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Background: Here we compare the performance of the high-throughput BD COR System (COR) to the Viper LT System (Viper) using the BD Onclarity HPV assay. Research Design and Methods: Remnant clinical specimens, contrived specimens in SurePath (BD) and PreservCyt (Hologic) media, and prospective clinical specimens in BD Cervical Brush Diluent (CBD) were tested. Outcomes included intra-laboratory agreement of Onclarity results on COR and inter-system agreement between COR and Viper. Results: Onclarity reproducibility on COR resulted in standard deviation and correlation of variation of Ct values ranging from 0.14 to 1.98 and 0.49% to 2.15%, respectively, for contrived specimens, and 0.9–3.08 and 2.89–9.21%, respectively, for clinical specimens. In the COR and Viper clinical agreement study, OPA for Onclarity ranged from 97.1%-98.9%, depending on the collection media type. PPA values for pooled, HPV(+) specimens at low positive (C95), and moderate positive (3XC95) target concentrations were ≥95.0% and 100%, respectively; PPA values associated with HPV 16, 18, 31, 45, 33/58, 52, 35/39/68, 51, and 56/59/66, individually, ranged from 93.8%-100%. Conclusions: Onclarity performance on COR is equivalent to Viper, and is accurate and reproducible for detection of all high-risk HPV genotypes, with a throughput of 330 results from a single 8-hour shift.

Detection of Bordetella pertussis and Bordetella parapertussis using molecular methods is sensitive and specific with a short turnaround time compared to other diagnostic methods. In this multicenter study, we compared the performance of the Simplexa Bordetella Direct kit to those of other molecular assays in detecting and differentiating B. pertussis and B. parapertussis in nasopharyngeal swab specimens. The limits of detection (LODs) were 150 CFU/ml or 3 fg/μl of DNA for B. pertussis and 1,500 CFU/ml or 10 fg/μl of DNA for B. parapertussis. A total of 1,103 fresh and residual frozen specimens from eight clinical sites were tested. Combining the data from individual clinical sites using different comparative assays, the overall positive percent agreement (PPA) and negative percent agreement (NPA) for B. pertussis were 98.7% and 97.3%, respectively. The overall PPA and NPA for B. parapertussis were 96.7% and 100%, respectively. For prospective fresh specimens, the overall PPA and NPA for both targets were 97.7% and 99.3%, respectively. For retrospective frozen specimens, the overall PPA and NPA for both targets were 92.6% and 93.2%, respectively. The percentage of invalid results was 1.0%. A cross-reactivity study using 74 non-Bordetella bacterial species and five yeast species revealed that the Simplexa Bordetella Direct kit was 100% specific. The hands-on time and assay run time of the Simplexa Bordetella Direct kit are favorable compared to those of other commercial and laboratory-developed tests. In summary, the Simplexa Bordetella Direct kit has a performance comparable to those of other molecular assays for the detection of B. pertussis and B. parapertussis.

A 6-h and a 24-h preenrichment procedure were compared for their ability to recover Salmonella spp. from selected low-moisture dairy foods. The foods were artificially inoculated several days before analysis, and 20 replicate test portions per procedure from each food were examined in each experiment. Samples examined by the 6-h abbreviated procedure were preenriched for 6 h at 35°C in an air incubator or water bath and centrifuged at 4,100 × g for 10 min. Pellets were suspended in tetrathionate broth and incubated for 24 h at 35°C. For the 24-h standard procedure, test portions were preenriched for 24 h at 35°C in an air incubator, subcultured to tetrathionate broth, and incubated for 24 h at 35°C. Selective enrichment broths from both procedures were streaked onto selective agar plates, and presumptive Salmonella isolates were identified by conventional biochemical and serological tests. Recovery of Salmonella spp. from instant nonfat dry milk and dry whole milk was equivalent for both preenrichment procedures. However, the relative effectiveness of the two procedures varied in the recovery of Salmonella spp. from noninstant nonfat dry milk, lactic casein, and rennet casein.

Two enzyme immunoassays (Salmonella-Tek™ and Report™) were compared with the standard culture method of the Association of Official Analytical Chemists (AOAC) and the Food and Drug Administration's Bacteriological Analytical Manual (BAM) for the recovery of Salmonella spp. from four low-moisture foods. Two protocols were used to compare the effectiveness of the two immunoassays: i) foods were contaminated in the dry state; or ii) serial tenfold dilutions of Salmonella spp. were inoculated into the postenrichments after incubation. Of three hundred 25-g test portions inoculated in the dry state, 199 gave confirmed positive reactions with the Salmonella-Tek™ assay, 193 with the Report™ assay, and 206 with the AOAC/BAM method. There were seven false-negative reactions with Salmonella-Tek™ and 13 false negatives with the Report™, a false negative being defined as one that was negative by the enzyme immunoassay but was confirmed positive by the AOAC/BAM culture method. When the postenrichments were inoculated after incubation, a lower number of cells gave a positive assay result with the Salmonella-Tek™ system than with the Report™ system, indicating greater sensitivity.

