Evolution of the environmental contamination by thermophilic fungi in a turkey confinement house in France.

Fungal species constitute a major part of environmental contaminants in facilities where animals are housed. The present investigation was aimed at describing the relative abundances of fungal species and their concentrations in a turkey confinement house in France. Fungal cultures from poultry feed, litter, and air were undertaken every week throughout the 16-wk period of breeding. The incubation temperature of 40 degrees C was selected to isolate thermophilic fungal species (especially Aspergillus spp. and Candida albicans) that are potentially pathogenic for birds. The 2 species Aspergillus fumigatus and Aspergillus flavus were recovered at a mean of 10.5 and 37.0 cfu/m(3) of air sampled, respectively. Individual samplings yielded concentrations of up to 150.0 cfu/m(3) for A. flavus in the first weeks of the investigation. Other fungal species were recovered at a mean of 18.9 cfu/m(3) (maximum 36.3 cfu/m(3)) in the air. The yeast C. albicans was first detected at wk 4 from litter samples and at wk 7 from poultry feed. Densities of C. albicans remained very high in litter samples (63.2 cfu/g) even after new litter was added at wk 10. To analyze the genetic polymorphism of A. fumigatus, the most pathogenic mold in birds, a total number of 198 isolates (134 from air, 34 from litter, and 30 from feed samples) were genotyped using 2 polymorphic microsatellite markers. More than half (42 out of 73, 57.5%) of the genotypes were detected only once. This finding suggests that the contamination of the breeding environment is not due to a single source and confirms the very high genetic diversity of environmental A. fumigatus isolates. As during the study period, no outbreak of fungal infections occurred; the levels of fungal contaminations reported here do not seem sufficient, at least alone, to trigger fungal infections.

Assessment of Helicobacter pylori gene expression within mouse and human gastric mucosae by real-time reverse transcriptase PCR.

Despite increasing knowledge on the biology of Helicobacter pylori, little is known about the expression pattern of its genome during infection. While mouse models of infection have been widely used for the screening of protective antigens, the reliability of the mouse model for gene expression analysis has not been assessed. In an attempt to address this question, we have developed a quantitative reverse transcriptase PCR (RT-PCR) that allowed the detection of minute amounts of mRNA within the gastric mucosa. The expression of four genes, 16S rRNA, ureA (encoding urease A subunit), katA (catalase), and alpA (an adhesin), was monitored during the course of a 6-month infection of mice and in biopsy samples from of 15 infected humans. We found that the selected genes were all expressed within both mouse and human infected mucosae. Moreover, the relative abundance of transcripts was the same (16S rRNA > ureA > katA > alpA), in the two models. Finally, results obtained with the mouse model suggest a negative effect of bacterial burden on the number of transcripts of each gene expressed per CFU (P < 0.05 for 16S rRNA, alpA, and katA). Overall, this study demonstrates that real-time RT-PCR is a powerful tool for the detection and quantification of H. pylori gene expression within the gastric mucosa and strongly indicates that mice experimentally infected with H. pylori provide a valuable model for the analysis of bacterial gene expression during infection.

When girls versus boys play alone: nonsocial play and adjustment in kindergarten.

The goal of the present study was to examine the relations between different forms of children's nonsocial play behaviors and adjustment in kindergarten. The participants in this study were 77 kindergarten children (38 boys, 39 girls; mean age = 66.16 months, SD = 4.11 months). Mothers completed ratings of child shyness and emotion dysregulation. Children's nonsocial play behaviors (reticent, solitary-passive. solitary active) were observed during free play. In addition, teachers rated child behavior problems (internalizing and externalizing) and social competence; academic achievement was assessed through child interviews. Results from regression analyses revealed that different types of nonsocial play were differentially associated with child characteristics and indices of adjustment. For some forms of nonsocial play, the nature of these associations differed significantly for boys and girls.

Formulations of single or multiple H. pylori antigens with DC Chol adjuvant induce protection by the systemic route in mice. Optimal prophylactic combinations are different from therapeutic ones.

The ability to induce a protective response against Helicobacter pylori infection has been investigated by systemic immunization of mice with urease formulated with the cationic lipid DC Chol. This compound acts both as a formulating agent and as an adjuvant and induces a balanced Th1/Th2 response shown to be more effective for protection in our previous studies. Urease-DC Chol induced a significant protection in prophylaxis but not in therapeutic immunization. The protection level was between 1.5 and 2 log reduction of bacterial density measured by quantitative culture compared to unimmunized-infected mice. In parallel, the protective efficacy of other H. pylori antigens formulated in a similar way and administered with DC Chol was tested. These antigens were tested alone or in combination in prophylactic and therapeutic regimens. Some combinations of antigens induced a better prophylactic or therapeutic activity than urease alone (0.5-1.5 log further reduction in prophylaxis and therapy respectively, P<0.05). The combinations that induced the best protection were different in prophylaxis and therapy. In conclusion, DC Chol provides a convenient and efficient method to formulate different antigens even when they are present in non-compatible buffers initially. Moreover, the results obtained in protection against H. pylori with such formulations should lead the way to future clinical trials.

