High Levels Of Toxins Research Articles

Coordination regulation of enhanced performance reveals the tolerance mechanism of Chlamys farreri to azaspiracid toxicity

Azaspiracids (AZAs) are lipid biotoxins produced by the marine dinoflagellates Azadinium and Amphidoma spp. that can accumulate in shellfish and cause food poisoning in humans. However, the mechanisms underlying the tolerance of shellfish to high levels of such toxins remain poorly understood. This study investigated the combined effects of detoxification metabolism and stress-related responses in scallops Chlamys farreri exposed to AZA. Scallops accumulated a maximum of 361.81 μg AZA1 eq/kg and 41.6 % AZA residue remained after 21 days of exposure. A range of AZA2 metabolites, including AZA19, AZA11, and AZA23, and trace levels of AZA2-GST, were detected. Total hemocyte counts significantly increased and ROS levels remained consistently high until gradually decreasing. Immune system activation mediated mitochondrial dysfunction and severe energy deficiency. DEGs increased over time, with key genes CYP2J6 and GPX6 contributing to AZA metabolism. These transcriptome and metabolic results identify the regulation of energy metabolism pathways, including inhibition of the TCA cycle and activation of carbohydrates, amino acids, and lipids. AZA also induced autophagy through the MAPK-AMPK signaling pathways, and primary inhibited PI3K/AKT to decrease mTOR pathway expression. Our results provide additional insights into the resistance of C. farreri to AZA, characterized by re-establishing redox homeostasis toward a more oxidative state.

#2711 Cardiomyocytes exposed to medium conditioned by kidney cells treated with the uremic toxin p-cresyl sulfate undergo changes in cell viability

Abstract Background and Aims Cardiorenal Syndrome (CRS) is a condition involving the heart and the kidneys, where a disturbance in one of these organs can lead to alterations in the other. High levels of uremic toxins (UTs) such as p-Cresyl Sulfate (PCS) can induce renal injury. Furthermore, UTs play a crucial role in inflammatory status and cardiac function. Therefore, it is crucial that studies regarding the influence of uremic toxins in the context of cardiorenal syndrome be conducted. Nevertheless, few studies have been conducted in the context of Acute Kidney Injury (AKI) causing acute damage to the heart, and few mechanisms are still known. The objective of the study was to evaluate the role of PCS in cell viability and inflammatory genes in renal and cardiac cells, to simulate cardiorenal syndrome due to excessive PCS. Method The HEK293 (human epithelial renal cells) and H9c2 (murine cardiomyocyte cells) cell lines were used. HEK293 was treated with concentrations of 0.25 mM, 0.5 mM, 1 mM, and 1.5 mM PCS for 6, 24, and 48 hours. H9c2 was treated with PCS (1.5 mM) and conditioned media from PCS-treated renal cells (1.5 mM) for 6 and 24 hours. MTT assays, growth curve analysis, and Hoechst-PI staining were performed. Gene expression was evaluated by RT-qPCR. ANOVA followed by Bonferroni post-test was conducted, considering P < 0.05 as significant. Results PCS treatment led to a time and dose-dependent decrease in cell viability. The cell growth curve indicated a negative impact across time on renal cell proliferation at all different PCS concentrations. Hoechst-PI assay revealed increased cell death in the treatment with PCS (1.5 mM) for 24 hours. There was no change in the gene expression of IL-6, IL-1β, Bax, and Bcl-2 in renal cells. Cardiac cell viability decreased both after direct PCS treatment (85.3%) and exposure to PCS-conditioned media from renal cells for 24 hours (87.3%). Conclusion PCS reduces renal cell viability and inhibits cell proliferation without affecting the inflammatory status and apoptosis signaling within the evaluated time frames. Cardiac cell viability decreased after PCS treatment, either directly or indirectly, suggesting a cardiotoxic effect of PCS, independent of changes in renal cells.

Synchronic distribution of the dinoflagellate Protoceratium reticulatum and yessotoxins in a high stratified fjord system: Tidal or light modulation?