Five high-moisture foods were used to evaluate both the effect of a 6 h, rather than the standard 24 h, selective enrichment incubation period, and the efficiency of Rappaport-Vassiliadis (RV) medium relative to the use of selenite cystine (SC) and tetrathionate (TT) broths for the recovery of Salmonella . Cheese and lettuce were artificially inoculated with a pool of two serotypes, whereas the other foods were naturally contaminated. Significantly higher numbers of Salmonella -positive test portions were obtained at 24 h with the following food and media combinations: cheese (TT and RV media), lettuce (SC, TT, and RV media), raw chicken (RV medium), and pork sausage (SC, TT, and RV media). There were no significant differences between the two incubation periods in recovery of Salmonella from turkey. Overall, more Salmonella -positive test portions were obtained from samples of lettuce, chicken, and pork sausage selectively enriched in RV medium than in SC or TT broths. The results of this study indicate that not all high-moisture foods can be selectively enriched for 6 h without a significant loss in recovery of Salmonella . RV medium was superior to SC and TT broths for recovery of Salmonella from some meats and was at least as productive in its recovery from the other high-moisture foods tested.

Recovery of Salmonella from instant nonfat dry milk was studied by determining the efficiency of 6 and 24 h selective enrichment incubation periods, the productivity of Rappaport-Vassiliadis (RV) medium relative to selenite cystine and tetrathionate broths, and the potential enhancement of post enrichment with GN and M broths. Efficiency of the immunodiffusion method recently approved by the Association of Official Analytical Chemists (AOAC) for recovering Salmonella was also determined. Samples of artificially contaminated instant nonfat dry milk were selectively enriched for 6 h without compromising recovery. There was no advantage in using RV medium, and post enrichment did not enhance recovery. For the evaluation of the immunodiffusion method, three analysts independently read the test kit reactions. Of the 220 samples examined, 42.7-45.5% were positive by the immunodiffusion and the AOAC/Bacteriological Analytical Manual (BAM) culture methods, 42.3-43.2% were negative by both methods, 11.4-14.1% were negative by the immunodiffusion test and positive by the AOAC/BAM culture method, and 0-0.9% were negative by the AOAC/BAM method and positive by the immunodiffusion method.

Methodology was developed for isolation of Salmonella from skim milk, 2% fat milk, whole milk and buttermilk. Lactose broth, lactose broth plus brilliant green dye, buffered peptone water and each milk type plus brilliant green dye were evaluated as preenrichment broths. Incubation temperatures of 35 and 43°C were compared for use at the preenrichment stage. The recovery of Salmonella was determined after selective enrichment in selenite cystine, tetrathionate and Rappaport-Vassiliadis broths. Results indicated that fluid milk should be examined for Salmonella by being preenriched in lactose broth, subcultured to selenite cystine and tetrathionate broths and streaked to selective agars, with 35°C as the incubation temperature throughout the analysis.

Cultures of three Listeria monocytogenes serotypes and three Salmonella spp. were applied to the exterior surfaces of waxed cardboard or plastic milk containers. Contamination sites were sampled with premoistened cotton swabs during 14 d of refrigeration. Unstressed cells of Listeria survived up to 14 d on the surfaces of waxed (1 serotype) and plastic (3 serotypes) containers. Heat-stressed cells of all three serotypes of Listeria survived for 2 d on both types of containers. One serotype survived for 4 d, but only on plastic containers. Unstressed cells of all three Salmonella strains survived up to 14 d on both types of containers. Heat-stressed Salmonella strains survived up to 2 d (waxed containers) and 4 d (plastic containers).

Recovery of Salmonella species from dry whole milk, lactic casein, non-instantized nonfat dry milk, rennet casein and sodium caseinate was compared under rapid and slow conditions of rehydration. For rapid rehydration, a 25-g portion of each product was blended or swirled with 225 ml of appropriate preenrichment medium. After 60 min, the flask contents were adjusted to pH 6.8 and incubated at 35°C. For slow rehydration, a 25-g portion of each product was gently added to 225 ml of appropriate preenrichment medium, allowed to soak undisturbed for 60 min at room temperature, and then incubated at 35°C without pH adjustment. Recovery of Salmonella by the slow rehydration (soak) method was equal or enhanced for all products tested except sodium caseinate. Use of a meter instead of test paper to adjust the pH of rapid rehydration (blend/swirl) preenrichments did not improve recovery of Salmonella . Examination of dry whole milk and non-instantized nonfat dry milk by the soak method should be limited to 25-g amounts since 100-g and 375-g composites were not completely wetted. Composites of lactic and rennet casein weighing ⩽375 g, however, may be examined by the soak method without loss of analytical sensitivity.

Because damaged cells may account for a substantial proportion of the bacterial population in processed foods, the food microbiologist must choose the most appropriate methods for detecting damaged as well as noninjured cells. Any method intended to recover damaged organisms should include a resuscitative, or repair, process that will restore the injured cells to a sound physiological condition before subjecting them to the severity of selective enrichment media. It should also provide a reliable indication of the microbiological safety and quality of any particular food. This paper reviews various factors that affect the recovery of Salmonella spp., which include: (a) sample rehydration, (b) period of preenrichment, (c) incubation in both aerobic and anaerobic environments, (d) media composition and (e) the relative merits of preenrichment and direct selective enrichment. Because resuscitation of injured Salmonella cells does not occur during the selective enrichment step and beyond, the effect and interaction of these factors are considered primarily for the preenrichment step of the isolation procedure for Salmonella . This paper also reviews five methods recently developed for recovery of coliforms, which include: (a) hydrophobic grid membrane filtration, (b) radiometry, (c) electrical impedance, (d) fluorogenic assay and (e) the Petrifilm system. Each of these methods may incorporate a step for resuscitation of injured organisms.

