Re-emergence of brucellosis in cattle in France and risk for human health.

A case of human brucellosis was diagnosed in France in January 2012. The investigation demonstrated that the case had been contaminated by raw milk cheese from a neighbouring dairy farm. As France has been officially free of bovine brucellosis since 2005, veterinary investigations are being conducted to determine the origin of the infection and avoid its spread among other herds. Hypotheses about the source of this infection are discussed.

Optimal combination of estimating equations in the analysis of multilevel nested correlated data.

Multilevel nested, correlated data often arise in biomedical research. Examples include teeth nested within quadrants in a mouth or students nested within classrooms in schools. In some settings, cluster sizes may be large relative to the number of independent clusters and the degree of correlation may vary across clusters. When cluster sizes are large, fitting marginal regression models using Generalized Estimating Equations with flexible correlation structures that reflect the nested structure may fail to converge and result in unstable covariance estimates. Also, the use of patterned, nested working correlation structures may not be efficient when correlation varies across clusters. This paper describes a flexible marginal regression modeling approach based on an optimal combination of estimating equations. Particular within-cluster and between-cluster data contrasts are used without specification of the working covariance structure and without estimation of covariance parameters. The method involves estimation of the covariance matrix only for the vector of component estimating equations (which is typically of small dimension) rather than the covariance matrix of the observations within a cluster (which may be of large dimension). In settings where the number of clusters is large relative to the cluster size, the method is stable and is highly efficient, while maintaining appropriate coverage levels. Performance of the method is investigated with simulation studies and an application to a periodontal study.

A Prospective Cohort Study of Bisphenol A Exposure from Dental Treatment.

Laboratory studies show that bisphenol A (BPA) leaches from bisphenol A-glycidyl methacrylate (bisGMA)-based dental materials. We aimed to quantify the extent to which children are exposed to BPA from dental treatment with bisGMA materials, by amount of treatment and type of sedation. We hypothesized that posttreatment urinary BPA (uBPA) concentrations would be higher among patients with more surfaces treated with bisGMA-based materials and among patients receiving general anesthesia compared with pretreatment concentrations. We conducted a prospective cohort study in 211 children, 4 to 12 y old, who had no prior resin-based dental treatment. We measured uBPA concentrations twice before treatment and at 2 d and 1, 4, and 16 wk posttreatment. We abstracted treatment data (surfaces treated) from the chart. We generated descriptive statistics and compared pre- and posttreatment uBPA concentrations using generalized estimating equations. Participants were 51% female, 46% non-White, and 74% publicly insured. The median age was 6 y. The mean number of tooth surfaces exposed to BisGMA materials (composites/sealants) was 7.5 (SD 5.3). Overall, uBPA concentrations were 86% higher (95% confidence interval [CI] 42% to 143%, P < 0.001) at 2 d posttreatment compared with pretreatment concentrations. The uBPA concentrations 2 d posttreatment versus pretreatment tended to be higher (112%, 95% CI 53% to 194%) among those receiving treatment on >4 surfaces than those receiving treatment on ≤4 surfaces (50%, 95% CI -2% to 130%). Two days after treatment, uBPA was significantly higher than pretreatment concentrations in children receiving nitrous oxide but not in those receiving general anesthesia. Among all findings, uBPA concentrations returned to baseline by 4 wk. Children experience short-term increases in BPA from dental treatment. The impact of relatively high, short-term BPA exposure on child health is unknown. Given the widespread use of BisGMA-based dental materials and that chronic low-dose BPA exposure may adversely affect child health, strategies that minimize BPA exposure could potentially improve child health.

Invasive arterial blood pressure monitoring in an out-of-hospital setting: an observational study.

