RESUMO
The use of lipid-formulated RNA vaccines for cancer or COVID-19 is associated with dose-limiting systemic inflammatory responses in humans that were not predicted from preclinical studies. Here, we show that the 'interleukin 1 (IL-1)-interleukin 1 receptor antagonist (IL-1ra)' axis regulates vaccine-mediated systemic inflammation in a host-specific manner. In human immune cells, RNA vaccines induce production of IL-1 cytokines, predominantly IL-1ß, which is dependent on both the RNA and lipid formulation. IL-1 in turn triggers the induction of the broad spectrum of pro-inflammatory cytokines (including IL-6). Unlike humans, murine leukocytes respond to RNA vaccines by upregulating anti-inflammatory IL-1ra relative to IL-1 (predominantly IL-1α), protecting mice from cytokine-mediated toxicities at >1,000-fold higher vaccine doses. Thus, the IL-1 pathway plays a key role in triggering RNA vaccine-associated innate signaling, an effect that was unexpectedly amplified by certain lipids used in vaccine formulations incorporating N1-methyl-pseudouridine-modified RNA to reduce activation of Toll-like receptor signaling.
Assuntos
Inflamação , Proteína Antagonista do Receptor de Interleucina 1 , Interleucina-1 , Animais , COVID-19 , Inflamação/imunologia , Inflamação/metabolismo , Proteína Antagonista do Receptor de Interleucina 1/genética , Proteína Antagonista do Receptor de Interleucina 1/imunologia , Interleucina-1/genética , Interleucina-1/imunologia , Lipídeos , Camundongos , RNA , Vacinas Sintéticas , Vacinas de mRNA/efeitos adversos , Vacinas de mRNA/metabolismoRESUMO
An effective analytical method was optimized for residues including chlorpyrifos-methyl, deltamethrin, fenoxanil, thiobencarb and fludioxonil in mealworms, the larval form of Tenebrio molitor. They are listed for pest control during wheat cultivation and can be found in wheat-bran feed for growing mealworms in South Korea. Analytes were extracted using acetonitrile and salt packet. Four clean-up methods ((1) MgSO4 + 25 mg PSA + 25 mg C18; (2) MgSO4 + 50 mg PSA + 50 mg C18; (3) EMR-lipidTM tube; and (4) 10 mL n-hexane) were investigated and the method (1) was selected due to its robustness. Low-temperature precipitation of fat and proteins improved the recoveries. Recoveries from the Method (1) were satisfying with 70-120% with <20% relative SD at a spiking level of 0.01 mg/kg. With the simultaneous sample preparation, fenoxanil, thiobencarb and fludioxonil were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) and chlorpyrifos-methyl and deltamethrin by gas chromatography tandem mass spectrometry (GC-MS/MS). Quantification limits for LC-MS/MS and GC-MS/MS were 0.5 and 2.5 µg/L, respectively. No pesticides of interest were detected in 30 real samples collected across the nation. However, the data can be provided for establishing maximum residue limits for the pesticides in mealworms in response to the positive list system.
Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Resíduos de Praguicidas/análise , Espectrometria de Massas em Tandem/métodos , Tenebrio/química , Animais , Clorpirifos/análogos & derivados , Clorpirifos/análise , Clorpirifos/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Imidazóis/análise , Imidazóis/isolamento & purificação , Larva/química , Larva/metabolismo , Limite de Detecção , Extração Líquido-Líquido , Nitrilas/análise , Nitrilas/isolamento & purificação , Resíduos de Praguicidas/isolamento & purificação , Piretrinas/análise , Piretrinas/isolamento & purificação , Tenebrio/crescimento & desenvolvimento , Tenebrio/metabolismoRESUMO
Tenebrio molitor larvae (mealworm) is an edible insect and is considered a future food. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), a novel method for simultaneous analysis of 353 target analytes was developed and validated. Various sample preparation steps including "quick, easy, cheap, effective, rugged, and safe" (QuEChERS) extraction conditions, number of acetonitrile-hexane partitions, and dispersive-solid phase extraction (dSPE) sorbents were compared, and the optimal conditions were determined. In the established method, 5 g of homogenized mealworms was extracted with acetonitrile and treated with QuEChERS EN 15662 salts. The crude extract was subjected to three rounds of acetonitrile-hexane partitioning, and the acetonitrile layer was cleaned with C18 dSPE. The final solution was matrix-matched and injected into LC-MS/MS (2 µL). For target analytes, the limits of quantitation (LOQs) were ≤10 µg/kg, and the correlation coefficient (r2) of calibration was >0.990. In recovery tests, more than 90% of the pesticides showed an excellent recovery range (70-120%) with relative standard deviation (RSD) ≤20%. For more than 94% of pesticides, a negligible matrix effect (within ±20%) was observed. The analytical method was successfully applied and used for the detection of three urea pesticides in 4 of 11 mealworm samples.