Protoceratium reticulatum is the main yessotoxin-producer along the Chilean coast. Thus far, the yessotoxin levels recorded in this region have not posed a serious threat to human health. However, a bloom of P. reticulatum during the austral summer of 2022 caused the first ban of shellfish collection, due to the high toxin levels. A bloom of P. reticulatum during the austral summer of 2020 allowed an evaluation of the fine-scale distribution of the dinoflagellate during a tidal cycle. High-resolution measurements of biophysical properties were carried out in mid-summer (February 18–19) at a fixed sampling station in Puyuhuapi Fjord, Chilean Patagonia, as part of an intensive 24-h biophysical experiment to monitor the circadian distributions of P. reticulatum vegetative cells and yessotoxins. High P. reticulatum cell densities (>20 × 103 cells L–1) were found in association with a warmer (14.5–15 °C) and estuarine (23.5–24.5 g kg–1) sub-surface water layer (6–8 m). P. reticulatum cell numbers and yessotoxins followed a synchronic distribution pattern consistent with the excursions of the pycnocline. Nevertheless, the surface aggregation of the cells was modulated by the light cycle, suggesting daily vertical migration. The yessotoxin content per P. reticulatum cell ranged from 9.4 to 52.2 pg. This study demonstrates both the value of fine-scale resolution measurements of biophysical properties in a highly stratified system and the potential ecosystem impact of P. reticulatum strains producing high levels of yessotoxins.

The Impact of Farm and Industrial Feed Waste on the Safety Parameters of Tenebrio molitor Larvae

The rising global demand for animal-based food has an increasingly detrimental ecological impact, exacerbated by significant food waste (approximately one-third of all food). This research aimed to analyze the possibility of changing the usually balanced feed with sustainable alternatives that remain as a by-product of the production of farms, grain processing, and breweries, thus promoting the sustainability of agriculture. The mealworm larvae were reared on different substrates: (1) agar-agar gels, wheat bran, and brewer’s yeast, (2) carrots, wheat bran, and brewer’s yeast, (3) sprouted potatoes, wheat bran, and brewer’s yeast, and (4) carrots, brewers’ spent grain and brewer’s yeast. For analysis, the frozen larvae were lyophilized and tested for chemical safety in three accredited laboratories. The results have shown that all tested samples had lower levels of pesticides than the detection limit. In scientific literature, we didn’t find studies on polycyclic aromatic hydrocarbons (PAH). In our study, we found PAH in the substrate and these toxins, as our study shows, can also enter the larvae, but no significant accumulation was observed (sum of benzo(a)pyrene, benz(a)anthracene, benzo(b)fluoranthene and chrysene was 0.0007 mg/kg). Furthermore, the total content of PAH, benzo(a)pyrene and histamine did not exceed recommended levels. We have noticed that the highest concentration of heavy metals (e.g., chromium—1.45 ± 0.02) was found in the sample with the brewer’s by-products. While numerous studies utilize plant-derived by-products, the accumulation of glycoalkaloids has not been explored. Among the all glycoalkaloids (tomatidin, tomatine, α-solanine, α-chaconine and solanidin), amounts of α-solanine and α-chaconine were the highest, detected in the sample with sprouted potatoes (175.12 ± 0.21 and 139.32 ± 0.32 mg/kg, respectively). The amount of total putrescine, tyramine, spermine, and spermidine in mealworm larvae was statistically higher compared to the amount detected in the substrate, and histamine level-on the contrary, was statistically significantly lower compared to the amount detected in the substrate. Considering the amount of toxic substances found in the substrate from the by-products, we can assume that mealworms did not accumulate high levels of toxins, which would violate regulations.