Experimental design optimization of filamentous phage transfection into mammalian cells by cationic lipids.

A previous study showed that filamentous phage could be efficiently transfected into mammalian cells in the presence of the cationic lipid Transfectam. In the present study, we used an experimental plan based on a uniform network (Doehlert) matrix to estimate optimal transfection conditions in two different cell lines, CHO and Cos-7. Using the cationic lipid RPR120535b as a model, we show that optimal conditions can be determined much more readily than with standard response curves. Under optimal conditions as analyzed by FACS, up to 60% of Cos-7 and 50% of CHO cells can be transfected. Furthermore, a comparison of different lipids (Transfectam, RPR120535b, TC1-12 and GAP-DLRIE/DOPE) suggests that lipids with multiple amine groups are more efficient for the transfection of filamentous phage.

Modulation of apolipoprotein D and apolipoprotein E expression in rat hippocampus after entorhinal cortex lesion.

Apolipoprotein (apo) D is a member of the lipocalin family of proteins. Although its physiological function is unknown, apoD is thought to transport one or more small hydrophobic ligands. A second apolipoprotein, apoE is known to play an important role in lipid transport, and apoE genetic polymorphism has been shown to be associated with susceptibility to Alzheimer's disease. Both apoD and apoE are expressed in the central nervous system (CNS) and both proteins accumulate at sites of peripheral nerve injury due to increased local synthesis. The two proteins may have overlapping or complementary functions within nervous tissue. In order to define the role of apoD within the CNS, we have studied the regional distribution of apoD and apoE mRNA and protein within the normal rat brain and the changes in apoD and apoE expression in the hippocampus of rats after entorhinal cortex lesion (EC lesion). Within the brains of normal rats, apoD expression in the hippocampus was as high as 180-fold that of the liver. ApoD mRNA levels in other areas of the rat brain ranged from 40 to 120 times the hepatic levels. The distribution of apoE gene expression within the brain was similar to that of apoD, but was much lower than hepatic apoE expression. When rats were subjected to EC lesion, the apoD message increased by 54% at 4 days post lesion (DPL) in the ipsilateral region of hippocampus while apoE mRNA levels (ipsilateral and contralateral) decreased by 43%. At 6 to 8 DPL apoD mRNA in the ipsilateral hippocampus remained elevated (42% above controls) whereas the apoE mRNA levels increased to about 15% above those of controls. At 14 and 31 DPL, both apoD and apoE expression was similar to controls. The increase in immunoreactive apoD in hippocampal extracts was more dramatic. At 1 DPL, immunoreactive apoD levels were already 16-fold higher than those in extracts of non-lesioned animals and, at 31 DPL, levels were still 8-fold higher than those of control animals. Finally, we have demonstrated that the levels of apoD in the brains of apoE-deficient mice are 50-fold those of wildtype control mice. ApoD clearly has an important function within the CNS in both normal and pathological situations.

Cell volume regulation by the mouse zygote: mechanism of recovery from a volume increase.

Mouse zygotes regulate their volumes after cell swelling. This regulatory volume decrease (RVD) is rapid and complete. RVD in zygotes was inhibited by K+ or Cl- channel blockers, indicating the participation of such channels in volume recovery. The channels are separate entities, as indicated by the ability of the cation ionophore gramicidin to restore RVD when K+ channels are blocked but not when Cl- channels are blocked. Intracellular Ca2+ concentration increased with cell swelling. Nevertheless, RVD occurred normally in zygotes loaded with the Ca2+ chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, which prevented Ca2+ from increasing above its normal resting concentration. Thus an increase in intracellular Ca2+ is not necessary for zygote RVD; consistent with this, inhibitors of Ca(2+)-activated K+ channels had little or no effect on RVD. RVD in zygotes was also completely inhibited by millimolar amounts of extracellular ATP. ATP has been shown to inhibit current passed by the volume-sensitive organic osmolyte-Cl- channel in other cells, and thus zygotes may have such a channel participating in RVD.

Molecular characterization and differential mRNA tissue distribution of mouse apolipoprotein D.

