RHOJ as a novel mechanosensitive modulator of endothelial inflammation.

Physiological high shear stress (HSS), a frictional force generated by flowing blood, is essential for endothelial homeostasis under normal physiological conditions. HSS suppresses atherosclerosis by inhibiting endothelial inflammation. However, the molecular mechanisms underlying this process have not been fully elucidated. Here, we report that HSS downregulated the mRNA and protein levels of ras homolog family member J (RHOJ) in endothelial cells (ECs). Silencing endogenous RHOJ expression decreased the mRNA and protein levels of proinflammatory vascular cell adhesion molecule 1 (VCAM-1) and intercellular cell adhesion molecule 1 (ICAM-1) in ECs, leading to a reduction in monocyte adhesion to ECs. Conversely, the overexpression of RHOJ had the opposite effect. RNA-sequencing analysis uncovered several differentially expressed genes (such as yes-associated protein 1 (YAP1),heme oxygenase-1 (HO1), and monocyte chemoattractant protein-1 (MCP1)) and pathways (such as nuclear factor-kappa B (NF-κB), fluid shear stress and atherosclerosis, and cell adhesion pathways) as RHOJ targets. Additionally, HSS was observed to alleviate endothelial inflammation by inhibiting RHOJ expression. Finally, methylated RNA immunoprecipitation sequencing (MeRIP-seq) illustrated that fluid shear stress regulates RHOJ expression in an N6-methyladenosine (m6A)-dependent manner. Mechanistically, the RNA m6A writer, methyltransferase 3 (METTL3), and the RNA m6A readers, YTH N6-methyladenosine RNA-binding protein F 3 (YTHDF3) and YTH N6-methyladenosine RNA-binding protein C 1/2 (YTHDC1/2), are involved in this process. Taken together, our data demonstrate that HSS-induced downregulation of RHOJ contributes to endothelial homeostasis by suppressing endothelial inflammation and that RHOJ inhibition in ECs is a promising therapeutic strategy for endothelial dysfunction.

Short-term effects of fresh mother's own milk in very preterm infants.

Fresh mother's own milk (MOM) can protect preterm infants from many complications. Often MOM is pasteurized for safety, which can deactivate cellular and bioactive components with protective benefits. Questions remain regarding whether pasteurized MOM provides the same benefits as fresh MOM. The aim of this study was to evaluate the association and feasibility of feeding very preterm infants with fresh MOM. This prospective cohort study included 157 very preterm infants born before 32 weeks' gestational age and with a birthweight below 1500 g. Of these, 82 infants were included in the fresh MOM without any processing group and 75 infants were included in the pasteurized never-frozen MOM (PNFMOM) group. The mortality rate, survival rate without severe complication, incidence of complications, feeding indexes and growth velocities were compared to assess the association and feasibility of feeding fresh MOM. Compared with the PNFMOM group, the fresh MOM group had a higher survival rate without severe complications (p = 0.014) and a lower incidence of bronchopulmonary dysplasia (p = 0.010) after adjustment for confounders. The fresh MOM group regained birthweight earlier (p = 0.021), reached total enteral feeding earlier (p = 0.024), and received total parenteral nutrition for less time (p = 0.045). No adverse events associated with fresh MOM feeding were recorded. Feeding fresh MOM may reduce the incidence of complications in very premature infants. Fresh MOM was shown to be a feasible feeding strategy to improve preterm infants' outcomes.

One-step dynamic analysis of growth kinetics of Bacillus cereus from spores in simulated fried rice - Model development, validation, and Marko Chain Monte Carlo simulation.