BACKGROUND: Non-invasive arterial blood pressure measurement is often inaccurate in emergency unstable patients. A study was undertaken to assess the feasibility of out-of-hospital intra-arterial catheterisation in haemodynamically unstable patients and to evaluate the correlation between invasive and non-invasive arterial pressure values. METHODS: In this prospective 2-year observational study conducted by mobile emergency medical units, the success rate of arterial catheterisation was calculated and blood pressure values measured invasively and non-invasively after successful catheterisation were compared. RESULTS: 94 patients were included. The success rate for catheterisation (44 radial access, 50 femoral access) was 86% (95% CI 79% to 93%). Bias and precision in invasive versus non-invasive comparisons were -0.1, 38 mm Hg for systolic pressure and 4.2, 27 mm Hg for diastolic pressure. Values differed by more than 20 mm Hg in over 40% of patients. Invasive measurement led to 79 changes in vasoactive treatment in 51 patients. CONCLUSION: Emergency out-of-hospital invasive arterial blood pressure monitoring in haemodynamically unstable patients is highly feasible. Discrepancies between invasive and non-invasive measurements are common and highlight the value of early out-of-hospital monitoring.

A study of optic nerve head drusen in 38 pseudoxanthoma elasticum (PXE) patients (64 eyes). Location of optic nerve head drusen in PXE.

PURPOSE: To investigate the prevalence and location of optic nerve head drusen and their potential association with other PXE-related ophthalmic abnormalities. MATERIALS AND METHODS: Thirty-eight of the 155 patients (57 male and 98 female aged 49±17 years) included in this retrospective study had optic nerve head drusen. All of the patients underwent a comprehensive ophthalmic examination, including color images using red-free, blue and red filters, autofluorescence imaging and late-phase ICG frames. Comparative analysis of both groups (optic nerve head drusen or not) was conducted using R statistical software. RESULTS: The prevalence of optic nerve head drusen in our cohort was 24.5%. In this study, no evidence of a significant link between optic nerve head drusen and other fundus abnormalities was detected. They were more commonly located in the nasal sector than in the temporal sector of the optic disc (P<0.001). They were more frequently situated superonasally than inferonasally (P<0.004), superotemporally (P<0.001) or inferotemporally (P<0.03). No central visual field defect was observed in OND+ patients who were unaffected by macular disorders. DISCUSSION: We hypothesized this predominantly nasal primary location may result from greater sensitivity in the nasal optic nerve fibers which follow a much more angular path once they arrive in the scleral canal, accounting for accumulation of axoplasmic debris. CONCLUSION: In PXE, optic nerve head drusen are mostly located in the superonasal quadrant, causing progressive optic nerve invasion but probably no central visual field defects.

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. There were no statistically significant differences between treatment groups in the numbers of bacteria growing on antibiotic- or mercury-supplemented plates. This study provided no evidence that amalgam fillings on posterior teeth influenced the level of antibiotic- or mercury-resistant oral or urinary bacteria as detected by culture.

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. RESULTS: A total of 25 questionnaires were obtained from teachers who participated in the programme. Ninety-two per cent of the respondents had a positive opinion concerning the existence of the programme. Eighty-eight per cent of the teachers believed that the programme changed student's knowledge about dental hygiene. Ninety-two per cent of teachers supported the existence of the programme and 88% of them disagreed with a statement that participation in the programme was a waste of time. Teachers who did not collaborate actively with dental hygiene activities indicated belief that the programme affected school activities (P = 0.003). Teachers who actively participated in the programme believe that dental hygiene activities were important for students (P = 0.005). CONCLUSIONS: Teacher evaluations of this kind of programme are critical for the development of school-based Dental Hygiene Education programmes. Teachers believe that Dental Hygiene Education is crucial for students' well-being.

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. Community activities involved participants and their fellow students and were aimed at providing education on oral health in a school environment. METHODS: Dental hygienists developed clinical work with prophylaxis, sealants application and topical fluoride and implemented the community programme with in-class sessions on oral health themes. Twice a month fluoride mouthrinses and bi-annual tooth brushing instructional activity took place. Participation at dental-hygiene activities, sealed teeth with no need of restoration and dental-plaque-index were measures used to evaluate success of the programme for the participants. RESULTS: Improvement in dental hygiene is shown by the decrease in dental plaque index scores (P < 0.0001); also sealants integrity is achieved in 86.3% of teeth. 888 (13.7%) teeth with sealants had to be restored or were lost. Children participated actively on dental hygiene activities. Teachers became aware of the problem and included oral-health in school curricula. CONCLUSION: Dental hygiene activities have shown to be helpful to promote dental hygiene, promote oral health and to provide school-age children with education on habits that will be important for their future good health.

