Fabrication and characterization of dual-functional porous starch with both emulsification and antioxidant properties.

Starchy materials with good antioxidant, emulsification and adsorption properties have potential applications in industry. To improve these properties, a Dual-functional porous starch was prepared through one-pot synthesis. In this case, octenyl succinic anhydride (OSA) and syringic acid (SA) were selected to modify the porous starch (PS) by esterification, with subsequent signals recorded by 1H NMR at 1.2 ppm and FT-IR at 1743 cm-1, indicating the formation of Dual-functional porous starch grafted by OSA and SA. N2 adsorption analysis further proved that the porous structure (2.9 m2g-1) was still maintained after modification. This was followed by measurements of droplet size distribution (34.18 ± 3.80 µm), zeta potential (-39.62 ± 1.89 mV) and emulsion index (85.10 ± 1.76 %), all of which indicated good emulsifying capacity. Meanwhile, results of radical scavenging assay proved that the Dual-functional porous starch had considerable antioxidant properties due to the introduction of SA groups. Besides, the Dual-functional porous starch also showed good resistance to digestion. These findings not only provide a novel strategy for constructing multi-functionalized starchy materials, but also open up potential applications of starch in the food and pharmaceutical industries.

AQRNA-seq for Quantifying Small RNAs.

AQRNA-seq provides a direct linear relationship between sequencing read counts and small RNA copy numbers in a biological sample, thus enabling accurate quantification of the pool of small RNAs. The AQRNA-seq library preparation procedure described here involves the use of custom-designed sequencing linkers and a step for reducing methylation RNA modifications that block reverse transcription processivity, which results in an increased yield of full-length cDNAs. In addition, a detailed implementation of the accompanying bioinformatics pipeline is presented. This demonstration of AQRNA-seq was conducted through a quantitative analysis of the 45 tRNAs in Mycobacterium bovis BCG harvested on 5 selected days across a 20-day time course of nutrient deprivation and 6 days of resuscitation. Ongoing efforts to improve the efficiency and rigor of AQRNA-seq will also be discussed here. This includes exploring methods to obviate gel purification for mitigating primer dimer issues after PCR amplification and to increase the proportion of full-length reads to enable more accurate read mapping. Future enhancements to AQRNA-seq will be focused on facilitating automation and high-throughput implementation of this technology for quantifying all small RNA species in cell and tissue samples from diverse organisms.

Probing covalent and non-covalent interactions between vanillic acid and starch and their effects on digestibility by solid-state NMR.

Intermolecular interactions play a significant role on the physicochemical properties and digestibility of starchy foods. This study investigated the covalent and non-covalent interactions between vanillic acid (VA) and porous starch (PS) as well as their effects on digestibility using solid-state NMR. VA-PS conjugates and mixtures were synthesized and characterized using 1H NMR, FT-IR, SEM and XRD. 13C NMR peaks at 163 ppm and FT-IR signals at 1737 cm-1 indicated the formation of ester bond in VA-PS conjugates. While differences between covalent and non-covalent interactions were also probed by solid-state NMR. The specific binding sites between VA and PS were subsequently identified by 1H13C HETCOR spectra before assessing the impact of covalent and non-covalent interactions on digestibility through an in vitro digestion test. The results revealed 13C chemical shifts of about 2.0 ppm, indicating stronger intermolecular interactions, and reduced mobility of the VA-PS conjugate due to its covalent bonding. Overall, the results showed that the VA-PS conjugate, characterized by stronger covalent interactions, exhibited superior effects in inhibiting starch digestibility compared with non-covalent interactions.

Insight into structure-activity relationships of hydroxycinnamic acids modified porous starch: The effect of phenolic hydroxy groups.