Bacillus cereus is a spore-forming pathogen capable of producing an emetic toxin and several diarrheal enterotoxins that may cause outbreaks of foodborne illness often associated with rice-based and other farinaceous foods. Therefore, the objective of this study was to investigate the growth kinetics of B. cereus from spores in simulated egg fried rice. The growth of B. cereus was observed under dynamic conditions. Three independent growth curves were analyzed simultaneously using a one-step dynamic analysis (OSDA) to determine the kinetic parameters. The results showed that the minimum, optimum, and maximum growth temperatures were 11.8, 40.8, and 50.6 °C, respectively, with an optimum specific growth rate of 2.4 per h. The root-mean-square-error (RMSE) of model development was 0.4 log CFU/g. Deterministic validation with another 3 independent dynamic temperature profiles showed a RMSE of 0.5 log CFU/g. With Markov Chain Monte Carlo simulation, the RMSE of prediction was only 0.3 log CFU/g. This study proved that OSDA is an effective and efficient method for quickly developing integrated predictive models and estimating kinetic parameters. The resulting integrated model can be used to accurately predict the growth of B. cereus and for managing its risks associated with egg fried rice. The developed kinetic models also can be used to guide restaurant owners and catering establishments to properly prepare and store egg fried rice and other related products to prevent the growth of B. cereus. According to the model, the growth of mesophilic B. cereus is unlikely to occur if the food is stored below 10 °C.

Effect of water activity on inactivation of Listeria monocytogenes using gaseous chlorine dioxide - A kinetic analysis.

The aim of this study was to investigate the effect of water activity (aw) on inactivation of Listeria monocytogenes using gaseous chlorine dioxide (ClO2 (g)) under room temperature. Surface-inoculated tryptic soy agar (TSA) plates adjusted to 9 different water activity levels ranging from 0.994 to 0.429 were used as samples exposed to ClO2 (g) at 150, 250, and 350 ppm for different durations of treatment time. Results showed that the antimicrobial effect of ClO2 (g) significantly decreases as the aw level and ClO2 (g) concentration decrease. Nonlinear models, such as the modified Chick model and the Weibull model, were used to describe the inactivation kinetics of L. monocytogenes. The results showed that the modified Chick model, which is based on chemical reaction kinetics, was more suitable to describe the inactivation of L. monocytogenes (RMSE < 0.5 log CFU/g) than the Weibull model (RMSE < 1.0 log CFU/g). A multiple regression model was developed for the describing the effect of aw and ClO2 (g) concentration on bacterial inactivation. The results of this study may be used to design ClO2 (g) treatment processes to inactivate L. monocytogenes in low-moisture foods.

Dynamic kinetic analysis of growth of Listeria monocytogenes in pasteurized cow milk.

The objective of this study was to develop a dynamic model for predicting the growth of Listeria monocytogenes in pasteurized cow milk under fluctuating temperature conditions during storage and temperature abuse. Six dynamic temperature profiles that simulated random fluctuation patterns were designed to change arbitrarily between 4 and 30°C. The growth data collected from 3 independent temperature profiles were used to determine the kinetic parameters and construct a growth model combining the primary and secondary models using a 1-step dynamic analysis method. The results showed that the estimated minimum growth temperature and maximum cell concentration were 0.6 ± 0.2°C and 7.8 ± 0.1 log cfu/mL (mean ± standard error), with the root mean square error (RMSE) only 0.3 log cfu/mL for model development. The model and the associated kinetic parameters were validated using the data collected under both dynamic and isothermal conditions, which were not used for model development, to verify the accuracy of prediction. The RMSE of prediction was approximately 0.3 log cfu/mL for fluctuating temperature profiles, and it was between 0.2 and 1.1 log cfu/mL under certain isothermal temperatures (2-30°C). The resulting model and kinetic parameters were further validated using 3 growth curves at 4, 7, and 10°C arbitrarily selected from ComBase (www.combase.cc). The RMSE of prediction was 0.8, 0.4, and 0.5 log cfu/mL, respectively, for these curves. The validation results indicated the predictive model was reasonably accurate, with relatively small RMSE. The model was then used to simulate the growth of L. monocytogenes under a variety of continuous and square-wave temperature profiles to demonstrate its potential application. The results of this study showed that the model developed in this study can be used to predict the growth of L. monocytogenes in contaminated milk during storage.