International audience; 4,15-Diacetoxyscirpenol (DAS) is a mycotoxin primarily produced by Fusarium fungi and occurring predominantly in cereal grains. As requested by the European Commission, the EFSA Panel on Contaminants in the Food Chain (CONTAM) assessed the risk of DAS to human and animal health related to its presence in food and feed. Very limited information was available on toxicity and on toxicokinetics in experimental and farm animals. Due to the limitations in the available data set, human acute and chronic health-based guidance values (HBGV) were established based on data obtained in clinical trials of DAS as an anticancer agent (anguidine) after intravenous administration to cancer patients. The CONTAM Panel considered these data as informative for the hazard characterisation of DAS after oral exposure. The main adverse effects after acute and repeated exposure were emesis, with a no-observed-adverse-effect level (NOAEL) of 32 lg DAS/kg body weight (bw), and haematotoxicity, with a NOAEL of 65 lg DAS/kg bw, respectively. An acute reference dose (ARfD) of 3.2 lg DAS/kg bw and a tolerable daily intake (TDI) of 0.65 lg DAS/kg bw were established. Based on over 15,000 occurrence data, the highest acute and chronic dietary exposures were estimated to be 0.8 and 0.49 lg DAS/kg bw per day, respectively, and were not of health concern for humans. The limited information for poultry, pigs and dogs indicated a low risk for these animals at the estimated DAS exposure levels under current feeding practices, with the possible exception of fattening chicken. Assuming similar or lower sensitivity than for poultry, the risk was considered overall low for other farm and companion animal species for which no toxicity data were available. In consideration of the similarities of several trichothecenes and the likelihood of co-exposure via food and feed, it could be appropriate to perform a cumulative risk assessment for this group of substances.

International audience; Moniliformin (MON) is a mycotoxin with low molecular weight primarily produced by Fusarium fungi and occurring predominantly in cereal grains. Following a request of the European Commission, the CONTAM Panel assessed the risk of MON to human and animal health related to its presence in food and feed. The limited information available on toxicity and on toxicokinetics in experimental and farm animals indicated haematotoxicity and cardiotoxicity as major adverse health effects of MON. MON causes chromosome aberrations in vitro but no in vivo genotoxicity data and no carcinogenicity data were identified. Due to the limitations in the available toxicity data, human acute or chronic health-based guidance values (HBGV) could not be established. The margin of exposure (MOE) between the no-observed-adverse-effect level (NOAEL) of 6.0 mg/kg body weight (bw) for cardiotoxicity from a subacute study in rats and the acute upper bound (UB) dietary exposure estimates ranged between 4,000 and 73,000. The MOE between the lowest benchmark dose lower confidence limit (for a 5% response-BMDL 05) of 0.20 mg MON/kg bw per day for haematological hazards from a 28-day study in pigs and the chronic dietary human exposure estimates ranged between 370 and 5,000,000 for chronic dietary exposures. These MOEs indicate a low risk for human health but were associated with high uncertainty. The toxicity data available for poultry, pigs, and mink indicated a low or even negligible risk for these animals from exposure to MON in feed at the estimated exposure levels under current feeding practices. Assuming similar or lower sensitivity as for pigs, the CONTAM Panel considered a low or even negligible risk for the other animal species for which no toxicity data suitable for hazard characterisation were identified. Additional toxicity studies are needed and depending on their outcome, the collection of more occurrence data on MON in food and feed is recommended to enable a comprehensive human risk assessment.

The performance of the new ePlex Respiratory Pathogen (RP) panel (GenMark Diagnostics) for the simultaneous detection of 19 viruses (influenza A virus; influenza A H1 virus; influenza A 2009 H1 virus; influenza A H3 virus; influenza B virus; adenovirus; coronaviruses [HKU1, OC43, NL63, and 229E]; human rhinovirus/enterovirus; human metapneumovirus; parainfluenza viruses 1, 2, 3, and 4; and respiratory syncytial virus [RSV] [RSV subtype A and RSV subtype B]) and 2 bacteria ( Mycoplasma pneumoniae and Chlamydia pneumoniae ) was evaluated. Prospectively and retrospectively collected nasopharyngeal swab (NPS) specimens ( n = 2,908) were evaluated by using the ePlex RP panel, with the bioMérieux/BioFire FilmArray Respiratory Panel (BioFire RP) as the comparator method. Discordance analysis was performed by using target-specific PCRs and bidirectional sequencing. The reproducibility of the assay was evaluated by using reproducibility panels comprised of 6 pathogens. The overall agreement between the ePlex RP and BioFire RP results was >95% for all targets. Positive percent agreement with the BioFire RP result for viruses ranged from 85.1% (95% confidence interval [CI], 80.2% to 88.9%) to 95.1% (95% CI, 89.0% to 97.9%), while negative percent agreement values ranged from 99.5% (95% CI, 99.1% to 99.7%) to 99.8% (95% CI, 99.5% to 99.9%). Additional testing of discordant targets (12%; 349/2,908) confirmed the results of ePlex RP for 38% (131/349) of samples tested. Reproducibility was 100% for all targets tested, with the exception of adenovirus, for which reproducibilities were 91.6% at low virus concentrations and 100% at moderate virus concentrations. The ePlex RP panel offers a new, rapid, and sensitive “sample-to-answer” multiplex panel for the detection of the most common viral and bacterial respiratory pathogens.