Assuntos
Cromatografia Líquida/métodos , Resíduos de Praguicidas/análise , Praguicidas/análise , Espectrometria de Massas em Tandem/métodos , Tenebrio/efeitos dos fármacos , Acetonitrilas/química , Animais , Calibragem , Insetos Comestíveis , Hexanos/química , Insetos , Larva , Limite de Detecção , Extração em Fase Sólida , Ureia/análiseRESUMO
In this study, the levels and distribution patterns of HBCD diastereoisomers in air, water, soil, and sediment samples in South Korea were investigated after optimizing the UPLC-MS/MS analytical process. Extraction and cleanup efficiencies were tested using several different extraction solvents and adsorbents. Dichloromethane was selected as the base extraction solvent, and multi-layer silica gel (MSG) and MSG-alumina columns were selected for the removal of HBCDs from complex environmental matrices. The concentration of Æ©3 HBCDs was 22-133â¯pg/m3, 10-128â¯ng/g, 0.2-151â¯ng/L, and 0.5-552â¯ng/g dw for air, soil, water, and sediment samples, respectively. Relatively higher concentrations of Æ©3 HBCDs were observed at stations adjacent to industrial facilities (e.g., rubber and plastic, textile, chemical, fabricated metal, and wholesale trade factories) associated with the use of commercial HBCDs. The proportion of γ-HBCD in the soil (48.3-86.2%) and sediment (54.2-78.1%, except for one station) samples was similar to that found in technical and commercial HBCDs. In contrast, α-HBCD (52.3-71.2%) was dominant in all air samples, while the water samples displayed no clear trend in their diastereoisomer profiles. As the first nationwide report on HBCD diastereoisomers in the environment, this study demonstrates that most environmental compartments in South Korea are moderately contaminated with HBCDs.
Assuntos
Poluentes Atmosféricos/análise , Sedimentos Geológicos/química , Hidrocarbonetos Bromados/análise , Poluentes do Solo/análise , Poluentes Químicos da Água/análise , Poluentes Atmosféricos/química , Cromatografia Líquida , Monitoramento Ambiental/métodos , Retardadores de Chama/análise , Hidrocarbonetos Bromados/química , República da Coreia , Poluentes do Solo/química , Estereoisomerismo , Espectrometria de Massas em Tandem , Poluentes Químicos da Água/químicaRESUMO
Arsenic contamination in marine environments is a serious issue because some arsenicals are very toxic, increasing the health risks associated with the consumption of marine products. This study describes the development of an improved rapid method for the quantification of arsenic species, including arsenite (AsIII), arsenate (AsV), arsenocholine (AsC), arsenobetaine (AsB), dimethylarsinic acid (DMA), and monomethyl arsonic acid (MMA), in seaweed, sediment, and seawater samples using high-performance liquid chromatography/inductively coupled plasma-mass spectrometry (HPLC/ICP-MS). ICP-MS based on dynamic reaction cells was used to eliminate spectral interference. Ammonium nitrate- and phosphate-based eluents were used as the mobile phases for HPLC analysis, leading to shorter overall retention time (6 min) and improved peak separation. Arsenicals were extracted with a 1% HNO3 solution that required no clean-up process and exhibited reasonable sensitivity and peak resolution. The optimized method was verified by applying it to hijiki seaweed certified reference material (CRM, NMIJ 7405-a) and to spiked blank samples of sediment and seawater. The proposed method measured the concentration of AsV in the CRM as 9.6 ± 0.6 µg/kg dry weight (dw), which is close to the certified concentration (10.1 ± 0.5 µg/kg dw). The recovery of the six arsenicals was 87-113% for the sediment and 99-101% for the seawater. In the analysis of real samples, AsV was the most abundant arsenical in hijiki and gulfweed, whereas AsB was dominant in other seaweed species. The two inorganic arsenicals (AsIII and AsV) and AsV were the most dominant in the sediment and seawater samples, respectively.
