Rapid identification of Salmonella serovars Enteritidis and Typhimurium using whole cell matrix assisted laser desorption ionization - Time of flight mass spectrometry (MALDI-TOF MS) coupled with multivariate analysis and artificial intelligence.

Salmonella is a common food-borne pathogen with Enteritidis and Typhimurium being among the most important serovars causing numerous outbreaks. A rapid method was investigated to identify these serovars using whole-cell MALDI-TOF MS coupled with multivariate analysis and artificial intelligence and 113 Salmonella strains, including 38 Enteritidis (SE), 38 Typhimurium (ST) and 37 strains from 32 other Salmonella serovars (SG). Datasets of ions (presence/absence) with high discriminative power were created using newly developed criteria and subject to multivariate analyses and eight artificial intelligence (AI) tools. Principal Component Analysis based on 55 or 88 selected ions separated SE, ST and SG without overlap on the first three principal components. Datasets were partitioned using five partitioning methods with 70% of samples for AI model training and 30% for validation. Of the eight AI models evaluated, high performance (HP) SVM and HP Neural were the top performers, identified three serovar groups 97% correctly on average (range 82%-100%) according to the validation results. Selection of serovar specific ions facilitated differentiation of serotypes using unsupervised model PCA and improved the accuracy of classification using AI significantly (p < 0.01). MALDI-TOF MS incorporated with advanced data processing and classification tools is a promising method to allow rapid identification of Salmonella serovars of concern in routine diagnostic laboratories.

Application of a CRISPR Sequence-Based Method for a Large-Scale Assessment of Salmonella Serovars in Ontario Poultry Production Environments.

Accurate detection of all Salmonella serovars present in a sample is important in surveillance programs. Current detection protocols are limited to detection of a predominant serovar, missing identification of less abundant serovars in a sample. An alternative method, called CRISPR-SeroSeq, serotyping by sequencing of amplified CRISPR spacers, was employed to detect multiple serovars in a sample without the need of culture isolation. The CRISPR-SeroSeq method successfully detected 34 most frequently reported Salmonella serovars in pure cultures and target serovars at 104 CFU/mL in 27 Salmonella-negative environmental enrichment samples post-spiked with one of 15 different serovars, plus 2 additional serovars at 1 log CFU/mL higher abundance. When the method was applied to 442 naturally contaminated environmental samples collected from 192 poultry farms, 25 different serovars were detected from 430 of the samples. In 73.1% of the samples, 2 to 7 serovars were detected, with Salmonella Kiambu (55.7%), Salmonella Infantis (48.4%), Salmonella Kentucky (27.1%), Salmonella Livingstone (26.6%), and Salmonella Mbandaka/Montevideo (23.4%) being the most prevalent on the farms. Single isolates from 384 samples were also analyzed using a traditional serotyping method, and the same serovar identified by culture was detected by CRISPR-SeroSeq in 96.1% (369/384) of samples, with the former missing detection of additional and sometimes critical serovars. The surveillance data obtained via CRISPR-SeroSeq revealed a significant emergence of Salmonella Kiambu and Salmonella Rissen on poultry farms in Ontario. The results highlight the effectiveness of the CRISPR-SeroSeq approach in detecting multiple Salmonella serovars in poultry environmental samples under applied conditions, providing updated surveillance information on Salmonella serovars on poultry farms in Ontario. IMPORTANCE The CRISPR-SeroSeq method represents an alternative molecular tool to the traditional culture-based serotyping method that can detect multiple Salmonella serovars in a sample and provide rapid serovar results without the need of selective enrichment and culture isolation. The evaluation results can facilitate implementation of the method in routine Salmonella surveillance on poultry farms and in outbreak investigations. The application of the method can increase the accuracy of current serovar prevalence information. The results highlight the effectiveness of the validated method and the need for monitoring Salmonella serovars in poultry environments to improve current surveillance programs. The updated surveillance data provide timely information on emergence of different Salmonella serovars on poultry farms in Ontario and support on-farm risk assessment and risk management of Salmonella.

How, Why and When Tsallis Statistical Mechanics Provides Precise Descriptions of Natural Phenomena.