The mouse apolipoprotein D gene was isolated from a brain cDNA library. The nucleotide sequence contains a unique reading frame coding for a protein sharing 79.5% homology with human apoD, 86.2% homology with rabbit apoD and 92.6% homology with rat apoD. The four sequences have two potential asparagine-linked glycosylation sites at residues 45 and 78, and possess the two consensus sequences of the lipocalin family which coincide with the most conserved regions in the four species studied. The distribution of apoD mRNA among mouse organs was determined by Northern blot and quantitative dot blot analysis. The highest levels of mRNA were found in the central nervous system (CNS), namely in the spinal cord, the cerebellum and the brain. Very low concentrations were detected in all the other organs tested. In some organs (spleen, kidney, intestines, heart), a second messenger of lower molecular weight was detected. Gene expression was also measured in rat tissues. As in the mouse, rat CNS was found to be by far the highest expressor of apoD mRNA, in contrast to the rabbit and human. Levels of expression in most mouse and rat organs appeared to be much lower than in the same organs of the rabbit and human. Since apoD is expressed at sites of nerve regeneration as well as apoE, our results raise the question of whether or not the two proteins play a coordinated role in the CNS.

The magnitude of acute and chronic alcohol abuse in trauma patients.

OBJECTIVE: To assess the incidence of acute alcohol intoxication and the proportion of trauma patients with evidence of chronic alcohol abuse. DESIGN: Prospective cohort study. SETTING: Regional level I trauma center. PARTICIPANTS: Patients aged 18 years and older admitted with blunt or penetrating trauma. MAIN OUTCOME MEASURES: Admission blood alcohol concentrations (BACs), the Short Michigan Alcohol Screening Test (SMAST), and biochemical markers for chronic alcohol abuse. RESULTS: Of the 2657 patients enrolled, 47.0% had a positive BAC and 35.8% were intoxicated (BAC > or = 100 mg/dL) on admission to the emergency department. Intoxicated patients were more likely to be 25 to 34 years old, male, and nonwhite; the highest proportion of intoxicated patients was among victims of stab wounds. Three fourths of acutely intoxicated patients had evidence of chronic alcoholism as indicated by a positive SMAST, and 25% to 35% of acutely intoxicated patients had biochemical evidence of chronic alcohol abuse. CONCLUSIONS: The high prevalence of both acute intoxication and chronic alcoholism in trauma patients indicates the need to diagnose and appropriately treat this pervasive problem in trauma victims.

A descriptive study of trauma, alcohol, and alcoholism in young adults.

Young adults, 18-20 years of age, admitted to a trauma center via the emergency department, were studied to determine if they had been drinking prior to their injury event. The prevalence of self-reported chronic alcohol problems was examined using the short Michigan Alcohol Screening Test (SMAST). Of the 319 subjects, 131 (41%) tested positive for alcohol, including about one-half of those with intentional injuries and 38% with unintentional injuries. Approximately 22% had blood alcohol concentrations of 100 mg/dL or more, indicating they were legally intoxicated at the time of their injury. Of study subjects who completed the SMAST, 49% attained scores suggesting potential or probable alcoholism, and 20% had already sought some type of treatment, despite their young age. Health-care practices and policies related to these findings include routine screening of trauma patients for alcohol abuse and integration of chemical dependency intervention services with trauma care.

Effects of alcohol intoxication on the initial assessment of trauma patients.

STUDY OBJECTIVES: To evaluate the influence of alcohol intoxication on the initial assessment and treatment of trauma patients. DESIGN: A prospective study of 2,237 trauma patients 18 years of age or older admitted to a Level I trauma center over a 19-month period. RESULTS: The study population was primarily male (78%) and white (73%) and had sustained blunt trauma (79%). One thousand fifty-three patients (47.1%) had positive blood alcohol concentration (BAC); median BAC in patients with any detectable alcohol was 179 mg/dL. When stratified by injury severity categories and compared with nonintoxicated (BAC less than 100 mg/dL) patients, intoxicated patients with an Injury Severity Score (ISS) of 1 to 15 were more likely to undergo the following: field and/or ED intubation (relative risk [RR], 2.22; 95% confidence interval [CI], 1.7 to 2.7); diagnostic peritoneal lavage (RR, 1.83; CI, 1.43 to 2.3); head computed tomography scanning (RR, 1.18; CI, 1.0 to 1.4); and intracranial pressure monitoring (RR, 1.41; CI, 0.74 to 2.7). The effects were less pronounced for those patients with an ISS of more than 15, except for intracranial pressure monitoring where patients with an ISS of more than 15 were 47% more likely to have intracranial pressure monitoring if intoxicated (RR, 1.47; CI, 1.2 to 1.9). CONCLUSION: Acute intoxication appears to alter the initial assessment of injury severity, resulting in an increased use of invasive diagnostic and therapeutic procedures.

