Development and validation of an improved method for the detection of Salmonella in cinnamon bark and oregano leaves using the adsorbent beta zeolite in the pre-enrichment media.

The detection of Salmonella in spices is challenging due to the presence of antibacterial components. In this study, we evaluated the use of an adsorbent beta zeolite in pre-enrichment media to improve the recovery of Salmonella from cinnamon bark and oregano leaves. Samples (25 g) were spiked with varying levels of S. Montevideo or S. Senftenberg. After 2 weeks of stabilization at RT, betazeolite was added to cinnamon and oregano samples prior to the addition of 225 mL or 475 mL of pre-enrichment media, respectively. Detection sensitivity and rate of the test method were compared to the FDA Bacteriological Analytical Manual (BAM) method which requires the use of 2.5 L pre-enrichment broth. While Salmonella could not be detected in the test method using the reduced volume of pre-enrichment media alone, the addition of beta zeolite resulted in a positivity rate of 62% and 72.6% for cinnamon bark and oregano leaves respectively (all spike levels and both serovars combined). Furthermore, while there were differences in the LOD50 compared to the BAM method, there was no significant difference in the minimum level of detection between the betazeolite and the BAM methods. Our results demonstrate that the use of betazeolite in the pre-enrichment media offers a method with reduced media volumes without compromising on the sensitivity or efficiency of Salmonella detection in cinnamon bark and oregano leaves.

Drying characteristics of mint leaves (Mentha arvensis) dried in a solid desiccant dehumidifier system.

In this present study, solid desiccant-based pressure-swing adsorption (PSA) dehumidifier was developed and the process parameters were optimized to deliver the air continuously at 0.1% relative humidity. Mint (Mentha arvensis) leaves are tested to study the drying characteristics at varied flow rates of dehumidified air in the drying chamber. The initial moisture content of 5.059 g water/g dry matter have been reduced to a safe storage level in 360 min at 0.160 m3/min volume low rate. The effective moisture diffusivity of the mint leaves was found in the range of 2.07534 × 10-11m2/s to 3.45817 × 10-11m2/s. The percentage of retention of ascorbic acid in dried mint leaves is increased by an increase in the volume flow rate of dry air and a maximum of 70.11% is achieved by 0.160 m3/min. The colour measurement and chlorophyll content of the dried samples indicated that the desiccant dehumidified air dryers are suitable for heat sensitive green leafy vegetables.

Mercury in the sediments of freshwater lakes in Ny-Ålesund, Arctic.

Mercury and its speciation in aquatic ecosystems have been assessed globally. Even though previous studies were limited to Arctic freshwater lakes, they are highly significant in the context of the changing climate. The present study is based on sediment samples collected from three Arctic freshwater lakes over a period of 4 years (2015-2018). The samples were analysed for total mercury (THg), methyl mercury (MHg), and various mercury fractions. The observed mean THg and MHg concentrations were 22.23 ng/g and 0.41 ng/g respectively; these values were comparable with those for other Arctic freshwater lakes. The mercury content significantly varied among the years as well as among the lakes. Changes in snowdrift and meltwater inputs, which are the major sources of water for the lakes, may have influenced the sediment mercury content along with geographical location and increased productivity. The results of MHg indicated the susceptibility of lake sediments to methylation. The major fractions observed were the organo-chelated form of mercury, followed by the elemental and water-soluble forms. These results indicate the availability of mercury for methylation. Hence, it is necessary to conduct more studies on the influence of climate change, mercury release through permafrost melting, and atmospheric deposition.

Diet-induced obesity precipitates kidney dysfunction and alters inflammatory mediators in mice treated with Shiga Toxin 2.