Blended learning (BL) is the integration of classroom learning with an online environment. The purpose of this study was to determine whether dental students who experienced BL in a preclinical endodontic course demonstrated better manual skills, conceptual knowledge, and learning experience compared to those experiencing traditional learning. All eighty-one students (100 percent) in a preclinical endodontics course agreed to participate and were assigned to either the traditional or BL group. A root canal procedure was used to determine the level of manual skills gained by each group. Pre- and post-intervention quizzes were given to all students to evaluate conceptual knowledge gained, and the students' perspectives on the methods were evaluated with a survey. The BL group scored better than the traditional group on the manual skills exercise at a statistically significant level (p=0.0067). There were no differences in the post-intervention quiz scores between the two groups, and the students' opinions were positive regarding BL. With BL, the students were able to learn and demonstrate dental skills at a high level.

The histone chaperone Chromatin Assembly Factor 1 (CAF-1) deposits tetrameric (H3/H4)2 histones onto newly-synthesized DNA during DNA replication. To understand the mechanism of the tri-subunit CAF-1 complex in this process, we investigated the protein-protein interactions within the CAF-1-H3/H4 architecture using biophysical and biochemical approaches. Hydrogen/deuterium exchange and chemical cross-linking coupled to mass spectrometry reveal interactions that are essential for CAF-1 function in budding yeast, and importantly indicate that the Cac1 subunit functions as a scaffold within the CAF-1-H3/H4 complex. Cac1 alone not only binds H3/H4 with high affinity, but also promotes histone tetramerization independent of the other subunits. Moreover, we identify a minimal region in the C-terminus of Cac1, including the structured winged helix domain and glutamate/aspartate-rich domain, which is sufficient to induce (H3/H4)2 tetramerization. These findings reveal a key role of Cac1 in histone tetramerization, providing a new model for CAF-1-H3/H4 architecture and function during eukaryotic replication. DOI: http://dx.doi.org/10.7554/eLife.18023.001, eLife digest The DNA of a human, yeast or other eukaryotic cell is bound to proteins called histones to form repeating units called nucleosomes. Every time a eukaryotic cell divides, it must duplicate its DNA. Old histones are first removed from the nucleosomes before being re-assembled onto the newly duplicated DNA along with new histone proteins, producing a full complement of nucleosomes. A group of proteins called the chromatin assembly factor 1 (or CAF-1 for short) helps to assemble the histones onto the DNA. CAF-1 is made up of three proteins, and binds to two copies of each of the histones known as H3 and H4. These are the first histones to be assembled onto the nucleosomes. It was not clear how the components of CAF-1 are organized, or how CAF-1 recognizes histones. Liu et al. have now investigated the structure of CAF-1 and its interactions with the H3 and H4 histones by studying yeast proteins and cells. Yeast is a good model system because yeast CAF-1 is smaller and easier to isolate than human CAF-1, yet still performs the same essential activities. Using a combination of biochemical and biophysical techniques, Liu et al. found that one of the three proteins that makes up yeast CAF-1 – called Cac1 – forms a scaffold that supports the other CAF-1 proteins and histones H3 and H4. Moreover, a specific part of Cac1 is able to bind to these histones and assemble two copies of each of them to prepare for efficient nucleosome assembly. Further experiments revealed the specific areas where the CAF-1 proteins interact with each other and with the histones, determined how strong those interactions are, and confirmed that these interactions play important roles in yeast. Overall, the results presented by Liu et al. provide new insights into the structure of CAF-1 bound to H3 and H4. In order to understand in detail how CAF-1 helps to assemble histones onto DNA, future work needs to capture three-dimensional snapshots of the different steps in this process. Further investigation is also needed to discover how CAF-1 cooperates with other factors that promote DNA duplication. DOI: http://dx.doi.org/10.7554/eLife.18023.002

The microbiology laboratory offers a variety of procedures and diagnostic technologies to assist in the diagnosis of ocular infection. As many organisms cause eye infection, it is important to select the appropriate specimen for a particular ocular infection. Laboratories that receive specimens inoculated to primary culture media at the bedside or in the clinic will want to coordinate with those clinicians to provide collection and transport protocols along with fresh media. The types of specimens that may be taken for ocular infection include conjunctival swabs, corneal scrapings, aqueous and vitreous fluid aspirates, aspirates from an abscess or wound, periocular tissue, corneal donor ring in storage media, and contact lens solutions. Ocular pathogens include bacteria, fungi, acid-fast bacilli (AFB), viruses and parasites. Microbiology laboratory procedures that support the identification of ocular pathogens include smear and microscopy, conventional culture, serological assays, and a number of specific molecular assays to include nucleic acid amplification and DNA sequencing. Coordination between the clinician and the microbiologist is essential for proper clinical interpretation of results and resulting therapeutic management of the patient. Antimicrobial susceptibility testing (AST) yielding minimum inhibitory concentration (MIC) values with breakpoint interpretations is useful for therapeutic decision-making on patients requiring systemic therapy. MIC breakpoint interpretations are not defined for drugs that are applied topically or that are injected into the anterior or posterior segments of the eye, as concentrations achievable through direct administration can be significantly greater than those measured in serum after oral or parenteral administration.