Assuntos
Arsenicais/análise , Cromatografia Líquida de Alta Pressão/métodos , Monitoramento Ambiental/métodos , Espectrometria de Massas/métodos , Poluentes Químicos da Água/análise , Monitoramento Ambiental/instrumentação , Sedimentos Geológicos/química , República da Coreia , Água do Mar/química , Alga Marinha/químicaRESUMO
Interleukin-15 receptor alpha (IL-15R alpha) is a pleiotropically expressed molecule that chaperones and trans-presents IL-15 to NK and T cells. To investigate whether IL-15R alpha presented by different cells perform distinct physiological functions, we have generated four lines of mice lacking IL-15R alpha in various cell types. We find that IL-15R alpha expression on macrophages but not dendritic cells (DCs) supports the early transition of antigen specific effector CD8(+) T cells to memory cells. After memory CD8(+) T cell differentiation, IL-15R alpha expression on DCs selectively supports central memory CD8(+) T cells, whereas IL-15R alpha expression on macrophages supports both central and effector memory CD8(+) T cells. By contrast, mice lacking IL-15R alpha on macrophages, DCs, or both, exhibit equivalent defects in NK cell homeostasis and activation. These studies define unique roles for macrophage expression of IL-15R alpha and show that NK cells rely upon distinct IL-15R alpha dependent IL-15 signals than memory CD8(+) T cells. Moreover, they demonstrate the diversity, specification, and geographic restriction of cytokine signals.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Células Dendríticas/imunologia , Homeostase , Subunidade alfa de Receptor de Interleucina-15/metabolismo , Macrófagos/imunologia , Subpopulações de Linfócitos T/imunologia , Animais , Deleção de Genes , Memória Imunológica , Subunidade alfa de Receptor de Interleucina-15/genética , Células Matadoras Naturais/imunologia , Ativação Linfocitária , Camundongos , Camundongos Transgênicos , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/metabolismoRESUMO
Naturally occurring coumarins possess antibacterial and antifungal properties. In this study, these natural and synthetic coumarins were used to evaluate their antifungal activities against Aspergillus flavus, which produces aflatoxins. In addition to control antifungal activities, antiaflatoxigenic properties were also determined using a high-performance liquid chromatography in conjunction with fluorescence detection. In this study, 38 compounds tested and 4-hydroxy-7-methyl-3-phenyl coumarin showed potent antifungal and antiaflatoxigenic activities against A. flavus. Inhibitory mode of antiaflatoxigenic action by 4-hydroxy-7-methyl-3-phenyl coumarin was based on the downregulation of aflD, aflK, aflQ, and aflR in aflatoxin biosynthesis. In the cases of coumarins, antifungal and aflatoxigenic activities are highly related to the lack of diene moieties in the structures. In structurally related compounds, 2,3-dihydrobenzofuran exhibited antifungal and antiaflatoxigenic activities against A. flavus. The inhibitory mode of antiaflatoxigenic action by 2,3-dihydrobenzofuran was based on the inhibition of the transcription factor (aflS) in the aflatoxin biosynthesis pathway. These potent inhibitions of 2,3-dihydrobenzofuran and 4-hydroxy-7-methyl-3-phenyl coumarin on the Aspergillus growth and production of aflatoxins contribute to the development of new controlling agents to mitigate aflatoxin contamination.
Assuntos
Aflatoxinas/biossíntese , Antifúngicos/farmacologia , Aspergillus flavus/metabolismo , Cumarínicos/farmacologia , Aspergillus flavus/efeitos dos fármacos , Vias Biossintéticas/efeitos dos fármacos , Vias Biossintéticas/genética , Avaliação Pré-Clínica de Medicamentos , Expressão Gênica/efeitos dos fármacos , Genes FúngicosRESUMO
α-Endosulfan and some polycyclic aromatic compounds (PAHs) are persistent in the environment and can reach crop products via contaminated agricultural soils. They may even be present as mixtures in the soil and induce mixture toxicity in soil organisms such as earthworms. In this study, the combined toxicities of PAHs with α-endosulfan were determined in Eisenia fetida adults using an artificial soil system. α-Endosulfan and five PAHs were tested for their acute toxicity toward E. fetida in artificial soils. Only α-endosulfan, fluorene, and phenanthrene showed acute toxicities, with LC50 values of 9.7, 133.2, and 86.2 mg kg-1, respectively. A mixture toxicity assay was conducted using α-endosulfan at LC10 and fluorene or phenanthrene at LC50 in the artificial soils. Upon exposure to the mixture of fluorene and α-endosulfan, earthworms were killed in increasing numbers owing to their synergistic effects, while no other mixture showed any additional toxicity toward the earthworms. Along with the acute toxicity results, the biochemical and molecular changes in the fluorene- and phenanthrene-treated earthworms with or without α-endosulfan treatment demonstrated that enhancement of glutathione S-transferase activity was dependent on the addition of PAH chemicals, and the HSP70 gene expression increased with the addition of α-endosulfan. Taken together, these findings contribute toward understanding the adverse effects of pollutants when present separately or in combination with other types of chemicals.