Shiga Toxin (Stx)-producing E. coli (STEC) continue to be a prominent cause of foodborne outbreaks of hemorrhagic colitis worldwide, and can result in life-threatening diseases, including hemolytic uremic syndrome (HUS), in susceptible individuals. Obesity-associated immune dysfunction has been shown to be a risk factor for infectious diseases, although few studies have addressed the role of obesity in foodborne diseases. We hypothesized that obesity may affect the development of HUS through an alteration of immune responses and kidney function. We combined diet-induced obese (DIO) and HUS mouse models to look for differences in disease outcome between DIO and wild-type (WT) male and female C57 B l/6 mice. Following multiple intraperitoneal injections with endotoxin-free saline or sublethal doses of purified Stx2, we examined DIO and WT mice for signs of HUS development. DIO mice receiving Stx2 injections lost more body weight, and had significantly higher (p < 0.001) BUN, serum creatinine, and neutrophil counts compared to WT mice or DIO mice receiving saline injections. Lymphocyte counts were significantly (p < 0.05) lower in Stx2-treated obese mice compared to WT mice or saline-treated DIO mice. In addition to increased Stx2-induced kidney dysfunction, DIO mouse kidneys also had significantly increased expression of IL-1α, IL-1ß, IL-6, TNF-α, MCP-1, and KC RNA compared to saline controls (p < 0.05). Serum cytokine levels of IL-6 and KC were also significantly higher in Stx2-treated mice compared to saline controls, but there were no significant differences between the WT and DIO mice. WT and DIO mice treated with Stx2 exhibited significantly higher degrees of kidney tubular dilation and necrosis as well as some signs of tissue repair/regeneration, but did not appear to progress to the full pathology typically associated with human HUS. Although the combined obesity/HUS mouse model did not manifest into HUS symptoms and pathogenesis, these data demonstrate that obesity alters kidney function, inflammatory cells and cytokine production in response to Stx2, and may play a role in HUS severity in a susceptible model of infection.

Negative Differential Resistance Device from Organic/Inorganic Hybrid Nanostructures.

Negative differential resistance device (NDR) fabricated by spin coating of organic/inorganic hybrid nanostructures at room temperature, is reported in the present paper. The coated organic layer is MEH-PPV (poly-[2-methoxy-5-(2'-ethyl-hexyloxy)-1,4-phenylenevinylene] and inorganic layer is ZnO nanoparticles. The device shows negative differential resistance at low voltage and I­V characteristics of the device show multiple peaks at low voltage values. A value of 13 and 4 for the peak- to-valley ratio of current are reported in bi-layer and single layer structures respectively. Depending on the observed NDR signature, operating mechanisms are explored based on carrier (resonant) tunneling process and donor like trap mechanisms. This results show that the MEH-PPV/ZnO thin films gives good performance and is relevant for applications in optoelectronic devices such as a negative differential resistance.

Efficacy and tolerance of papain gel with conventional drilling method: a clinico-microbiological study.

BACKGROUND: Conventional methods of caries removal are commonly associated with pain, fear and discomfort. Chemomechanical methods were introduced to instill a positive dental attitude. Agents like GK-101, Caridex, Carisolv did not prove effective alternatives owing to their high cost, need of special instruments and taste of chlorine. A new chemomechanical agent, Papacarie®, has been introduced to overcome these deficiencies. OBJECTIVE: This study was aimed to compare the effectiveness and tolerance of Papacárie® with the conventional method. METHOD: 25 children with at least two primary teeth with broad cavitated occlusal or cervical lesion were selected. One carious tooth from each patient was randomly treated with each of Papacarie® and conventional drilling method, one after the other. Time taken for caries excavation, child's pain perception, change in anxiety levels, microbial flora and child's preference of treatment were recorded separately for both the methods. RESULT: Although the mean time taken for caries removal by the Papacarie® method was slightly longer (P≯0.05) but it led to reduction in pain and anxiety (p<0.05). The viable bacterial counts were significantly reduced by either of the two methods (P <0�0001). More patients preferred Papacarie® over conventional method of treatment (P<0.05). CONCLUSION: Papacárie® method seems to be a better alternative to conventional method of caries removal.

Detection of Cyclospora cayetanensis in Food and Water Samples: Optimized Protocols for Specific and Sensitive Molecular Methods from a Regulatory Agency Perspective.