The limit of validity of ordinary statistical mechanics and the pertinence of Tsallis statistics beyond it is explained considering the most probable evolution of complex systems processes. To this purpose we employ a dissipative Landau-Ginzburg kinetic equation that becomes a generic one-dimensional nonlinear iteration map for discrete time. We focus on the Renormalization Group (RG) fixed-point maps for the three routes to chaos. We show that all fixed-point maps and their trajectories have analytic closed-form expressions, not only (as known) for the intermittency route to chaos but also for the period-doubling and the quasiperiodic routes. These expressions have the form of q-exponentials, while the kinetic equation's Lyapunov function becomes the Tsallis entropy. That is, all processes described by the evolution of the fixed-point trajectories are accompanied by the monotonic progress of the Tsallis entropy. In all cases the action of the fixed-point map attractor imposes a severe impediment to access the system's built-in configurations, leaving only a subset of vanishing measure available. Only those attractors that remain chaotic have ineffective configuration set reduction and display ordinary statistical mechanics. Finally, we provide a brief description of complex system research subjects that illustrates the applicability of our approach.

Number theory, borderline dimension and extensive entropy in distributions of ranked data.

The consideration of an existing stochastic approach for the reproduction of ranked data pointed at a formal equivalence between its key mathematical expression and that for trajectories at the tangent bifurcation. This fact led to a nonlinear dynamical approach for rank distributions that shows similarities with universality classes in critical phenomena. The renormalization group (RG) fixed-point map f*(x) for a tangent bifurcation of arbitrary nonlinearity z > 1 has proved to be a powerful tool into which the formalism can be couched. The source distribution P(N) of the stochastic approach can be linked to f*(x) while the size-rank N(k) and frequency-rank F(k') distributions are obtained, respectively, from the map trajectories xt and the sums of its positions. We provide now an extension to Number Theory as we obtain from the trajectories xt of f*(x) the numbers, or asymptotic approximations of them, for the Factorial, Natural, Prime and Fibonacci sets. A measure of the advance of these numbers towards infinity is given by sums of positions that represent their reciprocals. We specify rank distribution universality classes, already associated with real data, to these number sets. We find that the convergence of the series of number reciprocals occurs first at nonlinearity z = 2, that which corresponds to the classical Zipf law, and link this transition edge to the action of the attractor when it first reduces the fractal dimension of trajectory positions to zero. Furthermore, the search of logarithmic corrections common to borderline dimensions provides a link to the Prime numbers set. Finally, we find corroborating evidence of these logarithmic corrections from the analysis of large data sets for ranked earthquake magnitudes. The formalism links all types of ranked distributions to a generalized extensive entropy.

Distribution of human papillomavirus genotypes by severity of cervical lesions in HPV screened positive women from the ESTAMPA study in Latin America.

The proportion of HPV16 and 18-associated cervical cancer (CC) appears rather constant worldwide (≥70%), but the relative importance of the other HR-HPV differs slightly by geographical region. Here, we studied the HPV genotype distribution of HPV positive Latin American (LA) women by histological grade, in a sub-cohort from the ESTAMPA study; we also explored the association of age-specific HPV genotypes in severe lesions. Cervical samples from 1,252 participants (854 ≤CIN1, 121 CIN2, 194 CIN3 and 83 CC) were genotyped by two PCRs-Reverse Blotting Hybridization strategies: i) Broad-Spectrum General Primers 5+/6+ and ii) PGMY9/11 PCRs. HPV16 was the most frequently found genotype in all histological grades, and increased with the severity of lesions from 14.5% in ≤ CIN1, 19.8% in CIN2, 51.5% in CIN3 to 65.1% in CC (p < 0.001). For the remaining HR-HPVs their frequency in CC did not increase when compared to less severe categories. The nonavalent vaccine HR-types ranked at the top in CC, the dominant ones being HPV16 and HPV45. HR-HPV single infection occurs, respectively, in 57.1% and 57.0% of ≤CIN1 and CIN2, increasing to 72.2% and 91.6% in CIN3 and CC (p<0.001). No association between age and HPV type was observed in CC, although the risk of HPV16 infection in CIN3 cases increased with age. Results confirm the relevance of HPV16 in the whole clinical spectrum, with a strong rise of its proportion in CIN3 and cancer. This information will be relevant in evaluating the impact of HPV vaccination, as a baseline against which to compare genotype changes in HPV type-specific distribution as vaccinated women participate in screening in LA region. Likewise, these data may help select the best HPV testing system for HPV-based efficient, affordable, and sustainable screening programmes.

Antiviral activity of the sea cucumber tegument extract (Pattalus mollis) on human rotavirus A (RVA).