Growth of Clostridium perfringens in cooked chicken during cooling: One-step dynamic inverse analysis, sensitivity analysis, and Markov Chain Monte Carlo simulation.

The objective of this study was to determine the kinetic parameters and apply Markov Chain Monte Carlo (MCMC) simulation to predict the growth of Clostridium perfringens from spores in cooked ground chicken meat during dynamic cooling. Inoculated samples were exposed to various cooling conditions to observe dynamic growth. A combination of 4 cooling profiles was used in one-step inverse analysis with the Baranyi model as the primary model and the cardinal parameters model as the secondary model. Six kinetic parameters of the Baranyi model and the cardinal parameters model, including Q0, Ymax, µopt, Tmin, Topt, and Tmax, were estimated. The estimated Tmin, Topt, and Tmax were 14.8, 42.9, and 50.5 °C, respectively, with a µopt of 5.25 h-1 and maximum cell density of 8.4 log CFU/g. Correlation analysis showed that both Q0 and Ymax are weakly correlated to other parameters, while the remaining parameters are mostly mildly to strongly correlated with each other. Although it may be difficult to estimate highly correlated parameters using a single temperature profile, one-step analysis with multiple different temperature profiles helped estimate them successfully. The estimated parameters were used as the prior information to construct the posterior distribution for Bayesian analysis. MCMC simulation was used to predict the bacterial growth using different dynamic temperature profiles for validation of the accuracy of the predictive models. The MCMC simulation results showed that the Bayesian analysis produced more accurate predictions of bacterial growth during cooling than the deterministic method. With Bayesian analysis, the root-mean-square-error (RMSE) of prediction was only 0.1 log CFU/g with all residual errors within ±0.25 log CFU/g. Therefore, Bayesian analysis is recommended for predicting the growth of C. perfringens in cooked meat during cooling.

Effect of combination of Oxyrase and sodium thioglycolate on growth of Clostridium perfringens from spores under aerobic incubation.

Clostridium perfringens is a strictly anaerobic pathogen that requires absence of oxygen for its growth in laboratory experiments, which is usually attained by using an anaerobic chamber or anaerobic jars. However, it has been demonstrated that C. perfringens may survive for short periods of times due to its adaptive response to O2. Therefore, the objective of this study was to explore the application of Oxyrase (OX) and sodium thioglycolate (ST) as oxygen scavengers, used alone or in combination, for observation of the growth of C. perfringens under aerobic incubation. The growth of C. perfringens from spores in Schaedler Anaerobe Agar containing different levels and combinations of OX and ST was observed at temperatures between 20 and 50 °C. The kinetic parameters, including lag time, specific growth rate, and maximum cell concentrations in the stationary phase, were determined. The results indicated that ST at concentrations of 0.025 and 0.05% (w/w), although allowing eventual growth of C. perfringens, prolonged its lag times, while OX at 1.5% only allowed growth at a lower growth rate in comparison to anaerobic incubation. OX at 3% enhanced the growth of C. perfringens at temperatures between 30 and 50 °C, while higher levels of OX were needed in the medium to support the growth of C. perfringens during storage at 25 °C (>6% OX) and 20 °C (>9% OX), due to the effect of temperature on enzyme activity. No significant difference was found in the kinetic parameters of C. perfringens incubated aerobically with OX and the control (without OX or ST) in an anaerobic chamber. Therefore, OX at appropriate concentrations may allow the observation of the growth of C. perfringens under aerobic incubation conditions without the need of an anaerobic device.

Effect of temperature on the growth of Staphylococcus aureus in ready-to-eat cooked rice with pork floss.