[Systematic hearing screening for newborns in the Champagne-Ardennes region: 32,500 births in 2 years of experience]. / Dépistage auditif néonatal systématique en région Champagne-Ardenne: à propos de 30500 naissances en deux années d'expérience.

OBJECTIVES: To report a Universal Newborn Hearing Screening (UNHS) program developed in the Champagne-Ardennes region in 2004-2005. METHODS: A team of ENT specialists and pediatricians set up a UNHS program designed to reduce the age of diagnosis and care of bilateral congenital deafness. The program was mainly based on automated acoustic otoacoustic emissions and a strict follow-up by the Regional Neonatal Screening Center. RESULTS: In 2004 and 2005, 29,944 neonates from 30,518 births were screened (98.11%). Of the neonates screened, 409 (1.38%) failed the test and were referred. The average retest delay was 2 weeks. Eleven were lost to follow-up, 371 (94%) had a successful second test on one or both ears, 27 (7%) failed the test a second time and had a diagnosis of ABR. Twenty-four cases of bilateral deafness were identified early, 14 of which had no risk factors. One of the children lost to follow-up was actually deaf, which was diagnosed at 18 months of age. Since the beginning of the UNHS program, the average age of diagnosis was lowered to less than 3 months. CONCLUSION: Our experience tends to demonstrate that UNHS is possible and the program allows an early diagnosis of bilateral congenital hearing loss.

[Introduction of the intubating Laryngeal Mask Airway in a prehospital emergency medical unit]. / Mise en place du masque Laryngé-Fastrach au sein d'un service médical d'urgence et de réanimation.

OBJECTIVES: Assessment of the intubating Laryngeal Mask Airway(trade mark) in a prehospital emergency mobile unit (PEMU). STUDY DESIGN: Observational study. METHODS: All the emergency physicians and nurses of the PEMU were trained with the intubating laryngeal mask (ILMA) handling on manikin and a learning curve was carried out. One year after the initial formation, a reassessment of the operators was performed. Following the initial formation, all the vehicles of the PEMU were equipped with ILMA and during 15 months all cases of ILMA use were recorded. The success rate and the difficulties met were analysed. RESULTS: Initial formation on manikin showed that at least 8 handling of the device were mandatory to achieve a 100% success rate. A significant reduction of tracheal tube insertion delay was observed up to the eight manipulations. One year after the initial formation, a significant loss of performance was observed. Over the clinical study period 20 ILMA were used with adequate ventilation through the mask in all cases and a possible intubation in 80% of the patients. CONCLUSION: The ILMA is a potential useful device in the prehospital setting. Initial formation and maintenance of the skill acquired with this technique are essential.

Site-Specific Neutrophil Migration and CXCL2 Expression in Periodontal Tissue.

The oral microbial community is the best-characterized bacterial ecosystem in the human host. It has been shown in the mouse that oral commensal bacteria significantly contribute to clinically healthy periodontal homeostasis by influencing the number of neutrophils that migrate from the vasculature to the junctional epithelium. Furthermore, in clinically healthy tissue, the neutrophil response to oral commensal bacteria is associated with the select expression of the neutrophil chemokine CXCL2 but not CXCL1. This preliminary study examined the contribution of commensal bacteria on neutrophil location across the tooth/gingival interface. Tissue sections from the root associated mesial (anterior) of the second molar to the root associated distal (posterior) of the second molar were examined for neutrophils and the expression of the neutrophil chemokine ligands CXCL1 and CXCL2. It was found that both the number of neutrophils as well as the expression of CXCL2 but not CXCL1 was significantly increased in tissue sections close to the interdental region, consistent with the notion of select tissue expression patterns for neutrophil chemokine expression and subsequent neutrophil location. Furthermore, mice gavaged with either oral Streptococcus or Lactobacillus sp. bacteria induced a location pattern of neutrophils and CXCL2 expression similar to the normal oral flora. These data indicate for the first time select neutrophil location and chemokine expression patterns associated with clinically healthy tissue. The results reveal an increased inflammatory load upon approaching the interproximal region, which is consistent with the observation that the interproximal region often reveals early clinical signs of periodontal disease.

Circadian variation in urinary excretion of bone collagen cross-links.