B-216 Development of a Point-of-care Multiplexed Room Temperature Stable Environmental Toxin Test

Abstract Background Hememics™ Biosensor System (HBS) is a platform technology enabling antigen and molecular testing in the field. The system consists of a reader, HemBox™, and a biosensing chip, HemChip™. The HemBox™ is a battery-operated, cell phone size with Bluetooth and WIFI module that can be integrated into the Health Management System. The biosensing chips can be programed with bio-receptors to detect pathogens in less than 5 min. This study presents performance characteristics of HBS as a rapid POC to detect toxins from environmental samples. Methods The testing protocol consists of insert a HemChip™ into the HemBox™ and mixing the sample with the buffer and apply a drop of this mixture to the chip. In less than 5 min, results are displayed on the screen of HemBox™ or transmitted to user’s cellphone via Bluetooth. Samples were prepared by spiking solutions of mud, serum, vegetable wash, and milk with Staphylococcal enterotoxin B (SEB) and ricin. Results HemChip™ was functionalized to recognize Ricin and SEB targets. For testing, these targets were mixed in various matrix. To achieve statistical significance, each sample tested represented a minimum of 10 data points. Conclusion Hememics™ Biosensor System (HBS) is designed to be the ultimate Portable Lab Diagnostics for field use. The ability to test for environmental toxins in the field is significant because it allows for quick and on-site detection of potentially harmful substances. Our biosensor system also eliminates the need for sample processing, collection and transportation, which can take significant time and resources. The results of field testing can be available almost immediately, allowing for prompt action to be taken in the case of high toxin levels.

Bioengineered Kidney Tubules Efficiently Clear Uremic Toxins in Experimental Dialysis Conditions.

Patients with end-stage kidney disease (ESKD) suffer from high levels of protein-bound uremic toxins (PBUTs) that contribute to various comorbidities. Conventional dialysis methods are ineffective in removing these PBUTs. A potential solution could be offered by a bioartificial kidney (BAK) composed of porous membranes covered by proximal tubule epithelial cells (PTECs) that actively secrete PBUTs. However, BAK development is currently being hampered by a lack of knowledge regarding the cytocompatibility of the dialysis fluid (DF) that comes in contact with the PTECs. Here, we conducted a comprehensive functional assessment of the DF on human conditionally immortalized PTECs (ciPTECs) cultured as monolayers in well plates, on Transwell® inserts, or on hollow fiber membranes (HFMs) that form functional units of a BAK. We evaluated cell viability markers, monolayer integrity, and PBUT clearance. Our results show that exposure to DF did not affect ciPTECs' viability, membrane integrity, or function. Seven anionic PBUTs were efficiently cleared from the perfusion fluid containing a PBUTs cocktail or uremic plasma, an effect which was enhanced in the presence of albumin. Overall, our findings support that the DF is cytocompatible and does not compromise ciPTECs function, paving the way for further advancements in BAK development and its potential clinical application.

Impact of Drought and Wildfires in Recent Trends of Diarrhetic Shellfish Toxins in Cockles from Northwest Portugal and Its Similarities with Sardine Stock Trends in the Period 2001–2022

In Portugal, high levels of diarrhetic shellfish toxins (DSTs) originating from Dinophysis spp. are accumulated in bivalves, mainly on the northwest coast (NWC). The occurrence of DSTs in spring and early summer is positively related to precipitation. A decrease in average precipitation after 2003 led to a reduction in DSTs after 2008. However, the accumulation of DSTs in the NWC increased in the period 2020–2022. The hydrological year of 2022 was marked by extreme drought. In cockles from the endemic area of Ria de Aveiro, 37.3% of the weeks tested presented samples above the regulatory limit (RL). The previous record was 31.9% in 2005, also an extremely dry year. The average percentage of weeks above the RL in severe drought years surpassed both the low and high precipitation years. In severe dry years, toxicity was dominated mainly by D. acuta toxins. While D. acuminata grows after the abundant river discharges in spring, D. acuta is capable of growing in summer during upwelling favourable conditions. In the last two decades, extensive forest wildfires were recurrent, but the area burnt reduced sharply after 2017 following tighter fire control measures. Low levels of DSTs or low percentage of weeks with cockles above the RL were related to high burnt areas in the previous year. The recent increase in 2020–2022 of DSTs, a planktonic biomarker in bivalves, was also coincident with the similar temporal increase in either the recruitment or the biomass of Sardina pilchardus, a planktivorous fish in decline after 2005/2006. Both low sardine recruitment and biomass were coincident with low precipitation or high burnt areas. Wildfires degrade coastal water quality, as seems reflected both in the cockle’s DST and the sardine stock time series.