Cooked rice with pork floss (CRPF) wrapped in dried seaweed is one of the most popular ready-to-eat (RTE) foods in many Asian countries, particularly in Taiwan. The products are susceptible to Staphylococcus aureus contamination and temperature abuse during manufacturing, distribution, and storage. The objective of this study was to examine the effect of temperature on its growth in RTE CRPF for use in risk assessment and prevention of staphylococcal food poisoning (SFP). Inoculated CRPF samples were stored at 4, 12, 18, 25, and 35°C, and the change in the populations of S. aureus during storage were analyzed using three primary models to determine specific growth rate (µmax), lag-phase duration (λ), and maximum population density (ymax). The Ratkowsky square-root and Huang square-root (HSR) models were used as the secondary models to describe the effect of temperature on µmax, and a linear and an exponential regression models were used to describe the effect of temperature on λ and ymax, respectively. The model performance was evaluated by the root mean square error (RMSE), bias factor (Bf), and accuracy factor (Af) when appropriate. Results showed that three primary models were suitable for describing the growth curves, with RMSE ≤ 0.3 (log MPN/g). Using µmax obtained from the Huang model, the minimum growth temperature (Tmin) estimated by the HSR model was 7.0°C, well in agreement with the reported Tmin. The combination of primary and secondary models for predicting S. aureus growth was validated by additional growth curves at 30°C, which showed that the RMSE was 0.6 (log MPN/g). Therefore, the developed models were acceptable for predicting the growth of S. aureus in CRPF under likely temperature abuse conditions and can be applied to assess the risk of S. aureus in CRPF and design temperature controls to reduce the risk of SFP.

Growth kinetics of Staphylococcus aureus and background microorganisms in camel milk.

Staphylococcus aureus is a common foodborne pathogen that is ubiquitous in nature. Consumption of contaminated foods, such as dairy products, can lead to food poisoning caused by heat-stable staphylococcal toxins that are not easily destroyed during pasteurization. The objective of this study was to investigate the growth kinetics of S. aureus and background microorganisms in camel milk stored at different temperatures between 8 and 43°C using one-step kinetic analysis to estimate the kinetic parameters from the observed growth curves. The growth of S. aureus showed apparent lag, exponential, and stationary phases, whereas no or negligible lag phase was observed for background microorganisms. Data analysis showed that the estimated minimum, optimum, and maximum growth temperatures were 5.9, 42.0, and 49.2°C for S. aureus, and 3.0, 38.6, and 49.2°C for the background microorganisms, respectively. The estimated optimum specific growth rate was higher for S. aureus (1.24 h-1) than for background microorganisms (0.995 h-1). This study found that camel milk may inhibit the growth of S. aureus, as it exhibits a lower specific growth rate than that in cow milk or cooked potato. It also has a longer lag phase than that in cow milk at comparable temperature ranges. This unique property is probably related to the presence of some antimicrobial compounds naturally occurring in camel milk. Validation of kinetic parameters and models showed that the root mean square error of prediction was only 0.5 log cfu/mL for S. aureus and background microorganisms, suggesting that the models are reasonably accurate. These models can be used for conducting risk assessments of S. aureus and predicting the general microbiological shelf life of camel milk to prevent foodborne staphylococcal poisoning.

Effect of high pressure treatment on the survival of Shiga toxin-producing Escherichia coli in strawberry puree.

Most fresh produce, such as strawberries, receives minimal processing and is often eaten raw. Contamination of produce with pathogenic bacteria may occur during growth, harvest, processing, transportation, and storage (abuse temperature) and presents a serious public health risk. Strawberries have been implicated in an outbreak of Escherichia coli O157:H7 infection that sickened 15 people, including one death. Strawberries may also be contaminated by other serogroups of non-O157 Shiga toxin-producing E. coli (STEC), including O26, O45, O103, O111, O121 and O145, which have become known as the "Big Six" or "Top Six" non-O157 STECs. The objective of this research was to explore the potential application of high pressure processing (HPP) treatment to reduce or eliminate STECs in fresh strawberry puree (FSP). FSP, inoculated with a six-strain cocktail of the "Big Six" non-O157 STEC strains or a five-strain cocktail of E. coli O157:H7 in vacuum-sealed packages, were pressure-treated at 150, 250, 350, 450, 550, and 650 MPa (1 MPa = 10(6) N/m(2)) for 5, 15, and 30 min. HPP treatment, at 350 MPa for ≥5 min, significantly reduced STECs in FSP by about 6-log CFU/g from the initial cell population of ca. 8-log CFU/g. Cell rupture, observed by scanning electron microscopy (SEM), demonstrated that the HPP treatments can be potentially used to control both non-O157 and O157:H7 STECs in heat sensitive products.