Antioxidant capacity of hydroxycinnamic acids-modified starch mainly depends on their chemical structure. Herein, cinnamic acid as well as meta-substituted and para-substituted cinnamic acid were selected for esterification with porous starch (labelled as CA@PS, m-CA@PS and p-CA@PS), with the successful formation of porous starch (labelled as PS) esters then confirmed by 1H NMR, 13C solid-state NMR and FT-IR spectroscopy. Three PS esters with almost same degrees of substitution (DS) were obtained, and antioxidant assays, including DPPH radical scavenging, reducing power and hydroxyl radical scavenging tests, were subsequently used to evaluate the antioxidant activity of the esterified PS. Overall, CA@PS showed weak antioxidant activity because of the absence of phenolic hydroxy, while p-CA@PS displayed better antioxidant capacity. Because its conjugated structure offered the stronger electron-donating effect, that could enhance antioxidant capacity. Therefore, antioxidant capacity depended significantly on overall chemical structure, including numbers and substitution positions of phenolic hydroxy groups.

Molecular interactions between apigenin and starch with different amylose/amylopectin ratios revealed by X-ray diffraction, FT-IR and solid-state NMR.

Starch can readily form complexes with polyphenols. However, its two components, namely amylose and amylopectin, differ significantly in their ability to complex with phenolic compounds. Given that the mechanism of their interaction is still poorly studied, this work investigated intermolecular interactions between apigenin and starch with different amylose/amylopectin ratios using 1H NMR, FT-IR, XRD, DSC and solid-state NMR. Results showed that corn starch with high amylose/amylopectin ratios had a better complexing ability and higher complexing index with apigenin than amylopectin. Besides, solid-state NMR suggested that the molecular mechanism behind the strong intermolecular interactions between corn starch and apigenin involved hydrogen bonds. Furthermore, the detailed binding sites of hydrogen bonds, that linked by hydroxyl-starch and phenyl-apigenin were also confirmed by 1H13C heteronuclear correlation (HETCOR) spectra. This study revealed the molecular mechanism on amylose/amylopectin complexing with apigenin and provides a theoretical basis for further developing polyphenols in starchy food.

Probing molecular interactions of amylose-morin complex and their effect on antioxidant capacity by 2D solid-state NMR spectroscopy.

Interaction of polyphenols and starch significantly governed the further applications on polyphenol-starchy foods. Elucidation of inter-molecular interaction is, however, a challenge because conventional characterizations could not detect the change of micro-environment caused by weak interactions. Herein, a facile strategy for molecular detection of amylose-polyphenol interactions was reported using two-dimensional solid-state NMR spectroscopy. Amylose-morin complex was prepared and characterized using 1H NMR, FT-IR, DSC, XRD and SEM. Significantly, variation of chemical shifts, splitted peaks and peak width, monitored by 13C CP/MAS and 1H NMR spectra, identified the strong inter-molecular interaction and binding sites. Furthermore, correlated signals from 1H-13C HETCOR confirmed the binding sites of interactions. These findings confirmed the interaction was inter-molecular hydrogen bonds, which generated between hydroxy-3,5,7 of morin and hydroxy groups of amylose. Besides, DPPH radical scavenging and reducing power assay indicated inter-molecular hydrogen bonds are not strong enough to interfere antioxidant capacity of morin.

Anatomical Evidence for Parasympathetic Innervation of the Renal Vasculature and Pelvis.

BACKGROUND: The kidneys critically contribute to body homeostasis under the control of the autonomic nerves, which enter the kidney along the renal vasculature. Although the renal sympathetic and sensory nerves have long been confirmed, no significant anatomic evidence exists for renal parasympathetic innervation. METHODS: We identified cholinergic nerve varicosities associated with the renal vasculature and pelvis using various anatomic research methods, including a genetically modified mouse model and immunostaining. Single-cell RNA sequencing (scRNA-Seq) was used to analyze the expression of AChRs in the renal artery and its segmental branches. To assess the origins of parasympathetic projecting nerves of the kidney, we performed retrograde tracing using recombinant adeno-associated virus (AAV) and pseudorabies virus (PRV), followed by imaging of whole brains, spinal cords, and ganglia. RESULTS: We found that cholinergic axons supply the main renal artery, segmental renal artery, and renal pelvis. On the renal artery, the newly discovered cholinergic nerve fibers are separated not only from the sympathetic nerves but also from the sensory nerves. We also found cholinergic ganglion cells within the renal nerve plexus. Moreover, the scRNA-Seq analysis suggested that acetylcholine receptors (AChRs) are expressed in the renal artery and its segmental branches. In addition, retrograde tracing suggested vagus afferents conduct the renal sensory pathway to the nucleus of the solitary tract (NTS), and vagus efferents project to the kidney. CONCLUSIONS: Cholinergic nerves supply renal vasculature and renal pelvis, and a vagal brain-kidney axis is involved in renal innervation.