Sharing resources through distance education has been proposed as one way to deal with a lack of full-time faculty members and maintain high-quality content in orthodontic residency programs. To keep distance education for orthodontic residents cost-effective while retaining interaction, a blended approach was developed that combines observation of web-based seminars with live post-seminar discussions. To evaluate this approach, a grant from the American Association of Orthodontists (AAO) opened free access during the 2009–10 academic year to twenty-five recorded seminars in four instructional sequences to all sixty-three orthodontic programs in the United States and Canada. The only requirement was to also participate in the evaluation. Just over half (52 percent) of the U.S. programs chose to participate; the primary reason for participating was because faculty members wanted their residents to have exposure to other faculty members and ideas. The non-participating programs cited technical and logistical problems and their own ability to teach these subjects satisfactorily as reasons. Although participating distant faculty members and residents were generally pleased with the experience, problems in both educational and technical aspects were observed. Educationally, the biggest problem was lack of distant resident preparation and expectation of a lecture rather than a seminar. Technically, the logistics of scheduling distant seminars and uneven quality of the audio and video recordings were the major concerns of both residents and faculty members. Proposed solutions to these educational and technical problems are discussed.

The purpose of our study was to evaluate the effectiveness of self-tests as a component of web-based self-instruction in predoctoral orthodontics and pediatric dentistry. To this end, the usage patterns of online teaching modules and self-tests by students enrolled in three courses at the University of North Carolina at Chapel Hill School of Dentistry were monitored and correlated to final exam grade and course average. We recorded the frequency of access to thirty relevant teaching modules and twenty-nine relevant self-tests for 157 second- and third-year D.D.S. students during the course of our data collection. There was a statistically significant positive correlation between frequency of accessing self-tests and course performance in one course that was totally based on self-instruction with seminars and multiple-choice examination (Level IV): Spearman correlation between frequency of self-test access and final exam grade, rho=0.23, p=0.044; correlation between frequency of self-test access and course average: rho=0.39, p=0.0004. In the other two courses we monitored, which included content beyond self-instruction with self-tests, the correlations were positive but not statistically significant. The students' use of online learning resources varied significantly from one course (Level I) to the next (Level II): Wilcoxon matched pairs signed-rank tests, S=-515.5, p=.0057 and S=1086, p

Kidney transplant from donors with hepatitis C virus (HCV) antibody has been limited to HCV viremic recipients only, due to concern of the HCV transmission. However, the new antiviral medications provide an opportunity to expand the utilization of these donors. To study the risk of HCV transmission in kidney transplantation, we used discarded donor kidneys and determined HCV RNA levels by quantitative real-time PCR in bilateral (right and left) kidney biopsies and plasma from 14 HCV antibody-positive donors (sensitivity: 15 international unit (IU)/mL plasma; 1.8 IU/50 nL kidney). In three NAT-negative donors, HCV RNA was negative in plasma and kidney. In all 11 NAT-positive donors, HCV RNA was positive in plasma (range: 5807-19 134 177 IU/mL) but negative in six kidneys from four donors with plasma HCV RNA

Five pre-enrichment methods were evaluated for effectiveness with the U.S. Food and Drug Administration's Bacteriological Analytical Manual Salmonella culture method in recovering S. Stanley, S. Poona, and S. Muenchen from artificially contaminated alfalfa seeds, and S. Saintpaul, S. Anatum, and S. Infantis from artificially contaminated mung bean seeds. The methods included: (1) Soak.Test portions were inoculated into pre-enrichment media; (2) Rinse.Test portions were rinsed with pre-enrichment media, and the media was decanted from the test portions; (3) Rinsed seed.Pre-enrichment media was added to the test portions that were rinsed in the rinse method; (4) Wet blend.Test portions were blended with the pre-enrichment media; and (5) Dry blend.Test portions were blended prior to pre-enrichment. The methods of pre-enrichment were also evaluated for effectiveness in recovering Pantoea agglomerans from alfalfa and mung bean seeds with a modified culture method for the recovery of Enterobacteriaceae from foods. The purpose of these studies was to provide a model for the recovery of Salmonella that may occur in seeds as a natural contaminant. The relative effectiveness of the soak method was consistently superior to the rinse method in isolating the selected Salmonella serovars from both seed types. Statistically, the rinsed seed method was as effective as the soak method in all trials, except 1 of 3, with S. Muenchen and alfalfa seeds (P > 0.05). The relative effectiveness of the methods in isolating P. agglomerans from alfalfa and mung bean seeds was similar to that observed with the artificially contaminated test portions. The soak method was consistently the most effective method and the rinse method was consistently the most ineffective method. The rinsed seed, wet blend, and dry blend methods were also as effective as the soak method in all 3 trials with each seed type (P > 0.05).

Increased understanding of the transcriptomic patterns underlying head and neck squamous cell carcinoma (HNSCC) can facilitate earlier diagnosis and better treatment outcomes. Integrating knowledge from multiple studies is necessary to identify fundamental, consistent gene expression signatures that distinguish HNSCC patient samples from disease-free samples, and particularly for detecting HNSCC at an early pathological stage. This study utilizes feature integration and heterogeneous ensemble modeling techniques to develop robust models for predicting HNSCC disease status in both microarray and RNA-seq datasets. Several alternative models demonstrated good performance, with MCC and AUC values exceeding 0.8. These models were also applied to discriminate between early pathological stage HNSCC and normal RNA-seq samples, showing encouraging results. The predictive modeling workflow was integrated into a software tool with a graphical user interface. This tool enables HNSCC researchers to harness frequently observed transcriptomic features and ensembles of previously developed models when investigating new HNSCC gene expression datasets.