Analysis of dynamic evolution process of the N-player division of labor game model

This paper investigates a three-strategy (cooperators, toxin producers, and cheaters) N-player division of labor game in bacterial populations. We construct the replicator equation to discuss the evolution of the frequency of the three strategies. Firstly, we prove that the interior equilibrium is always unstable, the three strategies cannot coexist. Secondly, according to Sotomayor’s theorem, the system undergoes transcritical bifurcation. Furthermore, the sensitivity of the two-dimensional evolutionary state diagrams to the third parameter (toxin rate, absorption rate, toxin quantity, etc) is analyzed. In summary, high toxicity rates, high levels of toxins, and low levels of competition tend to promote cooperation. All players choose to perform the task, and the cheater disappears. When the absorption rate of cooperators is high enough, only cooperators exist in the population over time. When the absorption rate of the cooperator is low, and the absorption rate of the toxin producer is greater than the threshold, the cooperator and the toxin producer coexist. All players perform the task. Finally, the triangle diagrams and three-dimensional diagrams are presented, which show the initial conditions of the three strategies also affect the dynamic results. As the amount of toxin increases, the range of players who choose to perform tasks widens.

Impact of nitrogen and phosphorus fertilizers on Cry1Ac protein contents in transgenic cotton.

Application of different fertilizers to check the efficiency of expression of Bt (Bacillus thuringiensis) gene in one of the leading commercialized crops (cotton) against Lepidopteran species is of great concern. The expression of Cry protein level can be controlled by the improvement of nutrients levels. Therefore, the myth of response of Cry toxin to different combinations of NP fertilizers was explored in three Bt cotton cultivars. Combinations include three levels of nitrogen and three levels of phosphorus fertilizers. Immunostrips and Cry gene(s) specific primer based PCR (Polymerase Chain Reaction) analysis were used for the presence of Bt gene that unveiled the presence of Cry1Ac gene only. Further, the ELISA (enzyme-linked immunosorbent assay) kit was used to quantify the expression of Cry1Ac protein. Under various NP fertilizers rates, the level of toxin protein exhibited highly significant differences. The highest toxin level mean was found to be 2.3740 and 2.1732 µg/g under the treatment of N150P75 kg ha-1 combination while the lowest toxin level mean was found to be 0.9158 and 0.7641 µg/g at the N50P25 kg ha-1 level at 80 and 120 DAS (Days After Sowing), respectively. It was concluded from the research that the usage of NP fertilizers has a positive relation with the expression of Cry1Ac toxin in Bt cotton. We recommend using the N150P50 kg ha-1 level as the most economical and practicable fertilizer instead of the standard dose N100P50 kg ha-1 to get the desired level of Cry1Ac level for long lasting plant resistance (<1.5). The revised dose of fertilizer may help farmers to avoid the cross-resistance development in contradiction of insect pests.