Assuntos
Biomarcadores/análise , Endossulfano/toxicidade , Fluorenos/toxicidade , Oligoquetos/efeitos dos fármacos , Fenantrenos/toxicidade , Acetilcolinesterase/metabolismo , Animais , Carboxilesterase/metabolismo , Misturas Complexas/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Glutationa Transferase/metabolismo , Oligoquetos/fisiologia , Fosfolipídeos/metabolismo , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Poluentes do Solo/toxicidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Testes de Toxicidade AgudaRESUMO
For monitoring and risk assessment, levels and distributions of Σ29 PCBs in paddy soil samples collected from Gwangyang (10 sites) and Ulsan (20 sites), heavily industrialized cities in Korea, were investigated using high-resolution gas chromatography/high-resolution mass spectrometry. Overall, total concentrations of Σ29 PCBs in Gwangyang (216.4-978.6 pg g-1 dw) and Ulsan (273.8-1824.1 pg g-1 dw) were higher than those (106.6-222.6 pg g-1 dw) in agricultural soil from Anseong in Korea. The TEQ (toxic equivalency) values from Gwangyang (0.06-0.40 ng TEQ kg-1 dw) and Ulsan (0.06-0.22 ng TEQ kg-1 dw) were higher than those (0.04-0.11 ng TEQ kg-1 dw) in Anseong but lower than the WHO threshold level (20 ng TEQ kg-1). However, one of the most toxic congeners, PCB 126, gave the highest concentration, possibly posing a risk to the biota. Seven indicator PCB congeners contributed to 50-80% of the total concentration of Σ29 PCBs, indicating the 7 PCBs can be used as valuable indicators for monitoring. The principal component analysis and cluster analysis for the homologue profiles of PCBs indicated that all the samples from both cities had the similar PCB contamination patterns, and the major sources of the PCB contamination were most likely from the usage of Aroclor 1254 than those of Aroclors 1242 and 1260. These PCB technical mixtures were possibly significantly used by various industries including iron and steel industries in Gwangyang and petrochemical and shipbuilding industries in Ulsan.
Assuntos
Monitoramento Ambiental/métodos , Bifenilos Policlorados/análise , Medição de Risco/métodos , Poluentes do Solo/análise , Agricultura , Cidades , Cromatografia Gasosa-Espectrometria de Massas/métodos , Indústrias , Bifenilos Policlorados/toxicidade , Análise de Componente Principal , República da Coreia , Poluentes do Solo/toxicidadeRESUMO
Caffeine test solute was employed in combination with an internal standard (IS), 1,4-dimethoxybenzene, in preparative-gas chromatography (prep-GC), with nuclear magnetic resonance (NMR) experiments. The IS served to: (i) quantify the trapping efficiency of an external trapping assembly, consisting of a capillary column cryotrap at the end of the analytical column; (ii) quantify the solute response in different NMR samples; and (iii) permit correlation of expected level of response of a compound in the NMR experiment, based on relative responses of the IS and solute in the GC result. The recovery rate of caffeine from multiple injections of sample (1×, 2×, 5× and 10×) was 69.6 ± 1.3%, which correlated well (R(2) = 0.999) with the number of injections of compound. The (1)H-NMR spectrum was sufficient to enable structural characterisation of the reference caffeine compound, and was achieved with recovery of amounts of ≤10 µg from a single aliquot. Less than 400 µg of collected caffeine (40 replicate injections) was sufficient for structural characterisation by (13)C-NMR spectral analysis. The method allows development of approaches to separate unknown compounds in complex samples, and to separately use MS and NMR for their characterisation.