The AmpliVue HSV 1+2 assay was compared to the ELVIS HSV ID and D 3 Typing Culture System for the qualitative detection and differentiation of herpes simplex virus 1 (HSV-1) and HSV-2 DNA in 1,351 cutaneous and mucocutaneous specimens. Compared to ELVIS, AmpliVue had sensitivities of 95.7 and 97.6% for detecting HSV-1 and HSV-2, respectively. Following arbitration of discordant results by an independent molecular method, the AmpliVue assay had a resolved sensitivity and specificity of 99.2 and 99.7%, respectively, for both HSV-1 and HSV-2, whereas ELVIS had a resolved sensitivity of 87.1% for HSV-1 and 84.5% for HSV-2.

Studies were conducted to determine the relative effectiveness of buffered peptone water (BPW), lactose (LAC) broth, and Universal Preenrichment (UP) broth for the recovery of Salmonella organisms from fruit rinses, whole fruit, and comminuted fruit. In the first phase, the relative effectiveness of the rinse and soak methods for the recovery of Salmonella from surface-contaminated mangoes and tomatoes was examined. Fruits were spot inoculated with single Salmonella serovars and held for 4 days at 26C before analysis was initiated. The contaminated fruit was rinsed in portions of BPW, LAC broth, or UP broth. Portions from each rinse were added to its respective broth (e.g., BPW to BPW). Individual whole fruit, in their remaining broth rinses (soak method), and the fruit rinse/broths (rinse method) were incubated for 24 h at 35C. The Bacteriological Analytical Manual (BAM) Salmonella culture method was followed thereafter. The soak method produced significantly greater numbers (P < 0.05) of positive test portions than did the rinse method for the analysis of mangoes (93 versus 12) and tomatoes (85 versus 34). The 3 broths were comparable for the recovery of Salmonella for both the soak and the rinse methods for mangoes. For tomatoes, there were no significant differences among the broths for the soak method, but BPW and UP broth were significantlymore productive (P < 0.05) than LAC broth by the rinse method. In the second phase, the relative effectiveness of LAC broth, BPW, and UP broth for the recovery of Salmonella from comminuted fruit was examined. Fruits were contaminated with single Salmonella serovars and aged for 4 days at 26C. Twenty 25 g test portions were preenriched in each of the following broths: BPW, LAC broth, and UP broth. The BAM Salmonella culture method was followed thereafter. For cantaloupes, significantly more (P < 0.05) Salmonella-positive test portions were recovered with UP broth (96 Salmonella-positive test portions) and BPW (87 Salmonella-positive test portions) than with LAC broth (57 Salmonella-positive test portions). For mangoes, BPW recovered an arithmetically larger number of Salmonella-positive test portions (27 Salmonella-positive test portions) than did either LAC broth (14 Salmonella-positive test portions) or UP broth (18 Salmonella-positive test portions). For tomatoes, there were no significant differences among the broths: BPW recovered 65 Salmonella-positive test portions, UP broth recovered 62 Salmonella-positive test portions, and LAC broth recovered 60 Salmonella-positive test portions. For the analysis of whole fruit, it is recommended that the soak method be used. For whole fruit analyzed with the soak method, UP broth should be used for tomatoes and BPW should be used formangoes. It is further recommended that UP broth be used for the analysis of comminuted cantaloupes and that BPW be used for the analysis of comminuted mangoes and tomatoes.

Purpose: The serine-threonine kinase mammalian target of rapamycin has emerged as a potential target for cancer therapy. Rapamycin and rapamycin analogs are undergoing clinical trials and have induced clinical responses in a subgroup of patients. Rapamycin has also been reported to enhance the efficacy of several cytotoxic agents. The aim of this study was to determine the nature of the interactions between rapamycin and chemotherapeutic agents used as first- and second-line agents against breast cancer. Experimental Design: We performed a multiple drug effect/combination index isobologram analysis in cells sensitive and resistant to rapamycin alone in vitro, and we evaluated the in vivo efficacy of combination therapy in a rapamycin-sensitive model. Results: In vitro, synergistic interactions were observed in combinations with paclitaxel, carboplatin, and vinorelbine. Additive effects were observed in combinations with doxorubicin and gemcitabine. Rapamycin dramatically enhanced paclitaxel- and carboplatin-induced apoptosis. This effect was sequence dependent and mediated at least partly through caspase activation. Furthermore, rapamycin enhanced chemosensitivity to paclitaxel and carboplatin in HER2/neu-overexpressing cells, suggesting a potential approach to these poorly behaving tumors. Cell lines that are resistant to the growth-inhibitory effect of rapamycin were also resistant to rapamycin-mediated chemosensitization. In vivo, rapamycin combined with paclitaxel resulted in a significant reduction in tumor volume compared with either agent alone in rapamycin-sensitive tumors. Conclusions: Rapamycin potentiates the cytotoxicity of selected chemotherapeutic agents in cell lines sensitive to the effects of rapamycin due to aberrations in the phosphatidylinositol 3′-kinase/Akt pathway, suggesting that combination therapy may be effective in patients selected for aberrations in this pathway.