The efficacy of nisin against Listeria monocytogenes on cold-smoked salmon at natural contamination levels is concentration-dependent and varies by serotype.

Cold-smoked salmon is a ready-to-eat food product capable of supporting Listeria monocytogenes growth at refrigeration temperatures. While the FDA-approved antimicrobial nisin can be used to mitigate L. monocytogenes contamination, stresses associated with cold-smoked salmon and the associated processing environments may reduce nisin efficacy. A previous study in our laboratory showed that, at high inoculation levels, pre-exposure of L. monocytogenes to sublethal concentrations of quaternary ammonium compounds had an overall detrimental effect on nisin efficacy. The objective of this study was to investigate the impact of nisin concentration and storage temperature on nisin efficacy against L. monocytogenes inoculated on salmon at natural contamination levels. Three L. monocytogenes strains were pre-grown in the presence of sublethal levels of benzalkonium chloride prior to inoculation at ~102 CFU/g on salmon slices that were pre-treated with either 0, 25, or 250 ppm nisin, followed by vacuum-packing and incubation at 4 or 7°C for up to 30 days. L. monocytogenes was enumerated on days 1, 15, and 30 using direct plating and/or most probable number methods. A hurdle model was constructed to describe the odds of complete elimination of L. monocytogenes on salmon and the level of L. monocytogenes when complete elimination was not achieved. Our data showed that (i) nisin efficacy (defined as L. monocytogenes reduction relative to the untreated control) was concentration-dependent with increased efficacy at 250 ppm nisin, and that (ii) 250 ppm nisin treatments led to a reduction in L. monocytogenes prevalence, independent of storage temperature and serotype; this effect of nisin could only be identified since low inoculation levels were used. While lower storage temperatures (i.e., 4°C) yielded lowered absolute L. monocytogenes counts on days 15 and 30 (as compared to 7°C), nisin efficacy did not differ between these two temperatures. Finally, the serotype 1/2b strain was found to be more susceptible to nisin compared with serotype 1/2a and 4b strains on samples incubated at 7°C or treated with 25 ppm nisin. This variation of nisin susceptibility across serotypes, which is affected by both the storage temperature and nisin concentration, needs to be considered while evaluating the efficacy of nisin.

Development of a Modeling Tool To Assess and Reduce Regulatory and Recall Risks for Cold-Smoked Salmon Due to Listeria monocytogenes Contamination.

