Dataset Information

Analyzing Thermal Stability of Cell Membrane of Salmonella Using Time-Multiplexed Impedance Sensing.

ABSTRACT: Heat treatment is one of the most widely used methods for inactivation of bacteria in food products. Heat-induced loss of bacterial viability has been variously attributed to protein denaturation, oxidative stress, or membrane leakage; indeed, it is likely to involve a combination of these processes. We examine the effect of mild heat stress (50-55°C for ?12 min) on cell permeability by directly measuring the electrical conductance of samples of Salmonella enterica serovar Typhimurium to answer a fundamental biophysical question, namely, how bacteria die under mild heat stress. Our results show that when exposed to heat shock, the cell membrane is damaged and cells die mainly due to the leakage of small cytoplasmic species to the surrounding media without lysis (confirmed by fluorescent imaging). We measured the conductance change, ?Y, of wild-type versus genetically modified heat-resistant (HR) cells in response to pulse and ramp heating profiles with different thermal time constants. In addition, we developed a phenomenological model to correlate the membrane damage, cytoplasmic leakage, and cell viability. This model traces the differential viability and ?Y of wild-type and HR cells to the difference in the effective activation energies needed to permeabilize the cells, implying that HR cells are characterized by stronger lateral interactions between molecules, such as lipids, in their cell envelope.

SUBMITTER: Ebrahimi A

PROVIDER: S-EPMC5985002 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature

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Analyzing Thermal Stability of Cell Membrane of Salmonella Using Time-Multiplexed Impedance Sensing.

Ebrahimi Aida A Csonka Laszlo N LN Alam Muhammad A MA

Biophysical journal 20180201 3

Heat treatment is one of the most widely used methods for inactivation of bacteria in food products. Heat-induced loss of bacterial viability has been variously attributed to protein denaturation, oxidative stress, or membrane leakage; indeed, it is likely to involve a combination of these processes. We examine the effect of mild heat stress (50-55°C for ≤12 min) on cell permeability by directly measuring the electrical conductance of samples of Salmonella enterica serovar Typhimurium to answer ...[more]

PMID: 29414707

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Project description:Analysis of transcriptome in moss Physcomitrella patens CNGCb null mutant at 25 and 34 degrees C for 30 minutes. Results provide insight into role of CNGCb in acquired thermotolerance induced by non-lethal heat treatment. Typically at dawn of a hot summer day, land plants need precise molecular thermometers to sense harmless increments in the ambient temperature to timely develop a heat-shock response (HSR) and accumulate protective heat shock proteins (Hsps), in anticipation of upcoming harmful temperatures at mid-day. Here, we found that the CNGCb gene from Physcomitrella patens and its Arabidopsis ortholog CNGC2, encode for a component of cyclic nucleotide gated Ca2+ channels acting as the primary thermosensors of land plant cells. Disruption of CNGCb or CNGC2 produced a hyper-thermosensitive phenotype, giving rise to a HSR and acquired thermotolerance at significantly milder heat-priming treatments than in wild type plants. In an aequorin-expressing moss, CNGCb loss-of-function caused altered Ca2+ signaling and a sustained Ca2+ influx. Patch clamp recordings on moss protoplasts showed the presence of three distinct thermo-responsive Ca2+-channels in wild type cells. Deletion of CNGCb led to a total absence of one, and it increased the open probability of the remaining two thermo-responsive Ca2+ channels. Thus, both in Arabidopsis and moss, CNGC2 and CNGCb are expected to form with other related CNGCs, heteromeric Ca2+ channels in the plasma membrane that respond to mild increments in the ambient temperature by triggering an optimal HSR, leading to the onset of plant acquired thermotolerance. The WT moss tissues were heat-shocked for a half an hour at 34°C and 38°C and CNGCb at 25and 34°C followed by liquid nitrogen freezing. Total RNA was isolated using RNeasy Mini Kit (QIAGEN, Hilden, Germany) and two biological replicate samples for each treatment, were extracted. An Agilent-certified microarray service lab (MOgene, LC, St. Louis, MO, USA) was used to verify the integrity of the RNA and perform the microarray experiments. Two biological replicates were performed.

