapy provided and the domain of psychopathology affected. RAS psychotherapy exerted clear preferential effects in the areas of recidivism and role performance. The EIO psychotherapy exerted preferential, albeit more modest, action in the areas of ego functioning and cognition. Overall, however, the magnitude of the differences was low. The results highlighted the need for more focused studies of subgroups, and of process and contextual influences on outcome.

Adult↗