ABSTRACT: Although public health risk assessments for Listeria monocytogenes (Lm) have been published for various foods, firm-level decision making on interventions targeting Lm involves considerations of both public health and enterprise risks. Smoked seafood is a ready-to-eat product with a high incidence of Lm contamination and has been associated with several recalls. We used cold-smoked salmon as a model product to develop a decision support tool (the regulatory and recall risk [3R] model) to estimate (i) baseline regulatory and recall (RR) risks (i.e., overall risks of a lot sampled and found positive for Lm, e.g., by food regulatory agencies) due to Lm contamination and (ii) the RR risk reduction that can be achieved through interventions with underlying mechanisms such as reducing the prevalence and/or level of Lm and retarding or preventing Lm growth. Given that a set number of samples (e.g., 10) are tested for a given lot, the RR risk equals the likelihood of detecting Lm in at least one sample. Under the baseline scenario, which assumes a 4% Lm prevalence and no interventions, the median predicted RR risk for a given production lot was 0.333 (95% credible interval: 0.288, 0.384) when 10 25-g samples were tested. Nisin treatments, which reduce both the prevalence and initial level of Lm, reduced RR risks in a concentration-dependent manner to 0.109 (0.074, 0.146) with 5 ppm, 0.049 (0.024, 0.083) with 10 ppm, and 0.017 (0.007, 0.033) with 20 ppm. In general, more effective reduction in RR risks can be achieved by reducing Lm prevalence than by retarding Lm growth; the RR risk was reduced to 0.182 (0.153, 0.213) by a 50% prevalence reduction but to only 0.313 (0.268, 0.367) by bacteriostatic growth inhibitors. Sensitivity analysis indicated that prevalence and initial level of Lm and storage temperature have the greatest impact on predicting RR risks, suggesting that reliable data for these parameters will improve model performance.

Growth and survival of aerobic and Gram-negative bacteria on fresh spinach in a Chinese supply chain from harvest through distribution and refrigerated storage.

Spinach is a highly perishable product that degrades over time, including due to bacteria contaminating the product prior to packaging, yet the dynamics of bacterial spoilage and factors that affect it are not well understood. Notably, while China is the top producer of spinach globally, there is limited available microbiological data in the literature for spinach supply chains in China. The overall goal of this foundational study was to establish a baseline understanding of bacterial population dynamics on spinach from harvest to 10 days postprocessing for a Chinese supply chain that includes distribution via traditional grocery (a local physical store) and eCommerce (an online store). To this end, organic spinach samples were collected at different stages in a Chinese supply chain by following the same 3 lots, starting at point-of-harvest through processing and distribution via a local grocery store and eCommerce. After distribution, the same 3 lots were stored at 4 °C with microbiological testing performed on multiple days up to day 10 postprocessing, simulating storage at the point-of-consumer. Results showed aerobic plate counts and total Gram-negative counts did not significantly differ across stages in the supply chain from harvest through processing. However, packaged spinach from the same processing facility and lots, exhibited different patterns in bacterial levels across 0 to 10 days postprocessing, depending on whether it was distributed via the local grocery store or eCommerce. Evaluation of bacterial populations performed on a subset of the packaged spinach samples indicated Gram-negative bacteria, in particular Pseudomonas, were predominant across all days of testing (days 0, 3, and 10 postprocessing), with populations differing at the genus level by day. Overall, this study improves our understanding of the dynamics of bacterial populations on spinach and provides baseline data needed for future studies.

Characterization of Listeria monocytogenes isolated from wildlife in central New York.

BACKGROUND: Listeria monocytogenes (Lm) present in farming soil and food-processing facilities threatens food safety, but little is known about the carriage of Lm by wildlife. OBJECTIVES: We estimated the prevalence of faecal Lm shedding among wildlife admitted to a veterinary medical teaching hospital in central New York and characterized a subset of the Lm isolates. METHODS: Wildlife samples were collected between May 2018 and December 2019. We characterized the Lm isolates by assessing the growth at three temperatures approximating the body temperatures of reptiles (25°C), mammals (37°C), and birds (42°C) and identifying genotypic characteristics related to transmission and virulence. RESULTS: The apparent prevalence of faecal Lm shedding was 5.6% [18/324; 95% confidence interval (CI), 3.3%-8.6%]. Among 13 isolates that represented two lineages and 11 clonal complexes, three and five isolates were grouped into the same SNP clusters with human clinical isolates and environmental isolates, respectively. However, specific SNP difference data showed that Lm from wildlife was generally not closely related (>22 SNP differences) to Lm from human clinical sources and the food-processing environment. While the stress response locus SSI-2 was absent, SSI-1 was found in four isolates. Virulence genes prfA, plcA, hly, mpl, actA, plcB, inlA, inlB, inlC, inlE, inlH, inlJ, and inlK were present, without any premature stop codons, in all isolates. Virulence loci Listeria pathogenicity island 3 (LIPI-3) and LIPI-4, which have been linked to hypervirulence, and inlG were found in four, three, and seven isolates, respectively. CONCLUSIONS: Wildlife represents a potential reservoir for genetically diverse and putatively hypervirulent Lm strains. No statistically significant association between growth parameters and hosts was observed. However, compared to lineage I isolates, lineage II isolates showed significantly (p < 0.05) faster growth at 25°C and significantly slower growth at 42°C, suggesting that wildlife Lm isolates that belong to lineages I and II differ in their ability to grow at 25°C and 42°C.