Bone resorption can be evaluated by measuring the urinary excretion of collagen type I cross-linked N telopeptides (NTx). Since it is difficult to obtain (and verify) 24 h urine collections from patients, untimed spot urines are more practical. Such measurements, however, need correction for urine dilution and potentially may vary with collection time since a circadian rhythm in bone metabolism has been reported. This study examined cross-link excretion in urine voids serially collected during a 24 h period from subjects living their normal daily routine (as opposed to a controlled hospital setting). This mimics the situation for walk-in patients visiting a clinician and providing a spot urine. A total of 35 dentists (20 males, 15 females) collected all urine voids separately over a 24 h period. Urines were analyzed for creatinine and NTx. The effects of time of day on the excretion rates of these metabolites (in nmol/h) and on the cross-link:creatinine ratio were assessed. A circadian rhythm was evident in the excretion rate of creatinine with a peak in the late afternoon (18% higher than the 24 h mean, p = 0.0004). The NTx excretion rate peaked in the morning (9% higher than the 24 h mean) but this latter rhythm was not statistically significant (p = 0.31). The NTx:creatinine ratio fell during the day from a high (122% of the 24 h mean) in the early morning to a low in the early evening. This rhythm in the NTx:creatinine ratio in untimed spot urines was statistically significant (p < 0.0001). In conclusion, the NTx:creatinine ratio in spot urines from adult outpatient subjects showed a significant circadian rhythm. Variations in creatinine excretion were the primary cause. Time of day should, therefore, be taken into account when comparing test results of spot urines with normal ranges or with other samples from the same subject.

AMESFIT: a microcomputer program for fitting linear-exponential dose-response models in the Ames Salmonella assay.

The Ames Salmonella/microsome assay remains the most widely used microbial test for genotoxicity. In this article, we describe a microcomputer program developed to fit a linear-exponential dose-response model to Ames assay data for established mutagens. The model includes a linear term to describe the mutagenic effects of the test agent at low to moderate doses and an exponential attenuation factor to accommodate downturns at high doses due to cytotoxicity. Quasi-likelihood methods are used to obtain estimates of the unknown model parameters, thereby avoiding the need to fully specify the distribution of the experimental data. This method of estimation also allows for extra-Poisson variation that is characteristic of counts of mutant colonies of bacteria observed in the Ames assay. The particular linear-exponential model used here was developed for use in the analysis of a recent large-scale collaborative trial using the Ames assay sponsored by the International Programme on Chemical Safety. The use of our program is illustrated using sample data sets taken from that collaborative study.

Between-subject and within-subject statistical information in dental research.

The evaluation of risk factors in dental research frequently uses observations at multiple sites in the same patient. For this reason, statistical methods that accommodate correlated data are generally used to assess the significance of the risk factors (e.g., generalized estimating equations, generalized linear mixed models). In applications of these methods, it is typically assumed (implicitly, if not explicitly) that between-subject and within-subject comparisons will produce the same estimated effect of the risk factor. When between- and within-subject comparisons conflict, the statistical methods can give biased estimates or results that are difficult to interpret. For illustration, we present two examples from periodontal disease studies in which different statistical methods give different estimates and significance levels for a risk factor. Statistical analyses in dental research should assess whether different sources of information give similar conclusions about risk factors or treatments.

Within-subject coronal caries distribution patterns: an evaluation of randomness with respect to the midline.

The distribution of caries among homologous surfaces can exhibit three possible patterns: random, aggregated, or regular. In a random caries pattern, caries lesions are randomly distributed among homologous surfaces. An aggregated caries pattern is distinguished by the aggregation of lesions on one side of the mouth or the other to a greater extent than would be expected by chance alone. For a regular caries pattern, the left-right distribution of lesions is more homogenous than would be expected by chance alone. A test statistic based on the left-right distribution of caries lesions among discordant homologous pairs was developed to investigate which of these three caries patterns is present in a representative sample of the adult United States population. The data originated from the National Survey of Oral Health in the US (Employed Adults), 1985-1986. Of the 15,132 subjects studied, 12,776 subjects had 2 or more decayed or filled teeth. (At least 2 carious or filled teeth are required for detection of patterns.) Approximately 50% of these subjects (n = 6,439) had two or more discordant homologous tooth pairs. With these tooth pairs, the hypothesis of a random caries pattern was rejected in favor of an aggregated caries pattern (p < 0.0001). Similar findings were obtained with discordant homologous surface pairs. This aggregation of caries on one side of the mouth or the other may be due to genetic, infectious, and/or environmental factors.