The antiviral effect against RVA in cell culture was evaluated by using an aqueous extract of Patallus mollis sea cucumber, applying the titration methodology. This technique is used to measures the ability of the extract dilutions to inhibit the cytopathic effect (CPE) of the virus, expressed as percentage of inhibition (IP). The mean extract cytotoxic concentration (CC50) used in the antiviral assay was 27,042.10 µg/mL and the PI of the antiviral activity extract was greater than 99.9% for each concentration. To determine the viral action mode, the cells were previously treated with the extracts in different stages during the viral infection cycle. The result analysis suggests that the extract inhibits 99% of the virus during the absorption and viral inactivation phase. These results show the P. mollis extract has a remarkable antiviral effect against the RVA in cell culture. So that, it is crucial to investigate its action mechanisms.

Yersinia Phages and Food Safety.

One of the human- and animal-pathogenic species in genus Yersinia is Yersinia enterocolitica, a food-borne zoonotic pathogen that causes enteric infections, mesenteric lymphadenitis, and sometimes sequelae such as reactive arthritis and erythema nodosum. Y. enterocolitica is able to proliferate at 4 C, making it dangerous if contaminated food products are stored under refrigeration. The most common source of Y. enterocolitica is raw pork meat. Microbiological detection of the bacteria from food products is hampered by its slow growth rate as other bacteria overgrow it. Bacteriophages can be exploited in several ways to increase food safety with regards to contamination by Y. enterocolitica. For example, Yersinia phages could be useful in keeping the contamination of food products under control, or, alternatively, the specificity of the phages could be exploited in developing rapid and sensitive diagnostic tools for the identification of the bacteria in food products. In this review, we will discuss the present state of the research on these topics.

Dynamical analogues of rank distributions.

We present an equivalence between stochastic and deterministic variable approaches to represent ranked data and find the expressions obtained to be suggestive of statistical-mechanical meanings. We first reproduce size-rank distributions N(k) from real data sets by straightforward considerations based on the assumed knowledge of the background probability distribution P(N) that generates samples of random variable values similar to real data. The choice of different functional expressions for P(N): power law, exponential, Gaussian, etc., leads to different classes of distributions N(k) for which we find examples in nature. Then we show that all of these types of functions can be alternatively obtained from deterministic dynamical systems. These correspond to one-dimensional nonlinear iterated maps near a tangent bifurcation whose trajectories are proved to be precise analogues of the N(k). We provide explicit expressions for the maps and their trajectories and find they operate under conditions of vanishing or small Lyapunov exponent, therefore at or near a transition to or out of chaos. We give explicit examples ranging from exponential to logarithmic behavior, including Zipf's law. Adoption of the nonlinear map as the formalism central character is a useful viewpoint, as variation of its few parameters, that modify its tangency property, translate into the different classes for N(k).

Decontamination of Chia and Flax Seed Inoculated with Salmonella and Surrogate, Enterococcus faecium NRRL B-2354, Using a Peracetic Acid Sanitizing Solution: Antimicrobial Efficacy and Impact on Seed Functionality.

Raw chia and flax seeds are increasingly associated with Salmonella contamination. However, intervention technologies for these seeds that maintain them in a raw state, without causing clumping because of mucilage production upon moisture exposure, are limited. In this study, a commercial ethanol and paracetic acid sanitizing solution meeting these criteria was evaluated for efficacy against Salmonella and Enterococcus faecium NRRL B-2354, a known Salmonella surrogate for thermal intervention technologies. Samples (100 g each) of chia and flax seeds ( n = 5) were inoculated with either a cocktail of Salmonella Newport, Senftenberg, Oranienburg, Saintpaul, Typhimurium DT104, and Cubana or E. faecium NRRL B-2354. After overnight acclimatization, samples were treated with 4 mL of sanitizing solution per sample and then held at ambient temperature (20 to 25°C) for 1 h before bacterial enumeration. Separate 1-kg-treated batches were evaluated for germination ability (4 replicates of 100-g samples), as well as nutrient content and rancidity ( n = 3), compared with untreated control. Following the posttreatment holding time, these batches were dried back to original moisture content at 70°C to evaporate residual sanitizing solution, thereby stopping treatment. The sanitizing solution was found to be an effective intervention method for chia and flax seeds, reducing Salmonella to below the level of detection by more than 4 and more than 5 average log CFU/g, respectively. Germination was not significantly affected ( P ≥ 0.05) for chia seed. For both seeds, nutrition and rancidity were not significantly affected ( P ≥ 0.05). Furthermore, E. faecium NRRL B-2354 was found to be an appropriate Salmonella surrogate for treatment of chia and flax seeds with this sanitizing solution, showing comparable but higher resistance to treatment with the sanitizing solution than the Salmonella cocktail.