Assuntos
Cafeína/análise , Cromatografia Gasosa/métodos , Espectroscopia de Ressonância Magnética/métodosRESUMO
Since the introduction of the positive list system (PLS) for agricultural products in the Republic of Korea, the demand for a quick, easy multi-residue analysis method increased continuously. Herein, the quick, easy, cheap, effective, rugged, and safe (QuEChERS) technique combined with liquid chromatography−tandem mass spectrometry was employed to optimize a method for the multi-residue analysis of 287 pesticide residues in mandarin orange and grapefruit. Method validation was conducted in terms of selectivity, limit of quantitation (LOQ), linearity, accuracy, precision, and matrix effect. All the compounds at low spiking levels (1, 2.5, 5, or 10 mg/kg) could be quantified at LOQs lower than 0.01 mg/kg (PLS level). The linearity of the matrix-matched calibration curve for each compound is in the range 0.5−50 µg/L, and its coefficient of determination (R2) is >0.990. Satisfactory recovery values of 70−120% with a relative standard deviation of ≤20% are obtained for all compounds in the mandarin orange and grapefruit samples. A negligible matrix effect (−20−20%) is observed for more than 94.8% and 85.4% of the pesticides in mandarin orange and grapefruit, respectively. Therefore, this analytical method can contribute to pesticide residue analyses of citrus fruits for routine laboratory testing.
RESUMO
BACKGROUND: A growing body of evidence suggests that T-cell responses against neoantigens are critical regulators of response to immune checkpoint blockade. We previously showed that circulating neoantigen-specific CD8 T cells in patients with lung cancer responding to anti-Programmed death-ligand 1 (PD-L1) (atezolizumab) exhibit a unique phenotype with high expression of CD57, CD244, and KLRG1. Here, we extended our analysis on neoantigen-specific CD8 T cells to patients with metastatic urothelial cancer (mUC) and further profiled total CD8 T cells to identify blood-based predictive biomarkers of response to atezolizumab. METHODS: We identified tumor neoantigens from 20 patients with mUC and profiled their peripheral CD8 T cells using highly multiplexed combinatorial tetramer staining. Another set of patients with mUC treated with atezolizumab (n=30) or chemotherapy (n=40) were selected to profile peripheral CD8 T cells by mass cytometry. Using single-cell transcriptional analysis (single-cell RNA sequencing (scRNA-seq)), together with CITE-seq (cellular indexing of transcriptomes and epitopes by sequencing) and paired T-cell receptor (TCR) sequencing, we further characterized peripheral CD8 T cells in a subset of patients (n=16). RESULTS: High frequency of CD57 was observed in neoantigen-specific CD8 T cells in patients with mUC responding to atezolizumab. Extending these findings to bulk CD8 T cells, we found higher frequency of CD57 expressing CD8 T cells before treatment in patients responding to atezolizumab (n=20, p<0.01) but not to chemotherapy. These findings were corroborated in a validation cohort (n=30, p<0.01) and notably were independent of known biomarkers of response. scRNA-seq analysis identified a clonally expanded cluster enriched within CD57+ CD8 T cells in responding patients characterized by higher expression of genes associated with activation, cytotoxicity, and tissue-resident memory markers. Furthermore, compared with CD57- CD8 T cells, TCRs of CD57+ CD8 T cells showed increased overlap with the TCR repertoire of tumor-infiltrating T cells. CONCLUSIONS: Collectively, we show high frequencies of CD57 among neoantigen-specific and bulk CD8 T cells in patients responding to atezolizumab. The TCR repertoire overlap between peripheral CD57+ CD8 T cells and tumor-infiltrating lymphocytes suggest that accumulation of peripheral CD57+ CD8 T cells is reflective of an ongoing antitumor T-cell response. Our findings provide evidence and rationale for using circulating CD8 T cells expressing CD57 as a readily accessible blood-based biomarker for selecting patients with mUC for atezolizumab therapy.