Evaluating the validity of probing attachment loss as a surrogate for tooth mortality in a clinical trial on the elderly.

Most periodontal trials are based on the assumption that the superior treatment, as judged by short-term intangible changes in probing attachment levels (the surrogate), is also the treatment most likely to affect tooth mortality. This assumption is valid if: (1) the surrogate is informative about tooth mortality, and (2) the surrogate captures a substantial proportion of the treatment effect on tooth mortality (e.g., > 50% or 75%). The goal of this study was to evaluate whether both conditions were satisfied in a randomized controlled trial (RCT) of elders at high risk for dental diseases. The results suggested that the first condition for a valid surrogate was satisfied: Both one- and two-year changes in probing attachment level were informative about tooth mortality risk. A 1-mm loss measured over a one-year period was associated with a 56% increased tooth mortality risk (relative risk = 1.56; 95% confidence interval, 1.08 to 2.26; p = 0.017); a 1-mm loss measured over a two-year period was associated with a 102% increased risk for tooth mortality (relative risk = 2.02; 95% confidence interval, 1.26 to 3.25; p = 0.004). The second condition necessary for a valid surrogate could not be confirmed in the present trial. With 95% confidence, it was concluded that one-year changes in probing attachment level measurements did not capture a significant proportion of the treatment effect (point estimate, 6%; 95% confidence interval;-38% to 53%). No useful statements could be made regarding the proportion of treatment effect captured by two-year changes, due to the width of the confidence interval (point estimate, 18%; 95% confidence interval;-151% to 140%). It is concluded that (1) the evidence surrounding the one-year change in probing attachment level indicates that it can be ruled out as being anything more than a weak surrogate marker for tooth mortality, and (2) further research is required to study the validity of two-year change in probing attachment level as a surrogate marker. Due to characteristics of the population and the treatments investigated, the generalizability of these findings to other RCTs is questionable.

The estimation of caries prevalence in small areas.

National surveys have been effective for the estimation of caries prevalence in broad regions of the US. However, it is unclear if data from such surveys can be used to estimate prevalences in small areas such as states or counties because of small sample sizes within individual areas. In this study, we applied specialized statistical methods to the estimation of small-area caries measures using data from an oral health survey conducted in the State of Washington. Dental exams to assess caries and the presence of sealants and fluorosis were performed on 2921 third-grade students in 84 public schools selected by a stratified random sample from all 39 counties in the state. Statistical methods for small-area estimation were used to estimate disease and sealant utilization measures for each of the counties. Adjustment was made for covariates measured at the school level, including ethnicity and the proportion of children in the Federally sponsored school lunch program. Substantial variability in disease and sealant utilization between counties was found. The estimated number of decayed and filled surfaces per child was 4.7 (inter-county range, 2.4 to 7.4). The estimated number of surfaces of untreated decay was 1.2 per child overall (range, 0.5 to 3.1). Thirty percent of the children had restorative treatment needs (range, 15 to 54%). The prevalence of sealants on one or more permanent molars was estimated to be 34% (range, 19 to 46%). Overall, only 8% of children showed evidence of fluorosis. The results demonstrate the usefulness of small-area estimation methods for oral health surveys.

Forty-five-year tooth survival probabilities among men in Oslo, Norway.

Changes in tooth survival probabilities over a person's lifetime have remained largely unexplored. The goal of this study was to evaluate changes in the 45-year tooth survival probabilities in a cohort of 565 Norwegian males who were examined in 1969 as young adults, and followed up into mid-life (examination years and sample sizes (n): 1971 (n = 381), 1973 (n = 292), 1975 (n = 245), 1981 (n = 228), 1988 (n = 202), and 1995 (n = 223). The results indicated that the tooth survival probabilities varied considerably both (i) among teeth within individuals, and (ii) over time. The 45-year survival probabilities for the 28 teeth fell into the following ranges: larger than 95% for incisors and cuspids; between 84% and 92% for premolars; and between 59% and 96% for molars. Over the first 4 post-eruptive decades, the tooth mortality risks (excluding orthodontic extractions) were: 1st decade, 2.0% (from 1.7 to 2.4%); 2nd decade, 0.2% (from 0.1 to 0.4%); 3rd decade, 0.6% (from 0.4 to 0.8%); and 4th decade, 1.1% (from 0.8 to 1.5%). The tooth mortality risks in the 2nd, 3rd, and 4th decades were probably somewhat underestimated (due to dropout bias), suggesting that the true underlying tooth mortality hazard function may have been V-shaped. The conclusions were that the tooth mortality hazard during the first 4 post-eruptive decades was bathtub-shaped and that it varied considerably among teeth within individuals.