Cyclospora cayetanensis is a coccidian parasite of the phylum Apicomplexa that causes cyclosporiasis, a human-specific gastrointestinal disease. Unlike most enteric pathogens, C. cayetanensis does not infect via direct fecal-oral transmission between humans because shed oocysts must be exposed to environmental triggers prior to becoming infectious. The development of specific and sensitive detection methods for C. cayetanensis is crucial to effectively address data gaps and provide regulatory support during outbreak investigations. In this study, new more specific molecular markers for the detection of C. cayetanensis were developed based on updated genomic databases of Apicomplexa mitochondrial sequences. Novel alternative reagents and supplies, as well as optimization protocols, were tested in spiked produce and agricultural water samples. The selected Mit1C primers and probe combined showed at least 13 mismatches to other related species. The new optimized qualitative real-time PCR assay with modifications to sample processing and replacement of discontinued items produced results comparable to the previously validated methods. In conclusion, the new optimized qualitative Mit1C real-time PCR assay demonstrated an increase in its specificity in comparison to other detection methods previously published, while it showed to be robust and as sensitive as the previously validated method at the FDA. This study has also expanded the array of PCR reagents that can be used to detect C. cayetanensis in produce and agricultural water samples and provided several improvements to the method for detection in agricultural water including replacements for discontinued items and a new dialysis filter for water filtration.

Tissue Inhibitor of Metalloproteinase 3 (TIMP3) mutations increase glycolytic activity and dysregulate glutamine metabolism in RPE cells.

OBJECTIVES: Mutations in Tissue Inhibitor of Metalloproteinases 3 (TIMP3) cause Sorsby's Fundus Dystrophy (SFD), a dominantly inherited, rare form of macular degeneration that results in vision loss. TIMP3 is synthesized primarily by retinal pigment epithelial (RPE) cells, which constitute the outer blood-retinal barrier. One major function of RPE is the synthesis and transport of vital nutrients, such as glucose, to the retina. Recently, metabolic dysfunction in RPE cells has emerged as an important contributing factor in retinal degenerations. We set out to determine if RPE metabolic dysfunction was contributing to SFD pathogenesis. METHODS: Quantitative proteomics was conducted on RPE of mice expressing the S179C variant of TIMP3, known to be causative of SFD in humans. Proteins found to be differentially expressed (P < 0.05) were analyzed using statistical overrepresentation analysis to determine enriched pathways, processes, and protein classes using g:profiler and PANTHER Gene Ontology. We examined the effects of mutant TIMP3 on RPE metabolism using human ARPE-19 cells expressing mutant S179C TIMP3 and patient-derived induced pluripotent stem cell-derived RPE (iRPE) carrying the S204C TIMP3 mutation. RPE metabolism was directly probed using isotopic tracing coupled with GC/MS analysis. Steady state [U-13C6] glucose isotopic tracing was preliminarily conducted on S179C ARPE-19 followed by [U-13C6] glucose and [U-13C5] glutamine isotopic tracing in SFD iRPE cells. RESULTS: Quantitative proteomics and enrichment analysis conducted on RPE of mice expressing mutant S179C TIMP3 identified differentially expressed proteins that were enriched for metabolism-related pathways and processes. Notably these results highlighted dysregulated glycolysis and glucose metabolism. Stable isotope tracing experiments with [U-13C6] glucose demonstrated enhanced glucose utilization and glycolytic activity in S179C TIMP3 APRE-19 cells. Similarly, [U-13C6] glucose tracing in SFD iRPE revealed increased glucose contribution to glycolysis and the TCA cycle. Additionally, [U-13C5] glutamine tracing found evidence of altered malic enzyme activity. CONCLUSIONS: This study provides important information on the dysregulation of RPE glucose metabolism in SFD and implicates a potential commonality with other retinal degenerative diseases, emphasizing RPE cellular metabolism as a therapeutic target.

Robust switching characteristics of CdSe/ZnS quantum dot non-volatile memory devices.

In this paper we report Al/CdSe-ZnS core-shell quantum dot/AlOx/CdSe-ZnS core-shell quantum dot/ITO based non-volatile resistive memory devices with an ON/OFF ratio of ~1000. The facile solution processed device exhibited excellent endurance characteristics for 200,000 switching cycles. Retention tests showed good stability for over 20,000 s and the devices are reproducible. A memory operating mechanism is proposed based on charge trapping-detrapping in core-shell quantum dots with AlOx acting as a barrier leading to Coulomb blockade. I-V characteristics of a three terminal device fabricated with the additional terminal wired-out from the middle AlOx layer supports the proposed charge trapping mechanism.

2-Amino-6-methyl-pyridinium 4-hy-droxy-benzoate.