Assuntos
Carcinoma de Células de Transição , Neoplasias Pulmonares , Antígeno B7-H1/metabolismo , Antígenos CD57/imunologia , Linfócitos T CD8-Positivos , Humanos , Receptores de Antígenos de Linfócitos T , Análise de Célula ÚnicaRESUMO
Traditionally in Korea, Protaetia brevitarsis seulensis (white-spotted flower chafer) has been used as a medicine, and recently has attracted increased attention due to its antithrombotic efficacy. Some of spent mushroom compost or fermented oak sawdust, a feedstock for P. brevitarsis, were contaminated with three fungicides, carbendazim, dimethomorph, and fenoxanil, which could be transferred to the insect. This study was aimed to optimize a simple extraction method combined with liquid chromatography tandem mass spectrometry and apply it to the real samples. After the pulverized samples (5 g) were extracted with acetonitrile (10 mL) and formic acid (100 µL), fat and lipids in the samples were slowly precipitated at -20°C for 24 hours. After eight different clean-up methods were investigated, the mixture of 150 mg MgSO4/25 mg PSA/25 mg C18 was selected due to optimal recovery of the target compounds. Recovery (77.9%â80.8% for carbendazim, 111.2%â116.7% for dimethomorph, and 111.9%â112.5% for fenoxanil) was achieved with reasonable relative standard deviation (<5.5%) The analytical method developed in this study was used to analyze three compounds in the 24 insect samples donated by the insect farm owners but no target compounds were detected. These results can provide important data for establishing the pesticide safety standards for P. brevitarsis before the medical applications.
Assuntos
Benzimidazóis/análise , Carbamatos/análise , Besouros/química , Morfolinas/análise , Espectrometria de Massas em Tandem , Acetonitrilas/química , Animais , Cromatografia Líquida , Reprodutibilidade dos TestesRESUMO
Tenebrio molitor larva (mealworms) has recently attracted attention as a protein source for food and feed. The larva is generally fed with wheat bran, which can be possibly contaminated with glyphosate. To establish food safe standards, a rugged and effective analytical method for glyphosate, aminomethylphosphonic acid, glufosinate, and their metabolites including 3-methylphosphinico-propionic acid, and N-acetyl glufosinate, in mealworms was optimized using liquid chromatography tandem mass spectrometry. An anionic polar pesticide column was used due to its high suitability for glyphosate. Acidified water and acetonitrile were used to extract the target compounds without contribution from various fatty and pigment interferences derived from brownish insects. Seven different clean-up procedures ((1) 50 mg C18 (2) 20 mg C18/Z-sep (3) PRiME hydrophilic-lipophilic balance (HLB) cartridge (4) 75 mg Z-sep, (5) 75 mg Z-sep+, (6) EMR-lipid cartridge, and (7) 50 mg ENVI-Carb) were compared. Due to its simplicity and cost-effectiveness, PRiME HLB was selected for clean-up. The recoveries of the target compounds were ranged from 86 to 96% with < 20% relative standard deviations. Therefore, this simple and effective method can be applied for the two pesticides and their metabolites in other edible insects or high-fat matrices.
Assuntos
Aminobutiratos/análise , Glicina/análogos & derivados , Herbicidas/análise , Larva/metabolismo , Tenebrio/metabolismo , Animais , Cromatografia Líquida/métodos , Glicina/análise , Espectrometria de Massas em Tandem/métodos , GlifosatoRESUMO
We investigated the levels and distribution patterns of α- and ß-endosulfan and endosulfan sulfate in air, soil, water, and sediment samples collected from the South Korean persistent organic pollutants (POPs) monitoring networks. In the air samples, the highest concentrations of the total (Σ3) endosulfan (50.3-611 pg/m3, mean: 274 pg/m3) were observed during summer. Spearman analysis revealed a good correlation between agricultural land area and atmospheric concentrations of Σ3 endosulfan except during winter. Regardless of the season, the ratio of the two isomers (α/ß) was 3.6-4.9 in the air samples, higher than that observed in technical mixtures (2.0-2.3), possibly due to the higher volatility of α-endosulfan, compared to ß-endosulfan. Concentrations of Σ3 endosulfan in the soil samples (n.d.-13.4 ng/g, mean: 0.8 ng/g) were not significantly different except at some stations adjacent to large areas of farmland. The average levels of Σ3 endosulfan in the water and sediment samples were 2.1 ng/L and 0.1 ng/g dw, respectively. In analyzing the four largest rivers, it was observed that a few water stations during spring and fall and sediment stations in fall had high concentrations of the two isomers and endosulfan sulfate, particularly around the Yeoungsan and Nakdong Rivers near large areas of agricultural land. Endosulfan sulfate was dominant at most water and sediment sampling stations. This study demonstrates that the endosulfan found in most environmental compartments most probably derives from agricultural areas despite its ban as a pesticide. On the other hand, given that it was also detected in industrial and urban areas, in which pesticide application does not occur, it can be conjectured that endosulfan is aerially transported at higher temperatures and continuously circulates within the environment.