Indoxyl sulfate impairs erythropoiesis at BFU-E stage in chronic kidney disease

Chronic kidney disease (CKD) is a global health condition characterized by a progressive deterioration of kidney function. It is associated with high serum levels of uremic toxins (UT), such as Indoxyl Sulfate (IS), which may participate in the genesis of several uremic complications. Anemia is one of the major complications in CKD patients that contribute to cardiovascular disease, increase morbi-mortality, and is associated with a deterioration of kidney failure in these patients. Our study aimed to characterize the impact of IS on CKD-related erythropoiesis. Using cellular and pre-clinical models, we studied cellular and molecular effects of IS on the growth and differentiation of erythroid cells. First, we examined the effect of clinically relevant concentrations of IS (up to 250 μM) in the UT7/EPO cell line. IS at 250 μM increased apoptosis of UT7/EPO cells at 48 h compared to the control condition. We confirmed this apoptotic effect of IS in erythropoiesis in human primary CD34+ cells during the later stages of erythropoiesis. Then, in IS-treated human primary CD34+ cells and in a (5/6 Nx) mice model, a blockage at the burst-forming unit-erythroid (BFU-E) stage of erythropoiesis was also observed. Finally, IS deregulates a number of erythropoietic related genes such as GATA-1, Erythropoietin-Receptor (EPO-R), and β-globin. Our findings suggest that IS could affect cell viability and differentiation of erythroid progenitors by altering erythropoiesis and contributing to the development of anemia in CKD.

Palytoxin-Analogues Accumulation in Natural Mussel Banks during an Ostreopsis cf. ovata Bloom

Intense blooms of the toxic dinoflagellate Ostreopsis producing palytoxin (PlTX) analogs, mainly ovatoxins (OVTXs), have been a recurrent phenomenon along several Mediterranean coasts. Although there is evidence of seafood contamination by these toxins, the dynamics of their bioaccumulation during Ostreopsis blooms is not yet clear. Toxin concentrations in wild mussels at two sites in the Conero Riviera, along the northern Adriatic Sea (Portonovo and Passetto), were analyzed from August to October 2021, throughout an Ostreopsis cf. ovata bloom, to investigate their relationships with the bloom pattern and abundances. Contaminated mussels showed the typical toxin profile dominated by OVTX-a and -b, with levels lower than the threshold considered unsafe for human consumption (30 µg PlTX-equivalent kg−1 soft tissue). The toxin content reached 36.4 µg PlTX kg−1 soft tissue only in a single sampling date. A clear correlation between the bioaccumulation of OVTXs in mussels and the abundance of Ostreopsis was observed. Our results highlighted, however, that bioaccumulation in the mussels is not affected merely by the abundance of toxic microalgae, since higher toxin levels were recorded at Portonovo, where the cell abundances were lower than at Passetto. The results from this study indicate that the Italian guidelines for the management of Ostreopsis blooms in bathing waters are also useful in managing the risks of human intoxication through ingestion, as mussel contamination was detected only during the alert phase (10,000–30,000 cells L−1).

Dynamics of Deoxynivalenol and Nivalenol Glucosylation in Wheat Cultivars Infected with Fusarium culmorum in Field Conditions─A 3 Year Study (2018-2020).

Fusarium head blight (FHB) caused by pathogenic species of Fusarium fungi is one of the most important diseases of cereal plants and a factor contributing to losses in plant production. The growth of FHB-associated species is often accompanied by biosynthesis of secondary metabolites─mycotoxins, which serve as a virulence factor. The aim of the study was to evaluate the ratios between deoxynivalenol (DON) and nivalenol (NIV) and their derivatives in the ears of six cultivars of winter wheat with varying resistance to FHB, taking into account a range of factors (weather conditions, location, cultivar, and year) after inoculation with Fusarium culmorum, during a 3 year field experiment, 2018-2020. The presence of toxins in the ears was measured within 21 days of inoculation. The toxins were found in the ears as soon as on the third day from the start of the experiment, whereas relative humidity higher than 80% was a decisive factor for FHB incidence. All wheat cultivars showed the ability to biotransform DON and NIV present in the ears to glucosides, that is, deoxynivalenol-3-glucoside (DON-3G) and nivalenol-3-glucoside (NIV-3G). The levels of these metabolites showed significant correlation with the levels of their basic analogues. In most cases, higher levels of DON and NIV in wheat ears and higher levels of their metabolites were observed, but the relative levels of DON-3G/DON and NIV-3G/NIV at relatively high levels of toxins were lower compared to the ear samples with relatively low toxin levels. The presented results are the first studies, which systematically correlate a variety of wheat cultivars with their extent to glucosylate trichothecenes.