Project description:Uncertainty analysis is the process of identifying limitations in scientific knowledge and evaluating their implications for scientific conclusions. In the context of microbial risk assessment, the uncertainty in the predicted microbial behavior can be an important component of the overall uncertainty. Conventional deterministic modeling approaches which provide point estimates of the pathogen's levels cannot quantify the uncertainty around the predictions. The objective of this study was to use Bayesian statistical modeling for describing uncertainty in predicted microbial thermal inactivation of Salmonella enterica Typhimurium DT104. A set of thermal inactivation data in broth with water activity adjusted to 0.75 at 9 different temperature conditions obtained from the ComBase database (www.combase.cc) was used. A log-linear microbial inactivation was used as a primary model while for secondary modeling, a linear relation between the logarithm of inactivation rate and temperature was assumed. For comparison, data were fitted with a two-step and a global Bayesian regression. Posterior distributions of model's parameters were used to predict Salmonella thermal inactivation. The combination of the joint posterior distributions of model's parameters allowed the prediction of cell density over time, total reduction time and inactivation rate as probability distributions at different time and temperature conditions. For example, for the time required to eliminate a Salmonella population of about 107 CFU/ml at 65°C, the model predicted a time distribution with a median of 0.40 min and 5th and 95th percentiles of 0.24 and 0.60 min, respectively. The validation of the model showed that it can describe successfully uncertainty in predicted thermal inactivation with most observed data being within the 95% prediction intervals of the model. The global regression approach resulted in less uncertain predictions compared to the two-step regression. The developed model could be used to quantify uncertainty in thermal inactivation in risk-based processing design as well as in risk assessment studies.

Project description:Study objectivesReal-time monitoring of upper airway collapse during sleep could be instrumental for studies in biomechanics of obstructive sleep apnea (OSA) and selecting individualized treatment modalities. Although some imaging techniques are used under sedated sleep, none are available during the entire natural sleep process. We hypothesized that electrical impedance tomography (EIT) can be used for noninvasive continuous imaging of the upper airway during natural sleep and quantifying upper airway collapse in terms of its size.MethodsAfter determining surface landmarks to attach the electrodes for monitoring the retroglossal airway, EIT was conducted in 10 healthy participants. As a feasibility test of EIT in detecting upper airway collapse, transient airway closure was induced by the swallowing maneuver. These EIT images were confirmed by simultaneous magnetic resonance imaging (MRI) scans. Subsequently, EIT scans were conducted in 7 healthy participants and 10 patients with OSA under nonsedated sleep to determine whether it could identify upper airway narrowing or collapse. Respiratory events were identified by concurrent polysomnography (PSG).ResultsSwallowing-induced airway closure was identified successfully in all 10 participants on simultaneous EIT and MRI scans. Sizes and positions of the upper airway closures in reconstructed EIT images were well correlated with those in magnetic resonance images. Obstructive hypopnea and apnea were detected successfully by EIT in 10 patients with OSA, and no significant changes in EIT data were observed in 7 healthy participants during concurrent EIT and PSG tests. Additionally, conductivity changes in the airway were greater during obstructive apnea than during hypopnea (64.3% versus 26.3%, respectively; P < .001) compared with those during baseline respiration.ConclusionsEIT could be a useful real-time monitoring device for detecting upper airway narrowing or collapse during natural sleep in patients with OSA. Currently, changes in the upper airway size can be estimated with good accuracy, but shape estimation needs future improvements in the EIT image quality.

Dataset Information

Analyzing Thermal Stability of Cell Membrane of Salmonella Using Time-Multiplexed Impedance Sensing.

Publications

Analyzing Thermal Stability of Cell Membrane of Salmonella Using Time-Multiplexed Impedance Sensing.

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