Alternative methods of reducing oxygen during anaerobic enrichment in the Bacteriological Analytical Manual (BAM) Shigella culture method were evaluated and compared to the current and less practical GasPak® method. The alternative anaerobic methods included the use of reducing agents in Shigella broth and reducing culture container headspace volume to minimize atmospheric effects on oxygen concentration in Shigella broth during enrichment. The reducing agents evaluated were sodium thioglycollate, L-cystine, L-cysteine, titanium(III) citrate, and dithiothreitol, each at concentrations of 0.1, 0.05, and 0.01%. The use of Oxyrase for Broth® with the enrichment medium (Shigella broth) was evaluated at concentrations of 10, 20 and 30 μL/mL. Recoveries of chill- and freeze-stressed S. sonnei strains 357 and 20143 were determined with each anaerobic method, including the GasPak method, using inoculation levels ranging from 100 to 103 cells. For each anaerobic method, strain, inoculation level, and stress type, 5 replicate enrichments were evaluated by streaking to MacConkey agar for isolation. The numbers of cultures with each method from which S. sonnei was isolated were used to compare the alternative anaerobic methods to the GasPak method. The alternative anaerobic method with which chill- and freeze-stressed S. sonnei strains 357 and 20143 were isolated most consistently was the use of Oxyrase for Broth in Shigella broth at a concentration of 20 μL/mL. This method was compared to the GasPak anaerobic method in evaluations on the recovery of S. sonnei strains 357 and 20143 from artificially contaminated test portions of parsley, cilantro, green onions, strawberries, carrots, and celery. A third anaerobic method included the use of 0.5 cm mineral oil overlay on cultures containing Oxyrase for Broth at concentrations of 20 μL/mL. Recovery rates of strain 357 were significantly greater (p < 0.05) with the GasPak method than with Oxyrase for Broth, with and without the 0.5 cm mineral oil overlay, for test portions of parsley, cilantro, and celery. When Oxyrase for Broth was used with Shigella broth, strain 357 was isolated at higher rates from all produce types, except cilantro, when 0.5 cm mineral oil overlay was applied to enrichment cultures. The use of mineral oil overlay with Oxyrase for Broth also improved recovery of strain 20143 from test portions of all produce types except green onion and strawberries. These differences were significant (p < 0.05) with parsley, carrots, and cilantro (1 of 2 evaluations). No statistically significant differences (p > 0.05) between the GasPak and Oxyrase for Broth anaerobic methods occurred when mineral oil overlay was used with Oxyrase for Broth. The use of Oxyrase for Broth with a 0.5 cm mineral oil overlay is a practical alternative for anaerobic enrichment with the BAM method in the analysis of some produce types. Differences in recovery among the different produce types and methods occurred between S. sonnei strains 357 and 20143, emphasizing the need for additional S. sonnei strains in future evaluations.

Soak and rinse methods were compared for the recovery of Salmonella from whole cantaloupes. Cantaloupes were surface inoculated with Salmonella cell suspensions and stored for 4 days at 2 to 6 degrees C. Cantaloupes were placed in sterile plastic bags with a nonselective preenrichment broth at a 1:1.5 cantaloupe weight-to-broth volume ratio. The cantaloupe broths were shaken for 5 min at 100 rpm after which 25-ml aliquots (rinse) were removed from the bags. The 25-ml rinses were preenriched in 225-ml portions of the same uninoculated broth type at 35 degrees C for 24 h (rinse method). The remaining cantaloupe broths were incubated at 35 degrees C for 24 h (soak method). The preenrichment broths used were buffered peptone water (BPW), modified BPW, lactose (LAC) broth, and Universal Preenrichment (UP) broth. The Bacteriological Analytical Manual Salmonella culture method was compared with the following rapid methods: the TECRA Unique Salmonella method, the VIDAS ICS/SLM method, and the VIDAS SLM method. The soak method detected significantly more Salmonella-positive cantaloupes (P < 0.05) than did the rinse method: 367 Salmonella-positive cantaloupes of 540 test cantaloupes by the soak method and 24 Salmonella-positive cantaloupes of 540 test cantaloupes by the rinse method. Overall, BPW, LAC, and UP broths were equivalent for the recovery of Salmonella from cantaloupes. Both the VIDAS ICS/SLM and TECRA Unique Salmonella methods detected significantly fewer Salmonella-positive cantaloupes than did the culture method: the VIDAS ICS/SLM method detected 23 of 50 Salmonella-positive cantaloupes (60 tested) and the TECRA Unique Salmonella method detected 16 of 29 Salmonella-positive cantaloupes (60 tested). The VIDAS SLM and culture methods were equivalent: both methods detected 37 of 37 Salmonella-positive cantaloupes (60 tested).

The relative efficiencies of 2 Bacteriological Analytical Manual (BAM) pre-enrichments, lactose broth (LAC) and brilliant green water (BGW), were compared with Universal Pre-enrichment (UP) broth for the recovery of individual Salmonella serovars from instant nonfat dry milk (NFDM), dry whole milk (DWM), lactic casein (LC), and liquid whole milk (LWM). BGW was compared with UP broth for the analysis of NFDM and DWM but not with the other 2 matrixes. LAC was compared with UP broth for the analysis of LC and LWM. UP broth was made both from a commercial dehydrated preparation (UPC) and from individual ingredients (UPI). Bulk quantities of the selected dairy foods were inoculated with Salmonella serovars at levels intended to produce fractionally positive results, where at least half of the test portions analyzed, with one of the methods being evaluated, would be shown to be Salmonella-positive. For NFDM, in 6 of 9 experiments, with 2 different Salmonella serovars, BGW was significantly more productive than either UPI or UPC broth (p < 0.05). Salmonella was recovered from 118 of 180 test portions with BGW, from 25 of 180 test portions with UPC, and from 14 of 180 test portions with UPI. For DWM, in 2 of 4 experiments, with 2 different Salmonella serovars, BGW was significantly more productive than either UPI or UPC broth (p < 0.05). Salmonella was recovered from 67 of 80 test portions with BGW, from 36 of 80 test portions with UPC, and from 37 of 80 test portions with UPI. For LWM, in 9 of 9 experiments, with 3 different Salmonella serovars, there were no significant differences among the broths. Salmonella was recovered from 120 of 180 test portions with LAC, from 135 of 180 test portions with UPC, and from 129 of 180 test portions with UPI. For LC, in 5 of 7 experiments, with 2 different Salmonella serovars, both UPI and UPC broth were significantly more productive than LAC (p < 0.05). Salmonella was recovered from 42 of 140 test portions with LAC, from 114 of 140 test portions with UPC, and from 114 of 140 test portions with UPI. In addition, overall results showed that UPC and UPI broths were equivalent for the recovery of Salmonella from the foods tested, without regard to their performance in comparison with either LAC or BGW.