The Occurrence, Distribution, and Toxicity of High-Risk Ciguatera Fish Species (Grouper and Snapper) in Kiritimati Island and Marakei Island of the Republic of Kiribati.

Ciguatera is one of the most widespread food poisonings caused by the ingestion of fish contaminated by ciguatoxins (CTXs). Snapper and grouper with high palatable and economic value are the primary food source and fish species for exportation in the Republic of Kiribati, but they are highly suspected CTX-contaminated species due to their top predatory characteristics. In this study, 60 fish specimens from 17 species of snappers and groupers collected from the Kiritimati Island and Marakei Island of the Republic of Kiribati were analyzed using mouse neuroblastoma (N2a) assay and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to determine Pacific CTX-1, -2 and -3 (P-CTX-1, -2 and -3). The LC-MS/MS results show that CTXs were detected in 74.5% of specimens from Marakei Island and 61.5% of specimens from Kiritimati Island. The most toxic fish Epinephelus coeruleopunctatus from Marakei Island and Cephalopholis miniata from Kiritimati Island were detected as 53-fold and 28-fold P-CTX-1 equivalents higher than the safety level of 10 pg/g P-CTX-1 equivalents, respectively. CTX levels and composition profiles varied with species and location. The N2a results suggested that fish specimens also contain high levels of other CTX-like toxins or sodium channel activators. The distribution patterns for ciguatoxic fish of the two islands were similar, with fish sampled from the northwest being more toxic than the southwest. This study shows that groupers and snappers are high-risk species for ciguatera in the Republic of Kiribati, and these species can further be used as indicator species in ciguatera endemic areas for risk assessment.

Comparison of laxative effects of fermented soybeans (Cheonggukjang) containing toxins and biogenic amines against loperamide-induced constipation mouse model.

BACKGROUND/OBJECTIVES Cheonggukjang is a traditional fermented soybean paste with significant health-promoting effects. On the other hand, there have been insufficient studies on the safety and efficacy of Cheonggukjang, which is produced using traditional methods containing toxins and biogenic amines (BAs). This study compared the laxative effect of Cheonggukjang, containing high or low levels of toxins and BAs (HTBC or LTBC) in a loperamide (Lop)-induced constipation mouse model.MATERIALS/METHODSTo induce constipation, Lop (5 mg/kg) was administered orally to ICR mice twice a day for 4 days, and the dose was increased to 8 mg/kg after a 3-day rest period. Cheonggukjang (500 mg/kg, HTBC, or LTBC respectively) was administered for four weeks before the Lop treatment.RESULTSThe number of stools, fecal weight, water contents, gastrointestinal transit, and histological alterations were recovered significantly in the HTBC or LTBC groups. HTBC and LTBC administration did not induce significant changes in body weight, dietary intake, and behavior. The opioid-receptor downstream signaling pathway in colon tissues was also evaluated. The c-Kit, stem cell kinase, and mitogen-activated protein kinases subfamilies, including extracellular signal-regulated kinase 1/2, c-Jun N-terminal kinases, and p38, were all downregulated in the HTBC or LTBC-administered mice colon compared to the Lop group.CONCLUSIONThese results show that Cheonggukjang, containing high levels of toxins and BAs, have a similar laxative effect in a mouse model of Lop-induced constipation.

Host Plant Species Mediates Impact of Neonicotinoid Exposure to Monarch Butterflies.