Assuntos
Endossulfano/análise , Inseticidas/análise , Monitoramento Ambiental , República da Coreia , Solo , ÁguaRESUMO
BACKGROUND: Noroviruses are the leading cause of epidemic acute gastroenteritis and foodborne diarrheal disease in humans. However, there are no approved vaccines for noroviruses. Potential correlates of protection identified through human challenge studies include mucosal IgA, memory B cells, and serum-blocking antibody titers (BT50). METHODS: We conducted a single-site, randomized, double-blind, placebo-controlled clinical trial of an oral norovirus vaccine to determine safety and immunogenicity. This tablet vaccine is comprised of a nonreplicating adenovirus-based vector expressing the VP1 gene from the GI.1 norovirus strain and a double-stranded RNA adjuvant. Sixty-six adult subjects meeting inclusion/exclusion criteria were randomized 2:1 to receive a single vaccine dose or placebo, respectively. Immunogenicity was primarily assessed by serum BT50. Additional outcomes included serum ELISA titers, fecal and saliva antibody titers, memory and antibody-secreting cell (ASC) frequency, and B cell phenotyping. RESULTS: The vaccine was well-tolerated, with no dose-limiting toxicities. Adverse events were mild or moderate. The primary immunological endpoint (increase in BT50 titers) was met in the high-dose group (P = 0.0003), with 78% showing a ≥2-fold rise in titers after a single immunization. Vaccine recipients also developed mucosally primed VP1-specific circulating ASCs, IgA+ memory B cells expressing gut-homing receptor (α4ß7), and fecal IgA, indicating substantial and local responses potentially relevant to prevent norovirus infection. CONCLUSION: This oral norovirus vaccine was well-tolerated and generated substantial immune responses, including systemic and mucosal antibodies as well as memory IgA/IgG. These results are a major step forward for the development of a safe and immunogenic oral norovirus vaccine. TRIAL REGISTRATION: ClinicalTrials.gov NCT02868073. FUNDING: Vaxart.
Assuntos
Administração Oral , Infecções por Caliciviridae/prevenção & controle , Norovirus , Comprimidos/administração & dosagem , Comprimidos/farmacologia , Vacinas Virais/administração & dosagem , Vacinas Virais/efeitos adversos , Vacinas Virais/imunologia , Imunidade Adaptativa , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Linfócitos B , Infecções por Caliciviridae/virologia , Método Duplo-Cego , Doenças Transmitidas por Alimentos/prevenção & controle , Doenças Transmitidas por Alimentos/virologia , Gastroenterite/prevenção & controle , Humanos , Imunoglobulina A , Norovirus/genética , Estados Unidos , Proteínas Estruturais Virais/genéticaRESUMO
A multiresidue method for the simultaneous and rapid analysis of 360 pesticides in representative agricultural produce (brown rice, orange, spinach, and potato) was developed using a modified QuEChERS procedure combined with gas chromatography-tandem mass spectrometry (GC-MS/MS). Selected reaction monitoring transition parameters (e.g., collision energy, precursor and product ions) in MS/MS were optimized to achieve the best selectivity and sensitivity for a wide range of GC-amenable pesticides. A short (20 m) microbore (0.18 mm i.d.) column resulted in better signal-to-noise ratio with reduced analysis time than a conventional narrowbore column (30 m × 0.25 mm i.d.). The priming injection dramatically increased peak areas by masking effect on a new GC liner. The limit of quantitation was <0.01 mg/kg, and the correlation coefficients (r2) of matrix-matched standards were >0.99 within the range of 0.0025-0.1 mg/kg. Acetonitrile with 0.1% formic acid without additional buffer salts was used for pesticide extraction, whereas only primary-secondary amine (PSA) was used for dispersive solid phase extraction (dSPE) cleanup, to achieve good recoveries for most of the target analytes. The recoveries ranged from 70 to 120% with relative standard deviations of ≤20% at 0.01 and 0.05 mg/kg spiking levels (n = 6) in all samples, indicating acceptable accuracy and precision of the method. Seventeen real samples from local markets were analyzed by using the optimized method, and 14 pesticides in 11 incurred samples were found at below the maximum residue limits.