Growth and no-growth boundary of Clostridium perfringens in cooked cured meats - A logistic analysis and development of critical control surfaces using a solid growth medium.

This study was conducted to evaluate the effect of sodium nitrite (NaNO2, 100-200 ppm), sodium erythorbate (SE, 0-547 ppm), sodium tripolyphosphate (STPP, 0-0.5 %), and sodium chloride (NaCl, 2-3 %) on growth of C. perfringens using a solid growth medium and to develop a growth/no-growth boundary (critical control surface, or CCS) to prevent its growth in cooked cured meat under the optimal temperature condition. Melted Shahidi Ferguson Perfringens (SFP) agar, inoculated with a 3-strain spore cocktail and mixed with NaNO2, SE, STPP, and NaCl according to a Box-Behnken response surface experimental design, was dispersed in 96-well microplates and incubated anaerobically in an incubator programmed to remain at 4 °C for 24 h, heat to 80 °C in 1.75 h, quickly (0.5 h) cool to 46 °C (optimum temperature), and then maintain at 46 °C overnight. The plates were examined optically and visually for colony formation. Any well free of growth was designated as no-growth. Logistic regression was used to analyze the growth probability (P) as affected by NaNO2, SE, STPP, and NaCl and define a CSS as meeting the criterion of P < 1/96. The results showed that STPP and the interactions of SE with NaNO2 and NaCl could reduce the growth probability of C. perfringens in SFP agar. The validation of CCS with ground beef showed an accuracy of 96.3 % for no growth of C. perfringens in the inoculated samples. The results of this study proved that cured meat can be formulated with proper combinations of NaNO2, SE, STPP, and NaCl to prevent the growth of C. perfringens even under the optimum temperature condition, thus preventing food poisoning caused by the growth of this microorganism.

Thermal resistance of Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella in animal fat - Kinetic analysis and mathematical modeling.

The objective of this study was to evaluate the effect of fat on thermal resistance of L. monocytogenes, E. coli O157:H7, and Salmonella spp. A 4-strain cocktail of each microorganism was inoculated to beef tallow and heated isothermally at temperatures between 55 and 80℃. All survival curves did not follow the 1st-order inactivation kinetics but conformed to a two-stage linear pattern. The first stage was markedly less heat-resistant than the second, as manifested by significantly lower D values. The z values of E. coli O157 H7 and Salmonella spp. were 11.8 °C and 12.3 °C in the first stage (z1) but increased to 23.7 °C and 20.8 °C in the second stage (z2), respectively. For L. monocytogenes, while the z values were similar for both stages (z1 = 19.6 °C and z2 = 18.5 °C), the second stage D values are 3.6-5.9 times of those in the first stage. One-step analysis was used to fit the nonlinear curves to the Weibull model, yielding < 1 exponents for the model (0.495, 0.362, and 0.282, respectively, for L. monocytogenes, E. coli O157:H7, and Salmonella spp.), suggesting gradually increased thermal resistance during heating. The experimental results showed that these microorganisms could resist heating for longer time and at higher temperatures in tallow than they do in regular meats containing lower levels of fat. The kinetic models can be used to develop thermal processes to properly inactivate pathogens contaminated in the fat portions of meat products or other high fat products.

One-step analysis of growth kinetics of mesophilic Bacillus cereus in liquid egg yolk during treatment with phospholipase A2: Model development and validation.