Simple SummaryNeonicotinoids are the most widely used insecticides in North America and many studies document the negative effects of neonicotinoids on bees. Monarch butterflies are famous for their long-distance migrations, and for their ability to sequester toxins from their milkweed host plants. The neonicotinoids imidacloprid and clothianidin were suggested to correlate with declines in North American monarchs. We examined how monarch development, survival, and flight were affected by exposure to neonicotinoids, and how these effects depend on milkweed host plant species that differ in their cardenolide toxins. Monarch survival and flight were unaffected by low and intermediate neonicotinoid doses. At the highest dose, neonicotinoids negatively affected monarch pupation and survival, for caterpillars that fed on the least toxic milkweed. Monarchs fed milkweed of intermediate toxicity experienced moderate negative effects of high insecticide doses. Monarchs fed the most toxic milkweed species had no negative consequences associated with neonicotinoid treatment. Our work shows that monarchs tolerate low neonicotinoid doses, but experience detrimental effects at higher doses, depending on milkweed species. To our knowledge, this is the first study to show that host plant species potentially reduce the residue of neonicotinoid insecticides on the leaf surface, and this phenomenon warrants further investigation.Neonicotinoids are the most widely used insecticides in North America. Numerous studies document the negative effects of neonicotinoids on bees, and it remains crucial to demonstrate if neonicotinoids affect other non-target insects, such as butterflies. Here we examine how two neonicotinoids (imidacloprid and clothianidin) affect the development, survival, and flight of monarch butterflies, and how these chemicals interact with the monarch’s milkweed host plant. We first fed caterpillars field-relevant low doses (0.075 and 0.225 ng/g) of neonicotinoids applied to milkweed leaves (Asclepias incarnata), and found no significant reductions in larval development rate, pre-adult survival, or adult flight performance. We next fed larvae higher neonicotinoid doses (4–70 ng/g) and reared them on milkweed species known to produce low, moderate, or high levels of secondary toxins (cardenolides). Monarchs exposed to the highest dose of clothianidin (51–70 ng/g) experienced pupal deformity, low survival to eclosion, smaller body size, and weaker adult grip strength. This effect was most evident for monarchs reared on the lowest cardenolide milkweed (A. incarnata), whereas monarchs reared on the high-cardenolide A. curassavica showed no significant reductions in any variable measured. Our results indicate that monarchs are tolerant to low doses of neonicotinoid, and that negative impacts of neonicotinoids depend on host plant type. Plant toxins may confer protective effects or leaf physical properties may affect chemical retention. Although neonicotinoid residues are ubiquitous on milkweeds in agricultural and ornamental settings, commonly encountered doses below 50 ng/g are unlikely to cause substantial declines in monarch survival or migratory performance.

Urinary levels of dimethoate, bisphenol A and benzo[a]pyrene in first-year students of Hohai University from different geographical regions

BackgroundThe objective of this study was to detect the urinary levels of dimethoate, benzo(a) pyrene (BaP), and bisphenol A (BPA) in first-year Hohai University students with different geographic origins.MethodsFirst-morning urine samples were collected from 540 healthy freshmen aged 17 to 19 years. Chemical levels were measured using β-glucuronidase hydrolysis followed by a high-performance liquid chromatography-tandem mass spectrometry-based method. Geometric means (GMs) of these three chemicals are presented by body mass index (BMI) and location in a volume-based and creatinine-standardized way.ResultsGM concentrations of omethoate, BPA and 3-OHBaP were 9.47 μg/L (10.80 μg/g creatinine), 3.54 μg/L (4.04 μg/g creatinine) and 0.34 ng/L (0.39 ng/g creatinine), respectively. The GM concentration of omethoate in males was significantly higher than that in females. The individuals with a BMI higher than 23.9 had higher GM concentrations of omethoate, BPA, and 3-OHBaP. The inhabitants of Southwest China had significantly lower GM concentrations of omethoate, BPA, and 3-OHBaP than those who lived in other locations in China.ConclusionThe average level of environmental chemical accumulation in freshmen is lower in Southwest China and differs in youth who live in different regions. In addition, obesity is correlated with higher toxin levels in youth.