Draft Genome Sequences of Two Novel Salmonella enterica subsp. enterica Strains Isolated from Low-Moisture Foods with Applications in Food Safety Research.

The genomes of two strains of Salmonella enterica subsp. enterica serovar Cubana and serovar Muenchen, isolated from dry hazelnuts and chia seeds, respectively, were sequenced using the Illumina MiSeq platform, assembled de novo using the overlap-layout-consensus method, and aligned to their respective most identical sequence genome scaffolds using MUMMER and BLAST searches.

Rank distributions: Frequency vs. magnitude.

We examine the relationship between two different types of ranked data, frequencies and magnitudes. We consider data that can be sorted out either way, through numbers of occurrences or size of the measures, as it is the case, say, of moon craters, earthquakes, billionaires, etc. We indicate that these two types of distributions are functional inverses of each other, and specify this link, first in terms of the assumed parent probability distribution that generates the data samples, and then in terms of an analog (deterministic) nonlinear iterated map that reproduces them. For the particular case of hyperbolic decay with rank the distributions are identical, that is, the classical Zipf plot, a pure power law. But their difference is largest when one displays logarithmic decay and its counterpart shows the inverse exponential decay, as it is the case of Benford law, or viceversa. For all intermediate decay rates generic differences appear not only between the power-law exponents for the midway rank decline but also for small and large rank. We extend the theoretical framework to include thermodynamic and statistical-mechanical concepts, such as entropies and configuration.

A Study To Assess the Numbers and Prevalence of Bacillus cereus and Its Toxins in Pasteurized Fluid Milk.

Bacillus cereus is a pathogenic adulterant of raw milk and can persist as spores and grow in pasteurized milk. The objective of this study was to determine the prevalence of B. cereus and its enterotoxins in pasteurized milk at its best-before date when stored at 4, 7, and 10°C. More than 5.5% of moderately temperature-abused products (stored at 7°C) were found to contain >105 CFU/mL B. cereus , and about 4% of them contained enterotoxins at a level that may result in foodborne illness; in addition, more than 31% of the products contained >105 CFU/mL B. cereus and associated enterotoxins when stored at 10°C. Results from a growth kinetic study demonstrated that enterotoxin production by B. cereus in pasteurized milk can occur in as short as 7 to 8 days of storage at 7°C. The higher B. cereus counts were associated with products containing higher butterfat content or with those produced using the conventional high-temperature, short-time pasteurization process. Traditional indicators, aerobic colony counts and psychrotrophic counts, were found to have no correlation with level of B. cereus in milk. The characterization of 17 representative B. cereus isolates from pasteurized milk revealed five toxigenic gene patterns, with all the strains carrying genes encoding for diarrheal toxins but not for an emetic toxin, and with one strain containing all four diarrheal enterotoxin genes (nheA, entFM, hblC, and cytK). The results of this study demonstrate the risks associated even with moderately temperature-abused pasteurized milk and the necessity of a controlled cold chain throughout the shelf life of fluid milk to enhance product safety and minimize foodborne illness.

Recurrencia/persistencia de neoplasia intraepitelial cervical posterior a cono LEEP en el Instituto Nacional de Enfermedades Neoplásicas / Recurrence/persistence of cervical intraepithelial neoplasia after undergoing a LEEP conization at Instituto Nacional de Enfermedades Neoplásicas