Development of a Genomics-Based Approach To Identify Putative Hypervirulent Nontyphoidal Salmonella Isolates: Salmonella enterica Serovar Saintpaul as a Model.

While differences in human virulence have been reported across nontyphoidal Salmonella (NTS) serovars and associated subtypes, a rational and scalable approach to identify Salmonella subtypes with differential ability to cause human diseases is not available. Here, we used NTS serovar Saintpaul (S. Saintpaul) as a model to determine if metadata and associated whole-genome sequence (WGS) data in the NCBI Pathogen Detection (PD) database can be used to identify (i) subtypes with differential likelihoods of causing human diseases and (ii) genes and single nucleotide polymorphisms (SNPs) potentially responsible for such differences. S. Saintpaul SNP clusters (n = 211) were assigned different epidemiology types (epi-types) based on statistically significant over- or underrepresentation of human clinical isolates, including human associated (HA; n = 29), non-human associated (NHA; n = 23), and other (n = 159). Comparative genomic analyses identified 384 and 619 genes overrepresented among isolates in 5 HA and 4 NHA SNP clusters most significantly associated with the respective isolation source. These genes included 5 HA-associated virulence genes previously reported to be present on Gifsy-1/Gifsy-2 prophages. Additionally, premature stop codons in 3 and 7 genes were overrepresented among the selected HA and NHA SNP clusters, respectively. Tissue culture experiments with strains representing 4 HA and 3 NHA SNP clusters did not reveal evidence for enhanced invasion or intracellular survival for HA strains. However, the presence of sodCI (encoding a superoxide dismutase), found in 4 HA and 1 NHA SNP clusters, was positively correlated with intracellular survival in macrophage-like cells. Post hoc analyses also suggested a possible difference in intracellular survival among S. Saintpaul lineages. IMPORTANCE Not all Salmonella isolates are equally likely to cause human disease, and Salmonella control strategies may unintentionally focus on serovars and subtypes with high prevalence in source populations but are rarely associated with human clinical illness. We describe a framework leveraging WGS data in the NCBI PD database to identify Salmonella subtypes over- and underrepresented among human clinical cases. While we identified genomic signatures associated with HA/NHA SNP clusters, tissue culture experiments failed to identify consistent phenotypic characteristics indicative of enhanced human virulence of HA strains. Our findings illustrate the challenges of defining hypo- and hypervirulent S. Saintpaul and potential limitations of phenotypic assays when evaluating human virulence, for which in vivo experiments are essential. Identification of sodCI, an HA-associated virulence gene associated with enhanced intracellular survival, however, illustrates the potential of the framework and is consistent with prior work identifying specific genomic features responsible for enhanced or reduced virulence of nontyphoidal Salmonella.

STSRNet: Self-Texture Transfer Super-Resolution and Refocusing Network.