In the title mol-ecular salt, C6H9N2 (+)·C7H5O3 (-), the dihedral angle between the benzene ring and the CO2 group in the anion is 6.1 (2)°. In the crystal, the cation and anion are linked by N-H⋯O and C-H⋯O hydrogen bonds, and the anions are connected by O-H⋯O hydrogen bonds, forming a three-dimensional network.

A comparative study of rectal volume variation in patients with prostate cancer: A tertiary care center study.

INTRODUCTION: Every day variations in rectal filling in prostate cancer radiotherapy can significantly alter the delivered dose distribution from what was intended. The goal of this study was to see if the time of treatment delivery affected the rectal filling. METHODS: This is a retrospective study which included 50 patients with localized prostate cancer treated with volumetric modulated arc therapy (VMAT) to the primary and regional lymph nodes. Cone Beam Computed Tomography (CBCT) image-sets were done for all patient's daily setup verification. The radiation therapist contoured the rectum on all CBCT image sets. The rectal volumes delineated on CBCT and the planning CT image sets were compared. The change in rectal volumes between morning and afternoon treatments were calculated and compared. RESULTS: A total of 1000 CBCT image sets were obtained on 50 patients in the morning and afternoon. The percentage variation of the CBCT rectal volumes over the planning CT scan was 16.57% in the AM group and 24.35% in the PM group. CONCLUSION: The percentage change in rectal volume was significantly lesser in AM group compared to PM group and therefore morning treatments may result in dose distribution that is close to the intended dose distribution. IMPLICATIONS FOR PRACTICE: In prostate cancer radiotherapy our study suggests that a simple technique of changing the time of treatment from afternoon to morning can help to reduce the rectal volume.

Multi-Institutional Clinical Outcomes of Biopsy Gleason Grade Group 5 Prostate Cancers Treated With Contemporary High-Dose Radiation and Long-Term Androgen Deprivation Therapy.

AIMS: This multicentric retrospective study reports long-term clinical outcomes of non-metastatic grade group 5 prostate cancers treated with external beam radiotherapy (EBRT) alone with long-term androgen deprivation therapy (ADT). MATERIALS AND METHODS: Patients treated across 19 institutions were studied. The key endpoints that were evaluated were 5-year biochemical recurrence-free survival (bRFS), metastases-free survival (MFS), overall survival, together with EBRT-related acute and late toxicities. The impact of various prognostic factors on the studied endpoints was analysed using univariate and multivariate analyses. RESULTS: Among the 462 patients, 88% (405) had Gleason 9 disease and 31% (142) had primary Gleason pattern 5. A prostate-specific membrane antigen positron emission tomography-computed tomography scan was used for staging in 33% (153), 80% (371) were staged as T3/T4 and 30% (142) with pelvic nodal disease. The median ADT duration was 24 months; 66% received hypofractionated EBRT and 71.4% (330) received pelvic nodal irradiation. With a median follow-up of 56 months, the 5-year bRFS, MFS and overall survival were 73.1%, 77.4% and 90.5%, respectively. Primary Gleason pattern 5 was associated with worse bRFS, MFS and overall survival with hazard ratios of 0.51 (95% confidence interval 0.35 to 0.73, P < 0.001), 0.64 (95% confidence interval 0.43 to 0.96, P = 0.031) and 0.52 (95% confidence interval 0.28 to 0.97, P = 0.040), respectively, whereas pelvic nodal disease was associated with worse bRFS (hazard ratio 0.67, 95% confidence interval 0.46 to 0.98, P = 0.039) and MFS (hazard ratio 0.56, 95% confidence interval 0.37 to 0.85, P = 0.006). The acute and late radiation-related toxicities were low overall and pelvic nodal irradiation was associated with higher toxicities. CONCLUSION: Contemporary EBRT and long-term ADT led to excellent 5-year clinical outcomes and low rates of toxicity in this cohort of non-metastatic grade group 5 prostate cancers. Primary Gleason pattern 5 and pelvic node disease portends inferior clinical outcomes.

Development and Single Laboratory Evaluation of a Refined and specific Real-time PCR Detection Method, Using Mitochondrial Primers (Mit1C), for the Detection of Cyclospora cayetanensis in Produce.