Objetivo: Determinar la incidencia de recurrencia/persistencia de neoplasia intraepitelial cervical (NIC) en pacientes sometidos a procedimiento de escisión electroquirúrgica en asa (cono LEEP) en el Servicio de Ginecología Oncológica del Instituto Nacional de Enfermedades Neoplásicas (INEN). Materiales y métodos: Estudio de cohorte, retrospectivo en el cual se revisaron las historias clínicas de pacientes sometidos a cono LEEP durante el periodo de enero del 2002 a diciembre del 2003. Los pacientes fueron divididos en un grupo con margen comprometido y otro con margen negativo después del procedimiento de cono LEEP. El seguimiento de los pacientes se realizó cada 3 meses con examen físico, PAP y colposcopía. El desenlace fue la recurrencia de NIC. Para valorar el riesgo de recurrencia de NIC entre los grupos de estudio se calculó el riesgo relativo (RR) con su respectivo intervalo de confianza (IC) al 95%. Resultados: En total 451 pacientes fueron sometidos a cono LEEP durante el periodo de estudio. La edad promedio fue 35.32 años, el promedio de parejas sexuales fue 2.1 y el promedio de hijos fue 2.7. Presentaron margen comprometido 109 (25%) y margen negativo 342 (75%). De ellos solo 94 y 102, respectivamente, cumplían los criterios de elegibilidad y fueron seguidos. La media de seguimiento fue de 21.74 meses. La incidencia de recurrencia/persistencia dentro de los pacientes con margen comprometido fue 5.5% mientras que la incidencia de recurrencia en un grupo de pacientes con margen negativo fue 1.96% No encontramos diferencias significativas en el riesgo de padecer recurrencia de NIC en el grupo de pacientes con margen comprometido versus los pacientes con margen negativo. RR= 2.7; IC 95% (0.41 a 33.8). Conclusión: No se encontró diferencia significativa en la incidencia de neoplasia intracervical para pacientes con margen comprometido y los pacientes con margen negativo.

Objective: To determine the incidence of recurrence/persistence of cervical intraepithelial neoplasia (CIN) in patients who underwent a loop electrosurgical excision procedure (LEEP) at the Department of Gynecologic Oncology of Instituto Nacional de Enfermedades Neoplásicas (INEN). Materials and methods: A retrospective cohort study in which medical records of patients who underwent a LEEP conization from January 2002 to December 2003 were examined. After the LEEP conization, patients were divided into two groups: one with positive margin and another one with negative margin. Follow-up was performed every 3 months and consisted of a physical examination, Pap test and colposcopy. The outcome was recurrence of CIN. To assess the risk of CIN recurrence among the study groups, relative risk (RR) and its respective 95% confidence interval (CI) were calculated. Results: A total of 451 patients underwent a LEEP conization during the study period. The mean age was 35.32 years old, the mean number of sexual partners was 2.1 and the mean number of children was 2.7. One hundred and nine (109) patients (25%) showed a positive margin and 342 (75%) showed a negative one. Out of these subjects, only 94 and 102, respectively, met the eligibility criteria and were followed. The mean follow-up period was 21.74 months. The incidence of recurrence/persistence in patients with positive margin was 5.5%, while the incidence of recurrence/persistence in the group of patients with negative margin was 1.96%. We found no significant differences in the risk of CIN recurrence in the group of patients with positive margin versus patients with negative margin. RR = 2.7, 95% CI (0.41 to 33.8). Conclusion: There was no significant difference in the incidence of cervical intraepithelial neoplasia between patients with positive margin and patients with negative margin.

Actividad antiinflamatoria in vitro de los polisacáridos sulfatados de Patallus mollis extraídos mediante digestión enzimática / In vitro anti-inflammatory effect of Patallus mollis sulphated polysaccharides extracted by enzymatic digestion

Objetivo: Evaluar la actividad antiinflamatoria in vitro de los polisacáridos (PCs) del tegumento de Patallus mollis mediante la prueba de la estabilidad de la membrana celular del glóbulo rojo y la desnaturalización de la albumina. Materiales y métodos: Los especímenes fueron recolectados en la bahía de Pucusana, se identificaron y los PCs se extrajeron mediante digestión enzimática con bromelina (PBr), papaína (PPa) y pepsina (PPe). Se cuantifico el contenido de azucares, sulfatos y proteínas totales mediante métodos colorimétricos. Para la prueba de estabilidad de la membrana, se preparó una solución de glóbulos rojos [10 % v/v], la cual se mezcló con PBS, NaCl [0,36%], y PCs [2000 ­ 62.5 µg/mL]. Para la prueba de la desnaturalización de la albúmina, la mezcla consistió en suero de albúmina bovina [0,5%] con PCs [2000 ­ 62.5 µg/mL]. Como controles se utilizó diclofenaco sódico y agua destilada. El análisis de resultados fue en base a la absorbancia medida en un espectrofotómetro de placas a 420 y 660 nm, según las pruebas respectivamente. Resultados: Los PCs presentaron niveles elevados de azúcares y proteínas totales, así como presencia de sulfatos totales. Además los PCs exhibieron una capacidad estabilizadora de la membrana, siendo la máxima presentada por PBr a una concentración de 62.5 µg/mL. Solo el PPa presentó inhibición de la desnaturalización de albúmina inducida por calor a una dosis de 2000 µg/mL. Conclusión: Nuestros resultados sugieren que los PCs tiene actividades antiinflamatorias y son candidatos potenciales para la formulación de un ingrediente alimentario funcional y / o fármaco para tratar enfermedades inflamatorias.