Biomedical microscopy images with high-resolution (HR) and axial information can help analysis and diagnosis. However, obtaining such images usually takes more time and economic costs, which makes it impractical in most scenarios. In this paper, we first propose a novel Self-texture Transfer Super-resolution and Refocusing Network (STSRNet) to reconstruct HR multi-focal plane (MFP) images from a single 2D low-resolution (LR) wide field image without relying on scanning or any special devices. The proposed STSRNet consists of three parts: the backbone module for extracting features, the self-texture transfer module for transferring and fusing features, and the flexible reconstruction module for SR and refocusing. Specifically, the self-texture transfer module is designed for images with self-similarity such as cytological images and it searches for similar textures within the image and transfers to help MFP reconstruction. As for reconstruction module, it is composed of multiple pluggable components, each of which is responsible for a specific focal plane, so as to performs SR and refocusing all focal planes at one time to reduce computation. We conduct extensive experiments on cytological images and the experiments show that MFP images reconstructed by STSRNet have richer details in the axial and horizontal directions than input images. At the same time, the reconstructed MFP images also perform better than single 2D wide field images on high-level tasks. The proposed method provides relatively high-quality MFP images when real MFP images cannot be obtained, which greatly expands the application potential of LR wide-field images. To further promote the development of this field, we released our cytology dataset named RSDC for more researchers to use.

Characterization of Basal Transcriptomes Identifies Potential Metabolic and Virulence-Associated Adaptations Among Diverse Nontyphoidal Salmonella enterica Serovars.

The zoonotic pathogen Salmonella enterica includes >2,600 serovars, which differ in the range of hosts they infect and the severity of disease they cause. To further elucidate the mechanisms behind these differences, we performed transcriptomic comparisons of nontyphoidal Salmonella (NTS) serovars with the model for NTS pathogenesis, S. Typhimurium. Specifically, we used RNA-seq to characterize the understudied NTS serovars S. Javiana and S. Cerro, representing a serovar frequently attributed to human infection via contact with amphibians and reptiles, and a serovar primarily associated with cattle, respectively. Whole-genome sequence (WGS) data were utilized to ensure that strains characterized with RNA-seq were representative of their respective serovars. RNA extracted from representative strains of each serovar grown to late exponential phase in Luria-Bertani (LB) broth showed that transcript abundances of core genes were significantly higher (p<0.001) than those of accessory genes for all three serovars. Inter-serovar comparisons identified that transcript abundances of genes in Salmonella Pathogenicity Island (SPI) 1 were significantly higher in both S. Javiana and S. Typhimurium compared to S. Cerro. Together, our data highlight potential transcriptional mechanisms that may facilitate S. Cerro and S. Javiana survival in and adaptation to their respective hosts and impact their ability to cause disease in others. Furthermore, our analyses demonstrate the utility of omics approaches in advancing our understanding of the diversity of metabolic and virulence mechanisms of different NTS serovars.

Cross-Streams Through the Ventral Posteromedial Thalamic Nucleus to Convey Vibrissal Information.

Whisker detection is crucial to adapt to the environment for some animals, but how the nervous system processes and integrates whisker information is still an open question. It is well-known that two main parallel pathways through Ventral posteromedial thalamic nucleus (VPM) ascend to the barrel cortex, and classical theory suggests that the cross-talk from trigeminal nucleus interpolaris (Sp5i) to principal nucleus (Pr5) between the main parallel pathways contributes to the multi-whisker integration in barrel columns. Moreover, some studies suggest there are other cross-streams between the parallel pathways. To confirm their existence, in this study we used a dual-viral labeling strategy and high-resolution, large-volume light imaging to get the complete morphology of individual VPM neurons and trace their projections. We found some new thalamocortical projections from the ventral lateral part of VPM (VPMvl) to barrel columns. In addition, the retrograde-viral labeling and imaging results showed there were the large trigeminothalamic projections from Sp5i to the dorsomedial section of VPM (VPMdm). Our results reveal new cross-streams between the parallel pathways through VPM, which may involve the execution of multi-whisker integration in barrel columns.

Expansion tomography for large volume tissue imaging with nanoscale resolution.