Laparoscopic splenectomy in patients with hematologic malignancies.

BACKGROUND: Although laparoscopic splenectomy (LS) for benign hematologic disease is well accepted, its role in hematologic malignancies is not clearly defined. This study examined the efficacy and feasibility of LS for hematologic malignancies. METHODS: Records were reviewed from patients who underwent LS at two university hospitals. Charts from 77 open splenectomies for malignancy (OM) during the same period were also reviewed. RESULTS: Fifty-three patients underwent LS, 22 for hematologic malignancies (LM) and 31 for benign hematologic disorders (LB). Median splenic weight was greater in the LM group (930 g) than in the LB group (164 g, P = 0.001). LM was associated with longer operations and greater blood loss than was LB. LM had a 41% conversion rate. Morbidity, mortality, and transfusion rates were similar. Median hospital stay was shorter for LM (4 days) than for OM (6 days, P = 0.001). CONCLUSIONS: LS is feasible in hematologic malignancies but is associated with increased operative time and blood loss and a high conversion rate. Morbidity and mortality, however, was similar. Shorter hospital stays for LM compared with OM may translate into earlier recovery and initiation of antineoplastic therapy.

Sources of variation in the mutagenic potency of complex chemical mixtures based on the Salmonella/microsome assay.

Twenty laboratories worldwide participated in a collaborative trial sponsored by the International Programme on Chemical Safety on the mutagenicity of complex mixtures as expressed in the Salmonella/microsome assay. The U.S. National Institute of Standards and Technology provided homogeneous reference samples of urban air and diesel particles and a coal tar solution to each participating laboratory, along with samples of benzo[a]pyrene and 1-nitropyrene which served as positive controls. Mutagenic potency was characterized by the slope of the initial linear component of the dose-response curve. Analysis of variance revealed significant interlaboratory variation in mutagenic potency, which accounted for 57-96% of the total variance on a logarithmic scale, depending on the sample, strain and activation conditions. Variation among replicate extractions of organic material (required for the air and diesel particles) and among replicate bioassays within the same laboratory was also appreciable. The average potencies for air and diesel particles in laboratories using Soxhlet extracts were not significantly different from those in laboratories using sonication, although there was larger interlaboratory variation for the Soxhlet method. Repeatability (which approximates the coefficient of variation within laboratories) ranged from 18 to 40% for air and diesel particles extracted using sonication, depending on the strain and activation conditions. Repeatability of Soxhlet-extracted air and diesel particles, however, ranged from about 37 to 89% including outliers and from about 11 to 31% excluding outliers. Repeatability of the coal tar sample and the 2 positive controls was in the range 18-34%. Reproducibility (which approximates the coefficient of variation between laboratories) was generally at least twice repeatability, and exceeded 100% for Soxhlet-extracted air and diesel particles, as well as 1-nitropyrene. Reanalysis of the data omitting observations of more than 1500 revertants/plate generally had little effect on these results. Elimination of outlying observations had limited impact, with the exception of Soxhlet-extracted air and diesel particles. In this case, reproducibility of bioassay results was notably improved, due largely to the omission of results for replicate extractions which varied more than 5-fold within one laboratory. Normalization of the log potency slopes for the mixtures by the corresponding slopes for benzo[a]pyrene tended to reduce this variation, although variation was increased after normalization by 1-nitropyrene. Adjustment for the percentage of organic matter extracted from the air and diesel particulate samples had little effect on variation for sonication-extracted particles, whereas variation was reduced for diesel particles and increased for air particles for Soxhlet.