Propagation method for persistent high yield of diverse Listeria phages on permissive hosts at refrigeration temperatures.

The efficient production of a high concentration of bacteriophage in large volumes has been a limiting factor in the exploration of the true potential of these organisms for biotechnology, agriculture and medicine. Traditional methods focus on generating small volumes of highly concentrated samples as the end product of extensive mechanical and osmotic processing. To function at an industrial scale mandates extensive investment in infrastructure and input materials not feasible for many smaller facilities. To address this, we developed a novel, scalable, generic method for producing significantly higher titer psychrophilic phage (P < 2.0 × 10(-6)), 2- to 4-fold faster than traditional methods. We generate renewable high yields from single source cultures by propagating phage under refrigeration conditions in which Listeria, Yersinia and their phages grow in equilibrium. Diverse Yersinia and Listeria phages tested yielded averages of 3.49 × 10(8) to 3.36 × 10(12) PFU/ml/day compared to averages of 1.28 × 10(5) to 1.30 × 10(10) PFU/ml/day by traditional methods. Host growth and death kinetics made this method ineffective for extended propagation of mesophilic phages.

Yersinia enterocolitica-Specific Infection by Bacteriophages TG1 and ϕR1-RT Is Dependent on Temperature-Regulated Expression of the Phage Host Receptor OmpF.

UNLABELLED: Bacteriophages present huge potential both as a resource for developing novel tools for bacterial diagnostics and for use in phage therapy. This potential is also valid for bacteriophages specific for Yersinia enterocolitica To increase our knowledge of Y. enterocolitica-specific phages, we characterized two novel yersiniophages. The genomes of the bacteriophages vB_YenM_TG1 (TG1) and vB_YenM_ϕR1-RT (ϕR1-RT), isolated from pig manure in Canada and from sewage in Finland, consist of linear double-stranded DNA of 162,101 and 168,809 bp, respectively. Their genomes comprise 262 putative coding sequences and 4 tRNA genes and share 91% overall nucleotide identity. Based on phylogenetic analyses of their whole-genome sequences and large terminase subunit protein sequences, a genus named Tg1virus within the family Myoviridae is proposed, with TG1 and ϕR1-RT (R1RT in the ICTV database) as member species. These bacteriophages exhibit a host range restricted to Y. enterocolitica and display lytic activity against the epidemiologically significant serotypes O:3, O:5,27, and O:9 at and below 25°C. Adsorption analyses of lipopolysaccharide (LPS) and OmpF mutants demonstrate that these phages use both the LPS inner core heptosyl residues and the outer membrane protein OmpF as phage receptors. Based on RNA sequencing and quantitative proteomics, we also demonstrate that temperature-dependent infection is due to strong repression of OmpF at 37°C. In addition, ϕR1-RT was shown to be able to enter into a pseudolysogenic state. Together, this work provides further insight into phage-host cell interactions by highlighting the importance of understanding underlying factors which may affect the abundance of phage host receptors on the cell surface. IMPORTANCE: Only a small number of bacteriophages infecting Y. enterocolitica, the predominant causative agent of yersiniosis, have been previously described. Here, two newly isolated Y. enterocolitica phages were studied in detail, with the aim of elucidating the host cell receptors required for infection. Our research further expands the repertoire of phages available for consideration as potential antimicrobial agents or as diagnostic tools for this important bacterial pathogen.

Method Modification Study for the Thermo Scientific SureTect™ Listeria Species Assay-Matrix Extension.