Regulatory methods for detection of the foodborne protozoan parasite Cyclospora cayetanensis must be specific and sensitive. To that end, we designed and evaluated (in a single laboratory validation) a novel and improved primer/probe combination (Mit1C) for real-time PCR detection of C. cayetanensis in produce. The newly developed primer/probe combination targets a conserved region of the mitochondrial genome of C. cayetanensis that varies in other closely related organisms. The primer/probe combination was evaluated both in silico and using several real-time PCR kits and polymerases against an inclusivity/exclusivity panel comprised of a variety of C. cayetanensis oocysts, as well as DNA from other related Cyclospora spp. and closely related parasites. The new primer/probe combination amplified only C. cayetanensis, thus demonstrating specificity. Sensitivity was evaluated by artificially contaminating cilantro, raspberries, and romaine lettuce with variable numbers (200 and 5) of C. cayetanensis oocysts. As few as 5 oocysts were detected in 75%, 67.7%, and 50% of the spiked produce samples (cilantro, raspberries, and romaine lettuce), respectively, all uninoculated samples and no-template real-time PCR controls were negative. The improved primer/probe combination should prove an effective analytical tool for the specific detection of C. cayetanensis in produce.

Whole genome characterization of thermophilic Campylobacter species isolated from dairy manure in small specialty crop farms of Northeast Ohio.

Introduction: With more public interest in consuming locally grown produce, small specialty crop farms (SSCF) are a viable and growing segment of the food production chain in the United States. Methods: The goal of this study was to investigate the genomic diversity of Campylobacter isolated from dairy manure (n = 69) collected from 10 SSCF in Northeast Ohio between 2018 and 2020. Results: A total of 56 C. jejuni and 13 C. coli isolates were sequenced. Multi-locus sequence typing (MLST) identified 22 sequence types (STs), with ST-922 (18%) and ST-61 (13%) predominant in C. jejuni and ST-829 (62%) and ST-1068 (38%) predominant in C. coli. Interestingly, isolates with similar genomic and gene contents were detected within and between SSCF over time, suggesting that Campylobacter could be transmitted between farms and may persist in a given SSCF over time. Virulence-associated genes (n = 35) involved in the uptake and utilization of potassium and organic compounds (succinate, gluconate, oxoglutarate, and malate) were detected only in the C. jejuni isolates, while 45 genes associated with increased resistance to environmental stresses (capsule production, cell envelope integrity, and iron uptake) were detected only in the C. coli isolates. Campylobacter coli isolates were also sub-divided into two distinct clusters based on the presence of unique prophages (n = 21) or IncQ conjugative plasmid/type-IV secretion system genes (n = 15). Campylobacter coli isolates harbored genes associated with resistance to streptomycin (aadE-Cc; 54%) and quinolone (gyrA-T86I; 77%), while C. jejuni had resistance genes for kanamycin (aph3'-IIIa; 20%). Both species harbored resistance genes associated with ß-lactam (especially, blaOXA-193; up to 100%) and tetracycline (tetO; up to 59%). Discussion/Conclusion: Our study demonstrated that Campylobacter genome plasticity associated with conjugative transfer might provide resistance to certain antimicrobials and viral infections via the acquisition of protein-encoding genes involved in mechanisms such as ribosomal protection and capsule modification.

A longitudinal study to examine the influence of farming practices and environmental factors on pathogen prevalence using structural equation modeling.

The contamination of fresh produce with foodborne pathogens has been an on-going concern with outbreaks linked to these commodities. Evaluation of farm practices, such as use of manure, irrigation water source, and other factors that could influence pathogen prevalence in the farming environment could lead to improved mitigation strategies to reduce the potential for contamination events. Soil, water, manure, and compost were sampled from farms in Ohio and Georgia to identify the prevalence of Salmonella, Listeria monocytogenes (Lm), Campylobacter, and Shiga-toxin-producing Escherichia coli (STEC), as well as Arcobacter, an emerging human pathogen. This study investigated agricultural practices to determine which influenced pathogen prevalence, i.e., the percent positive samples. These efforts identified a low prevalence of Salmonella, STEC, and Campylobacter in soil and water (< 10%), preventing statistical modeling of these pathogens. However, Lm and Arcobacter were found in soil (13 and 7%, respectively), manure (49 and 32%, respectively), and water samples (18 and 39%, respectively) at a comparatively higher prevalence, suggesting different dynamics are involved in their survival in the farm environment. Lm and Arcobacter prevalence data, soil chemical characteristics, as well as farm practices and weather, were analyzed using structural equation modeling to identify which factors play a role, directly or indirectly, on the prevalence of these pathogens. These analyses identified an association between pathogen prevalence and weather, as well as biological soil amendments of animal origin. Increasing air temperature increased Arcobacter and decreased Lm. Lm prevalence was found to be inversely correlated with the use of surface water for irrigation, despite a high Lm prevalence in surface water suggesting other factors may play a role. Furthermore, Lm prevalence increased when the microbiome's Simpson's Diversity Index decreased, which occurred as soil fertility increased, leading to an indirect positive effect for soil fertility on Lm prevalence. These results suggest that pathogen, environment, and farm management practices, in addition to produce commodities, all need to be considered when developing mitigation strategies. The prevalence of Arcobacter and Lm versus the other pathogens suggests that multiple mitigation strategies may need to be employed to control these pathogens.