Responding to a need for a guide for conducting Official Method validation studies of microbiological methods, AOAC utilized the experience of three microbiologists who have been active in the field of method validation. In collaboration, a document was prepared which covered the following areas: terms and their definitions associated with the Official Methods program (e.g., reference methods, alternative methods, and ruggedness testing), protocols and validation requirements for qualitative methods versus those for quantitative methods, the concept of the precollaborative study, ruggedness testing, tests for significant differences, performance indicators, and the approval process. After its preparation, this document was reviewed by the members of the Methods Committee on Microbiology and Extraneous Materials and by members of the Official Methods Board. Herein is presented the approved version of that document.

A comparison was made of the relative efficiencies of Universal Preenrichment (UP) broth and lactose broth for the recovery of a variety of Salmonella serovars from pasteurized and unpasteurized apple cider and pasteurized apple juice. Bulk portions of juice were contaminated with single Salmonella serovars at high and low levels of 0.4 and 0.04 CFU/mL, respectively. The juice was aged for a minimum of 5 days at 2–5°C. On the day analysis was initiated, each of 20 test portions (25 mL) of the contaminated juice was preenriched in UP broth and in lactose broth. The Bacteriological Analytical Manual Salmonella culture method was followed thereafter. For pasteurized apple cider, UP broth recovered significantly (p < 0.05) more Salmonella-positive test portions than did lactose broth (112 and 75, respectively). For unpasteurized apple cider, UP broth recovered significantly more Salmonella-positive test portions than did lactose broth (326 and 221, respectively). For pasteurized apple juice, UP broth recovered more Salmonella-positive test portions than did lactose broth (93 and 81, respectively). However, this difference was not statistically significant. These results indicate that UP broth should replace lactose broth for the analysis of pasteurized and unpasteurized apple cider and pasteurized apple juice.

The aim of this study is to review the psychotherapeutic intervention models used in the elderly who have "fear of falling" and to identify the results. The literature search was performed on the basis of MEDLINE, LILACS and SciELO and ten studies with different designs and interventions were accessed, including two review articles about the fear of falling in the elderly. Among the eight articles that showed intervention models, seven were based on the cognitive-behavioral therapy (CBT) and one was not ​​clear about its driving line. The studies that showed better results in clinical practice used the multidisciplinary intervention. These two multidisciplinary studies used psychology and physiotherapy for assessment and treatment for fear of falling in the elderly, with good effect on health after a six-month-follow-up. Although it presents itself as a public health issue, with high prevalence in the elderly with fear of falling, preceded by a fall or not, there is a lack of studies on models for intervention and recovery of possible impacts that influence routine from elderly with fear of falling. The main finding of this study was the characterization of the fear of falling as a phobia, thus connected to an anxiety disorder, which helps to guide treatment, prognosis and plan of action of the professionals involved. It was concluded that a review of existing models for the fear of falling must be expanded worldwide and that there should be guided research on the models with the best results. This maturing and structuring of knowledge in the field of elderly psyche can come through greater involvement of psychologists in research, adjustments and tests of models for the fear of falling, as well as other specific frames and for the initial evaluation, with the elderly population.

Twenty-three laboratories participated in a collaborative study to compare the relative effectiveness of Rappaport-Vassiliadis (RV) medium incubated at 42 degrees C, selenite cystine (SC) broth (35 degrees C), and tetrathionate (TT) broth (35 and 43 degrees C) for recovery of Salmonella from the following foods with a low microbial load: dried egg yolk, dry active yeast, ground black pepper, guar gum, and instant nonfat dry milk. For dry active yeast, lauryl tryptose (LT) broth, incubated at 35 degrees C, was used instead of SC broth. All of the foods were artificially inoculated with single Salmonella serovars, that had been lyophilized before inoculation, at high and low target levels of 0.4 and 0.04 colony forming units/g food, respectively. For analysis of 870 test portions, representing all of the foods except yeast, 249 Salmonella-positive test portions were detected by RV medium, 265 by TT broth (43 degrees C), 268 by TT broth (35 degrees C), and 269 by SC broth (35 degrees C). For analysis of 225 test portions of yeast, 79 Salmonella-positive test portions were detected by RV medium, 79 by TT broth (43 degrees C), 84 by TT broth (35 degrees C), and 68 by LT broth (35 degrees C). RV medium was comparable to, or even more effective than, the other selective enrichments for recovery of Salmonella from all of the foods except guar gum. It is recommended that RV (42 degrees C) and TT (35 degrees C) be used with foods that have a low microbial load, except for guar gum for which SC (35 degrees C) and TT (35 degrees C) are recommended.