Liquid egg yolk (LEY) is often treated with phospholipase A2 (PLA2) to improve its emulsifying capacity and thermal stability. However, this process may allow certain pathogens to grow. The objective of this study was to evaluate the growth kinetics of mesophilic Bacillus cereus in LEY during PLA2 treatment. Samples, inoculated with B. cereus vegetative cells, were incubated isothermally at different temperatures between 9 and 50 °C to observe the bacterial growth and survival. Under the observation conditions, bacterial growth occurred between 15 and 48 °C, but not at 9 and 50 °C. The growth curves were analyzed using the USDA IPMP-Global Fit, with the no-lag phase model as the primary model in combination with either the cardinal temperatures model (CTM) or the Huang square-root model (HSRM) as the secondary model. While similar maximum growth temperatures (Tmax) were determined (48.4 °C for HSRM and 48.1 °C for CTM), the minimum growth temperature (Tmin) of the HSRM more accurately described the lower limit (9.26 °C), in contrast to 6.51 °C for CTM, suggesting that the combination of the no-lag phase model and HSRM was more suitable to describe the growth of mesophilic B. cereus in LEY. The root mean square error (RMSE) of model validation and development was <0.5 log CFU/g, indicating the combination of the no-lag phase model and HSRM could predict the growth of mesophilic B. cereus in LEY during PLA2 treatment. The results of this study may allow the food industry to choose a suitable temperature for PLA2 treatment of LEY to prevent the growth of mesophilic B. cereus.

Numerical simulation of heat transfer during meat ball cooking and microbial food safety enhancement.

This study was conducted to apply the finite volume method (FVM) to solve the partial differential equation (PDE) governing the heat transfer process during meat cooking with convective surface conditions. For a one-dimensional, round-shaped food, such as meat balls, the domain may be divided into shells of equal thickness, with energy balance established for each adjacent shell using in the finite difference scheme (FDS) to construct a set of finite difference equations, which were then solved simultaneously using the FORTRAN language and the IVPAG subroutine of the International Mathematics and Statistics Library. The FDS is flexible for temperature-dependent physical properties of foods, such as thermal conductivity (k), specific heat (Cp ), thermal diffusivity (α), and boundary conditions, for example, surface heat transfer coefficient (h), to predict the dynamic temperature profiles in beef and chicken meat balls cooked in an oven. Once the FVM model was established and validated, it was used to simulate the dynamic temperature profiles during cooking, which were then used in combination with the general method to evaluate the thermal lethality of Shiga toxin-producing Escherichia coli and Salmonella spp. using D and z values in ground meats during cooking. The method can be applied to design cooking processes that effectively inactivate foodborne pathogens while maintaining the quality of cooked meats and evaluate the adequacy of a cooking process. PRACTICAL APPLICATION: The temperature dependences of thermal conductivity (k) and thermal diffusivity (α) of raw ground beef and ground chicken meats were measured. These thermal properties were then used in numerical simulation to predict the dynamic heating temperature profile and thermal lethality of ground beef and chicken meat balls. The numerical simulation method may be used to optimize and evaluate thermal processes and ensure the inactivation of pathogens in meat products during cooking.

Decadal Changes in Population Structures of Rare Oak Species Quercus chungii.

Quercus chungii, a rare and endangered endemic tree species, is found exclusively in subtropical regions of China. Understanding the population structure and temporal dynamics of Q. chungii is pivotal for effective conservation and restoration of its populations and associated ecosystems. However, large knowledge gaps remain about its population structure and temporal change and its key demographic rates across size classes. In this study, we investigated the population structures of Q. chungii in 2013 and 2023 in a nature reserve specifically established to better conserve this species and its associated ecosystems. We found that Q. chungii increased in its overall abundance and tree size in the past decade, suggesting active regeneration and a rapid growth rate for this species and the effectiveness of past conservation efforts. The age structure in 2023 showed a pyramid shape, with a sharp decline in the numbers of individuals from germinated seeds to seedlings and from seedlings to saplings. These led to the low numbers of seedlings and saplings and high age-specific death probabilities at the early developmental stages. These results indicated potential risks of future population decline. These risks may have already manifested over the past decade, as a high mortality rate during the seedling-to-sapling transition could be one of the primary reasons contributing to the decreased proportion of saplings in 2023 compared to 2013. We propose that future studies may benefit from in-depth studies on the regeneration processes of Q. chungii by considering seed predation and germination under changing climate. This study improves the prediction of population development of Q. chungii, thereby offering theoretical guidance essential for its conservation.