Expansion microscopy enables conventional diffraction limit microscopy to achieve super-resolution imaging. However, the enlarged tissue lacks an objective lens with sufficient working distance that can image tissues with whole-brain-scale coverage. Here, we present expansion tomography (ExT) to solve this problem. We have established a modified super-absorbent hydrogel (ExT gel) that possesses high mechanical strength and enables serial sectioning. ExT gel enables tissue and cell imaging and is compatible with various fluorescent labeling strategies. Combining with the high-throughput light-sheet tomography (HLTP) system, we have shown the capability of large volume imaging with nanoscale resolution of mouse brain intact neuronal circuits. The ExT method would allow image samples to support super-resolution imaging of intact tissues with virtually unlimited axial extensions.

Chemical Sectioning for Immunofluorescence Imaging.

Immunofluorescence (IF) is a powerful investigative tool in biological research and medical diagnosis, whereas conventional imaging methods are always conflict between speed, contrast/resolution, and specimen volume. Chemical sectioning (CS) is an effective method to overcome the conflict, which works by chemically manipulating the off/on state of fluorescent materials and turning on only the extremely superficial surface fluorescence of tissues to realize the sectioning capacity of wide-field imaging. However, the current mechanism of CS is only applicable to samples labeled with pH-sensitive fluorescent proteins and still cannot fulfill samples immunolabeled with frequently used commercial fluorescent dyes. Here, immunofluorescence chemical sectioning (IF-CS) is described to present an off/on mechanism for Alexa dyes by complexation reactions, allowing CS imaging of IF labeled tissues. IF-CS enables IF freeing from out-of-focus interference in wide-field imaging and satisfying with multicolor imaging. IF-CS demonstrates the utility of the 3D submicron-resolution imaging of large immunolabeled tissues on the wide-field block-face system. IF-CS may remarkably facilitate systematic studies of refined subcellular architectures of endogenous proteins in intact biological systems.

Survival and transcriptomic response of Salmonella enterica on fresh-cut fruits.

Salmonella enterica is frequently implicated in foodborne disease outbreaks associated with fresh-cut fruits. In the U.S., more than one third of fruit-related outbreaks have been linked to two S. enterica serotypes Newport and Typhimurium. Approximately 80% of fruit-related human salmonellosis cases were associated with tomatoes, cantaloupes and cucumbers. In this study, we investigated the population dynamics of S. Newport and S. Typhimurium on fresh-cut tomato, cantaloupe, cucumber and apple under short-term storage conditions. We further compared the transcriptomic profiles of a S. Newport strain on fresh-cut tomato and cantaloupe using high-throughput RNA-seq. We demonstrated that both S. enterica Newport and Typhimurium survived well on various fresh-cut fruit items under refrigeration storage conditions, independent of inoculation levels. However, S. enterica displayed variable survival behaviors on different types of fruits. For example, at 7 d storage, the population of S. enterica reduced less than 0.2 log (p > 0.05) on fresh-cut tomato and cantaloupe, in contrast to ~0.5 log (p < 0.05) on cucumber and apple. RNA-seq analysis suggested that S. enterica mediates its survival on fresh-cut fruits through differentially regulating genes involved in specific carbon utilization and metabolic pathways. Several known bacterial virulence factors (e.g., pag gene) were found to be differentially regulated on fresh-cut tomato and cantaloupe, suggesting a link between the events of food contamination and subsequent human infection. Findings from this study contribute to a better understanding of S. enterica survival mechanisms on fresh-cut produce.

Extending the 3D scanning range of DMD-based scanners for femtosecond lasers.

Digital micromirror devices (DMDs) have shown their potential in 2-photon imaging and microfabrication as diffractive scanners for femtosecond lasers. However, the scanning range of a DMD-based scanner is decreased by the spatial filter (SF) used to block undesired diffraction orders. Instead of an SF, we present a method of introducing and correcting aberration (ICA) to reduce the effects of these undesired diffraction orders. In ICA, aberrations are introduced by optical elements, and only the aberration of the desired diffraction order is corrected by adding a compensatory phase to the scanning phase. The scanning ranges in the y and z directions can be nearly doubled when the SF is removed. We demonstrate that ICA can be conveniently applied to a previously constructed DMD-based 2-photon microscope, and the field of view can be extended at different axial positions.