Intrapulmonary lipopolysaccharide exposure upregulates cytokine expression in the neonatal brainstem.

UNLABELLED: Perinatal inflammation and neonatal sepsis trigger lung and brain injury. We hypothesized that endotoxin exposure in the immature lung upregulates proinflammatory cytokine expression in the brainstem and impairs respiratory control. Lipopolysaccharide (LPS) or saline was administered intratracheally to vagal intact or denervated rat pups. LPS increased brainstem IL-1ß and vagotomy blunted this response. There was an attenuated ventilatory response to hypoxia and increased brainstem IL-1ß expression after LPS. CONCLUSION: Intratracheal endotoxin exposure in rat pups is associated with upregulation of IL-1ß in the brainstem that is vagally mediated and associated with an impaired hypoxic ventilatory response.

2-Amino-5-chloro-pyridinium 4-amino-benzoate.

In the title molecular salt, C(5)H(6)ClN(2) (+)·C(7)H(6)NO(2) (-), the cations and anions are connected by cation-to-anion and anion-to-anion N-H⋯O hydrogen bonds into a three-dimensional network. The dihedral angle between the ring and the CO(2) group in the anion is 7.14 (7)°.

PCDD/Fs, dioxin-like, and non-dioxin like PCBs in the sediments of high Arctic fjords, Svalbard.

Polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) are highly toxic organic compounds, and very few studies on their presence in polar environments have been conducted. This study assessed the concentration and distribution of PCDD/Fs, dioxin-like polychlorinated biphenyls (DL-PCBs), and non-dioxin-like polychlorinated biphenyls in selected fjords of the Svalbard archipelago in Norway. The ∑PCDD/Fs observed for Raudfjorden, Smeerenburgfjorden, Magdalenefjorden, and Kongsfjorden were 22.80 pg/g, 25.65 pg/g, 18.27 pg/g, 33.50 pg/g, and 21.69 pg/g, respectively. The WHO's toxic equivalents values of both ∑PCDD/Fs and ∑DL-PCBs were comparatively higher than those reported in other polar regions. Of the four fjords studied, the sediments from Kongsfjorden exhibited the presence of the most toxic materials, including PCB-126 and PCB-169, of DL-PCBs. More than 80% of the total analysed PCDD/Fs were comprised of highly chlorinated congeners (hexa-to-octa forms). More studies are required to understand the destination and transport of these hazardous pollutants in high Arctic sediments.

Current methodologies and future direction of Campylobacter isolation and detection from food matrices, clinical samples, and the agricultural environment.

Campylobacter spp. are the leading cause of bacterial foodborne infections in both developed and developing countries. The food commodities primarily attributed to campylobacteriosis include raw milk, poultry, seafood, and fresh produce. Furthermore, insects, animal/bird fecal material, and agricultural water have been shown to be the sources of Campylobacter contamination in these commodities. Both established and emerging species of Campylobacter have been recovered from food and environmental sources. Therefore, optimal detection and isolation of Campylobacter spp., including the emerging species, is critical for improved surveillance, prevention, and traceback of Campylobacter outbreaks. This review focuses on the existing variability in Campylobacter enrichment and isolation procedures used by researchers and regulatory agencies worldwide, for various matrices. Additionally, the challenges associated with developing and validating new culture, molecular, and immunological methods for rapid and sensitive Campylobacter detection are discussed.