Assuntos
Citrus sinensis/química , Contaminação de Alimentos/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Oryza/química , Resíduos de Praguicidas/química , Solanum tuberosum/química , Spinacia oleracea/química , Limite de DetecçãoRESUMO
Plant essential oils from 40 plant species were tested for their insecticidal activities against larvae of Lycoriella ingénue (Dufour) using a fumigation bioassay. Good insecticidal activity against larvae of L. ingenua was achieved with essential oils of Chenopodium ambrosioides L., Eucalyptus globulus Labill, Eucalyptus smithii RT Baker, horseradish, anise and garlic at 10 and 5 microL L(-1) air. Horseradish, anise and garlic oils showed the most potent insecticidal activities among the plant essential oils. At 1.25 microL L(-1), horseradish, anise and garlic oils caused 100, 93.3 and 13.3% mortality, but at 0.625 microL L(-1) air this decreased to 3.3, 0 and 0% respectively. Analysis by gas chromatography-mass spectrometry led to the identification of one major compound from horseradish, and three each from anise and garlic oils. These seven compounds and m-anisaldehyde and o-anisaldehyde, two positional isomers of p-anisaldehyde, were tested individually for their insecticidal activities against larvae of L. ingenua. Allyl isothiocyanate was the most toxic, followed by trans-anethole, diallyl disulfide and p-anisaldehyde with LC(50) values of 0.15, 0.20, 0.87 and 1.47 microL L(-1) respectively.
Assuntos
Armoracia/química , Dípteros , Alho/química , Inseticidas/análise , Pimpinella/química , Animais , Larva , Óleos Voláteis/química , Plantas/químicaRESUMO
There are several benefits of oral immunization including the ability to elicit mucosal immune responses that may protect against pathogens that invade through a mucosal surface. Our understanding of human immune biology is hampered by the difficulty in isolating mucosal cells from humans, and the fact that animal models may or may not completely mirror human intestinal immunobiology. In this human pharmacodynamic study, a novel adenovirus vector-based platform expressing influenza hemagglutinin was explored. We used radio-controlled capsules to deliver the vaccine to either the jejunum or the ileum. The resulting immune responses induced by immunization at each of the intestinal sites were investigated. Both intestinal sites were capable of inducing mucosal and systemic immune responses to influenza hemagglutinin, but ileum delivery induced higher numbers of antibody secreting cells of IgG and IgA isotypes, increased mucosal homing B cells, and higher number of vaccine responders. Overall, these data provided substantial insights into human mucosal inductive sites, and aided in the design and selection of indications that could be used with this oral vaccine platform.
Assuntos
Anticorpos Antivirais/sangue , Vacinas contra Influenza/administração & dosagem , Influenza Humana/prevenção & controle , Mucosa Intestinal/imunologia , Vacinação , Adenoviridae/genética , Administração Oral , Adolescente , Adulto , Animais , Anticorpos Neutralizantes/sangue , Cães , Vetores Genéticos , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Humanos , Vacinas contra Influenza/imunologia , Influenza Humana/sangue , Influenza Humana/imunologia , Mucosa Intestinal/metabolismo , Leucócitos/imunologia , Células Madin Darby de Rim Canino , Pessoa de Meia-Idade , Potência de Vacina , Tecnologia sem Fio , Adulto JovemRESUMO
The concentrations, profiles, and source-receptor relationships of seven indicator polychlorinated biphenyls (PCBs) (#28, 52, 101, 118, 138, 153, and 180) found in soil at 25 rural, urban, and industrial sites in Ulsan, South Korea were investigated. For this study, 75 soil samples were collected, 25 each in January of 2011, 2012, and 2013. Principal component analysis was used to evaluate the influence of the emission sources on the soil samples. The concentrations of total seven PCBs (Σ7 PCBs) ranged between 0.034 ng/g and 143 ng/g (mean: 5.10 ng/g, median: 0.440 ng/g), which indicated slight or moderate contamination levels, respectively, compared to those in the other countries or other cities in Korea. The concentrations of Σ7 PCBs at the industrial and urban sites were significantly higher than those at the rural sites, due to the direct influence of emission sources related to industrial activities rather than urban emission sources. Generally, the profiles of PCBs were dominated by penta- and hexa-chlorinated biphenyls at all the study sites, suggesting common sources of PCBs in Ulsan. PCB source identification indicated that leakage from transformer oils in the major industrial complexes and PCB-containing paints used in the automobile and shipbuilding industrial complexes were possibly the main sources of indicator PCBs in the study areas.