Whole-genome sequencing and characterization of Escherichia coli human isolate DP033.

The whole-genome sequence of Escherichia coli strain DP033 is reported here. DP033 was isolated from a human rectal specimen in Tilburg, the Netherlands. In silico analysis showed that DP033 possessed 36 virulence-related genes and is a presumptive extraintestinal pathogenic E. coli and uropathogenic E. coli strain.

Gene expression profiling of a nisin-sensitive Listeria monocytogenes Scott A ctsR deletion mutant.

Listeria monocytogenes is a food-borne pathogen of significant threat to public health. Nisin is the only bacteriocin that can be used as a food preservative. Due to its antimicrobial activity, it can be used to control L. monocytogenes in food; however, the antimicrobial mechanism of nisin activity against L. monocytogenes is not fully understood. The CtsR (class III stress gene repressor) protein negatively regulates the expression of class III heat shock genes. A spontaneous pressure-tolerant ctsR deletion mutant that showed increased sensitivity to nisin has been identified. Microarray technology was used to monitor the gene expression profiles of the ctsR mutant under treatments with nisin. Compared to the nisin-treated wild type, 113 genes were up-regulated (>2-fold increase) in the ctsR deletion mutant whereas four genes were down-regulated (<-2-fold decrease). The up-regulated genes included genes that encode for ribosomal proteins, membrane proteins, cold-shock domain proteins, translation initiation and elongation factors, cell division, an ATP-dependent ClpC protease, a putative accessory gene regulator protein D, transport and binding proteins, a beta-glucoside-specific phosphotransferase system IIABC component, as well as hypothetical proteins. The down-regulated genes consisted of genes that encode for virulence, a transcriptional regulator, a stress protein, and a hypothetical protein. The gene expression changes determined by microarray assays were confirmed by quantitative real-time PCR analyses. Moreover, an in-frame deletion mutant for one of the induced genes (LMOf2365_1877) was constructed in the wild-type L. monocytogenes F2365 background. ΔLMOf2365_1877 had increased nisin sensitivity compared to the wild-type strain. This study enhances our understanding of how nisin interacts with the ctsR gene product in L. monocytogenes and may contribute to the understanding of the antibacterial mechanisms of nisin.

Growth kinetics of Listeria monocytogenes and spoilage microorganisms in fresh-cut cantaloupe.

The main objective of this study was to investigate the growth kinetics of Listeria monocytogenes and background microorganisms in fresh-cut cantaloupe. Fresh-cut cantaloupe samples, inoculated with three main serotypes (1/2a, 1/2b, and 4b) of L. monocytogenes, were incubated at different temperatures, ranging from 4 to 43 °C, to develop kinetic growth models. During storage studies, the population of both background microorganisms and L. monocytogenes began to increase almost immediately, with little or no lag phase for most growth curves. All growth curves, except for two growth curves of L. monocytogenes 1/2a at 4 °C, developed to full curves (containing exponential and stationary phases), and can be described by a 3-parameter logistic model. There was no significant difference (P = 0.28) in the growth behaviors and the specific growth rates of three different serotypes of L. monocytogenes inoculated to fresh-cut cantaloupe. The effect of temperature on the growth of L. monocytogenes and spoilage microorganisms was evaluated using three secondary models. For L. monocytogenes, the minimum and maximum growth temperatures were estimated by both the Ratkowsky square-root and Cardinal parameter models, and the optimum temperature and the optimum specific growth rate by the Cardinal parameter model. An Arrhenius-type model provided more accurate estimation of the specific growth rate of L. monocytogenes at temperatures <4 °C. The kinetic models developed in this study can be used by regulatory agencies and food processors for conducting risk assessment of L. monocytogenes in fresh-cut cantaloupe, and for estimating the shelf-life of fresh-cut products.