[Therapeutic approach in chronic maxillary sinusitis. Apropos of 469 patients and 640 sinuses]. / Approche thérapeutique des sinusites maxillaires chroniques. A propos de 469 patients et de 640 sinus.

Between 1982 and 1987 we treated 469 patients who accounted for a total of 640 involved sinuses. These patients suffered from various stages of chronic sinusitis. Endoscopy of the maxillary sinuses enabled us to collect important information regarding the state of the mucosa and secretions, to collect bacteriological samples and to inspect the ostium and take pressure measurements. There was an 80% success rate in cases of sinusitis treated by drainage. In our opinion the indication for inferior meatotomy is as an extraction procedure and is not of value for aeration drainages. Nevertheless, out of a total of 141 inferior meatotomies performed, 80% remained patent after a mean follow up period of 3 years.

[Indications, technics and results of middle meatotomy. Apropos of 94 cases]. / Indications, techniques, résultats des méatotomies moyennes. A propos de 94 cas.

Middle meatotomies were essentially performed during the second period of our study from 1985 to 1987. The typical indication was for closed sinusitis, in order to allow aeration and satisfactory drainage. This was performed with an optical system at 30 degrees. The technique used was an anterior approach via total unciformectomy or a posterior approach using an Ostrum forceps. Good results, i.e. meatotomies which remained patent, were obtained in 80% of cases. The mean period of follow up was 3 years (6 months to 4 years). There were few complications overall for this form of endonasal surgery, though the possibility of damage to the lacrymal duct does exist. Synechiae were the most important functional complication and need to be prevented by meticulous post-operative care.

[Usefulness of the laser in post-radiotherapy arytenoid edema]. / Utilité du laser dans les oedèmes aryténoïdiens post-radiothérapiques.

Laser treatment of post-radiotherapy arytenoid edema was applied to two groups of patients with either laryngeal mobility or immobility. When mobility was normal the technic was simple, since only resection of the arytenoid edema was necessary and results were always excellent. In the second group, resection of arytenoid edema must be combined with partial or total resection of arytenoid, but in this case there is a risk of chondritis. Results in this series were excellent and only one case of post-laser radionecrosis was noted.

[Sinus endoscopy in the diagnosis of rhinosinal pathology]. / Apport de la sinusoscopie dans le diagnostic de la pathologie rhino-sinusienne.

Findings in 86 endoscopic examinations of the nose and paranasal sinuses were compared with clinical and radiographic signs. Precise evaluations of endomaxillary lesions, including analysis of secretions, and histologic examinations were conducted. Clinical and endoscopic findings failed to concord in 28 p. cent of cases, and radiological and endoscopic images in 26 p. cent of cases, the majority of these errors arising because of homogeneous total filling of the sinuses. In spite of these divergences, endoscopy is relatively non-invasive and simple to perform, and occupies a place of choice in the diagnosis of disorders of the nose and sinuses. Sinus endoscopy can also be of therapeutic value, particularly for aeration of the sinuses and during minor surgical procedures.

[Ewing's inverted nasosinal papilloma. Critical study of therapeutic indications. Apropos of 14 cases]. / Papillomes inverses naso-sinusiens d'Ewing. Etude critique des indications thérapeutiques. A propos de 14 cas.

Fourteen cases of Ewing's inversed papilloma of the nose and sinuses were treated as primary lesions or after recurrence. Clinical and paraclinical, mainly radiologic, endoscopic, and biologic investigations are of fundamental importance and should be as extensive for primary as for recurrent lesions. Treatment is largely dependent on the results of these examinations, is essentially surgical, and should respond to two imperatives. First, it should lead to total excision under the best possible conditions, and second, it should involve a minimum of sequelae, particular esthetic complications. Techniques for excision through the natural pathways are described, followed by those using an external approach (canine fossa or para-lateronasal approaches), as well as their advantages and inconveniences. The preferred procedure is through an open, wide, para-lateronasal approach, enabling the most effective excision to be performed, this technique appearing to be essential for recurrent lesions.

[Comparison of a series of tympanic homografts alone and moulded autografts]. / Comparaison d'une série d'homogreffes de tympan seul et d'autogreffes moulées.

Anatomical and functional results were compared in a series of 60 tympanic homografts alone with those obtained after 80 moulded autografts. Anatomically, the moulded autografts were definitely superior complications of microperforations in approximately one-third of cases. However, from the functional point of veiw, homografts appear to provide better results, particular when the ossicles are intact.