Development and validation of a targeted LC-MS/MS quantitation method to monitor cell culture expression of tetanus neurotoxin during vaccine production

The tetanus neurotoxin (TeNT) is one of the most toxic proteins known to man, which prior to the use of the vaccine against the TeNT producing bacteria Clostridium tetani, resulted in a 20% mortality rate upon infection. The clinical detrimental effects of tetanus have decreased immensely since the introduction of global vaccination programs, which depend on sustainable vaccine production. One of the major critical points in the manufacturing of these vaccines is the stable and reproducible production of high levels of toxin by the bacterial seed strains. In order to minimize time loss, the amount of TeNT is often monitored during and at the end of the bacterial culturing. The different methods that are currently available to assess the amount of TeNT in the bacterial medium suffer from variability, lack of sensitivity, and/or require specific antibodies. In accordance with the consistency approach and the three Rs (3Rs), both aiming to reduce the use of animals for testing, in-process monitoring of TeNT production could benefit from animal and antibody-free analytical tools.In this paper, we describe the development and validation of a new and reliable antibody free targeted LC-MS/MS method that is able to identify and quantify the amount of TeNT present in the bacterial medium during the different production time points up to the harvesting of the TeNT just prior to further upstream purification and detoxification. The quantitation method, validated according to ICH guidelines and by the application of the total error approach, was utilized to assess the amount of TeNT present in the cell culture medium of two TeNT production batches during different steps in the vaccine production process prior to the generation of the toxoid. The amount of TeNT generated under different physical stress conditions applied during bacterial culture was also monitored.

High Serum Levels of Toxin A Correlate with Disease Severity in Patients with Clostridioides difficile Infection.

Cloistridioides difficile (CD) represents a major public healthcare-associated infection causing significant morbidity and mortality. The pathogenic effects of CD are mainly caused by the release of two exotoxins into the intestine: toxin A (TcdA) and toxin B (TcdB). CD infection (CDI) can also cause toxemia, explaining the systemic complications of life-threatening cases. Currently, there is a lack of sensitive assays to detect exotoxins circulating in the blood. Here, we report a new semi-quantitative diagnostic method to measure CD toxins serum levels. The dot-blot assay was modified to separately detect TcdA and TcdB in human serum with a limit of detection at the pg/mL levels. TcdA and TcdB concentrations in the plasma of 35 CDI patients were measured at the time of CDI diagnosis and at the fourth and tenth day after CDI diagnosis and initiation of anti-CDI treatment. TcdA and TcdB levels were compared to those determined in nine healthy blood donors. Toxemia was detected in the plasma of 33 out of the 35 CDI cases. We also assessed the relationship between TcdA serum levels and CDI severity, reporting that at the time of CDI diagnosis the proportion of severe CDI cases with a TcdA serum level > 60 pg/µL was higher than in mild CDI cases (29.4% versus 66.6%, p = 0.04). In conclusion, data reported here demonstrate for the first time that toxemia is much more frequent than expected in CDI patients, and specifically that high serum levels of TcdA correlate with disease severity in patients with CDI.

Protecting children from tobacco-related harm in private vehicles

Protecting children from tobacco-related harm in private vehicles

Polyketide synthase genes and molecular trade-offs in the ichthyotoxic species Prymnesium parvum

Prymnesium parvum is a bloom forming haptophyte that has been responsible for numerous fish kill events across the world. The toxicity of P. parvum has been attributed to the production of large polyketide compounds, collectively called prymnesins, which based on their structure can be divided into A-, B- and C-type. The polyketide chemical nature of prymnesins indicates the potential involvement of polyketide synthases (PKSs) in their biosynthesis. However, little is known about the presence of PKSs in P. parvum as well as the potential molecular trade-offs of toxin biosynthesis. In the current study, we generated and analyzed the transcriptomes of nine P. parvum strains that produce different toxin types and have various cellular toxin contents. Numerous type I PKSs, ranging from 37 to 109, were found among the strains. Larger modular type I PKSs were mainly retrieved from strains with high cellular toxin levels and eight consensus transcripts were present in all nine strains. Gene expression variance analysis revealed potential molecular trade-offs associated with cellular toxin quantity, showing that basic metabolic processes seem to correlate negatively with cellular toxin content. These findings point towards the presence of metabolic costs for maintaining high cellular toxin quantity. The detailed analysis of PKSs in P. parvum is the first step towards better understanding the molecular basis of the biosynthesis of prymnesins and contributes to the development of molecular tools for efficient monitoring of future blooms.