Shelf-life boundaries of Listeria monocytogenes in cold smoked salmon during refrigerated storage and temperature abuse.

Cold smoked salmon (CSS) is a high-value ready-to-eat product, but it generally has a short shelf-life even under refrigeration and can support the growth of Listeria monocytogenes. Therefore, the objective of this study was to examine the growth and survival of L. monocytogenes in CSS during refrigerated storage and temperature abuse. The growth and survival data of L. monocytogenes (116 records, 465 data points) were retrieved from ComBase (https://www.combase.cc). All records contained storage time and temperature, but other information (aw, pH, and salt) was not fully documented. Each data point, normalized with the initial population to calculate relative growth (RG, log CFU/g), was used to classify the probability of growth. Eighty percent (80%) of the data were randomly sampled for examining the effect of storage time and temperature on growth of L. monocytogenes, while the remaining 20% were set aside for model validation. Logistic regression was used to develop a model for classifying L. monocytogenes growth according to 7 different control thresholds (CT), ranging from 0 to 3 log CFU/g in RG. A probability threshold was set to judge if the bacterial growth has exceeded a CT. The validation showed > 89% of true negative rate for not exceeding the control thresholds. A dynamic method was then developed and demonstrated to predict the growth probabilities under fluctuating temperature conditions. The result of this study suggested that storage time and temperature could be used to predict the growth of L. monocytogenes in CSS and to control listeriosis using a risk-based strategy. It can be used by the retailers and consumers to determine if a packaged product is safe to consume based on its time and temperature history.

Enhanced external counterpulsation improves dysfunction of forearm muscle caused by radial artery occlusion.

Objective: This study aimed to investigate the therapeutic effect of enhanced external counterpulsation (EECP) on radial artery occlusion (RAO) through the oscillatory shear (OS) and pulsatile shear (PS) models of human umbilical vein endothelial cells (HUVECs) and RAO dog models. Methods: We used high-throughput sequencing data GSE92506 in GEO database to conduct time-series analysis of functional molecules on OS intervened HUVECs, and then compared the different molecules and their functions between PS and OS. Additionally, we studied the effect of EECP on the radial artery hemodynamics in Labrador dogs through multi-channel physiological monitor. Finally, we studied the therapeutic effect of EECP on RAO at the histological level through Hematoxylin-Eosin staining, Masson staining, ATPase staining and immunofluorescence in nine Labrador dogs. Results: With the extension of OS intervention, the cell cycle decreased, blood vessel endothelial cell proliferation and angiogenesis responses of HUVECs were down-regulated. By contrast, the inflammation and oxidative stress responses and the related pathways of anaerobic metabolism of HUVECs were up-regulated. Additionally, we found that compared with OS, PS can significantly up-regulate muscle synthesis, angiogenesis, and NO production related molecules. Meanwhile, PS can significantly down-regulate inflammation and oxidative stress related molecules. The invasive arterial pressure monitoring showed that 30Kpa EECP treatment could significantly increase the radial artery peak pressure (p = 0.030, 95%CI, 7.236-82.524). Masson staining showed that RAO significantly increased muscle interstitial fibrosis (p = 0.002, 95%CI, 0.748-2.128), and EECP treatment can reduce this change (p = 0.011, 95%CI, -1.676 to -0.296). ATPase staining showed that RAO significantly increased the area of type II muscle fibers (p = 0.004, 95%CI, 7.181-25.326), and EECP treatment could reduce this change (p = 0.001, 95%CI, -29.213 to -11.069). In addition, immunofluorescence showed that EECP increased angiogenesis in muscle tissue (p = 0.035, 95%CI, 0.024-0.528). Conclusion: EECP improves interstitial fibrosis and hypoxia, and increases angiogenesis of muscle tissue around radial artery induced by RAO.