The Thermo Scientific™ SureTect™ Listeria species assay is a new real-time PCR assay for the detection of all species of Listeria in food and environmental samples. The assay was originally certified as Performance Tested Methods(SM) (PTM) 071304 in 2013. This report details the method modification study undertaken to extend the performance claims of the assay for matrixes of raw ground turkey, raw ground pork, bagged lettuce, raw pork sausages, pasteurized 2% fat milk, raw cod, pasteurized brie cheese, and ice cream. The method modification study was conducted using the AOAC Research Institute (RI) PTM program to validate the SureTect PCR assay in comparison to the reference method detailed in ISO 11290-1:1996 including amendment 1:2004. All matrixes were tested by Thermo Fisher Scientific (Basingstoke, United Kingdom). In addition, three matrixes (raw cod, bagged lettuce, and pasteurized brie cheese) were analyzed independently as part of the AOAC RI-controlled independent laboratory study by the University of Guelph, Canada. Using probability of detection statistical analysis, there was no significant difference in the performance between the SureTect assay and the International Organization for Standardization reference method for any of the matrixes analyzed in this study.

Method Modification of the Thermo Scientific SureTect Listeria monocytogenes Assay for Raw Meat, Dairy, Produce, and Seafood.

The Thermo Scientific™ SureTect™ Listeria monocytogenes assay is a real-time PCR assay for the detection of Listeria monocytogenes in food and environmental samples, which was certified during 2013 by the AOAC Research Institute (RI) as Performance Tested Method(SM) (PTM) 061302 for a representative range of key food matrixes and production surfaces. This report details the method modification study, which was conducted during 2014, using the AOAC-RI PTM program to extend the validated matrix claims of the assay in comparison to the reference method detailed in International Organization for Standardization 11290-1:1996, including Amendment 1:2004, to gain certification for raw ground turkey, raw ground pork, pasteurized 2% milk, raw pork sausages, raw cod, pasteurized brie cheese, cooked sliced ham, and bagged lettuce. All matrixes were tested by Thermo Fisher Scientific, Microbiology Division, Basingstoke, UK. In addition, brie cheese, bagged lettuce, and raw cod were analyzed independently by the University of Guelph, Canada, during the AOAC-RI controlled independent laboratory study. Using probability of detection (POD) statistical analysis, a significant difference was demonstrated between the candidate and reference methods for the high spiking level with raw ground pork and brie cheese. For all other matrixes and the low spiked levels for raw ground pork and brie cheese, no significant difference by POD was seen between the two methods during the study.

Entropies for severely contracted configuration space.

We demonstrate that dual entropy expressions of the Tsallis type apply naturally to statistical-mechanical systems that experience an exceptional contraction of their configuration space. The entropic index [Formula: see text] describes the contraction process, while the dual index [Formula: see text] defines the contraction dimension at which extensivity is restored. We study this circumstance along the three routes to chaos in low-dimensional nonlinear maps where the attractors at the transitions, between regular and chaotic behavior, drive phase-space contraction for ensembles of trajectories. We illustrate this circumstance for properties of systems that find descriptions in terms of nonlinear maps. These are size-rank functions, urbanization and similar processes, and settings where frequency locking takes place.

[Parametrial involvement in cervical cancer IB1 patients treated with radical hysterectomy in Peru]. / Compromiso parametrial en pacientes con cáncer de cérvix IB-1 tratadas con histerectomía radical en Perú

OBJECTIVES: To assess the frequency and factors associated with parametrial involvement (PI) in patients with cervical cancer IB1 who underwent radical hysterectomy (RH) at the National Institute of Neoplastic Diseases in Peru. MATERIALS AND METHODS: Cross-sectional study of 214 patients with cervical cancer IB1 undergoing RH with pelvic lymphadenectomy between 2007 and 2012. The population was compared with and without clinicopathological variables associated with PI such as age, tumor size, depth of infi histological grade and pelvic lymph node involvement. RESULTS: Ten patients (4.6%) had PI. Multivariate analysis showed that the depth of invasion (OR 8.37, 95% CI 1.24- 56.41, p=0.029) and pelvic node involvement (OR 18.03; 95% CI 3.30-98.44, p=0.001) were all independent predictors of PI. The presence of tumor size = 2 cm, invasion = 10 mm, absence of lymphovascular permeation and absence of nodal involvement identifi 97 patients at low risk of PI, none of whom had PI pathology. CONCLUSIONS: We confi the low frequency of PI in the cervical cancer IB1 subgroup and its association with the depth of invasion and lymph node involvement suggested in previous studies. This information will allow the identifi of a subgroup of patients at low risk of PI for less radical surgery in our institution.