Aquatic macrophyte growth season in Central and Northern European Union and the implications for aquatic macrophyte risk assessments for herbicides.

Under current European Union regulation, the risks to aquatic organisms must be assessed for uses of plant protection products (PPPs) that may result in exposure to the environment. For herbicidal PPPs, aquatic macrophytes are often the most sensitive taxa. For some herbicidal modes of action, macrophytes may be affected only while they are actively growing. For the risk assessment, it is therefore useful to know whether application timings would result in surface water exposure during periods when aquatic macrophytes are actively growing (therefore potentially resulting in effects). Toxicity endpoints, which are based on studies with active growth, may be overconservative in cases where exposure of PPPs will not co-occur with active macrophyte growth. A comprehensive literature search was performed, using systematic and manual approaches, with the aim of identifying the main active growth period for macrophytes in natural freshwater bodies in climates relevant to the Central and Northern zones of the European Union. The results of the searches were screened initially to identify all potentially relevant references, for which a full evaluation was then performed. Reliability was assessed using the principles of the Klimisch scoring system. As part of the full evaluation, growth periods were identified for each macrophyte species studied. Finally, the extracted growth periods were considered together to determine an overall active growth period for aquatic macrophytes representative of the Central and Northern EU zones. Based on this literature review, the active growth period identified for most aquatic macrophyte species representative of the Central and Northern EU zones is April to September. Relating to the regulatory implication of these results, it may be possible to conclude a low risk for aquatic macrophytes if the predicted surface water exposure period for certain PPPs is demonstrated to be outside the periods of active growth. Integr Environ Assess Manag 2023;00:1-15. © 2023 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Efficacy of ceftazidime in a murine model following a lethal aerosol exposure to Burkholderia pseudomallei.

Melioidosis is an endemic disease in numerous tropical regions. Additionally, the bacterium that causes melioidosis, Burkholderia pseudomallei, has potential to be used as a biological weapon. Therefore, development of effective and affordable medical countermeasures to serve regions affected by the disease and to have medical countermeasures available in the event of a bioterrorism attack remains critical. The current study evaluated the efficacy of eight distinct acute phase ceftazidime treatment regimens administered therapeutically in the murine model. At the conclusion of the treatment period, survival rates were significantly greater in several of the treated groups when compared to the control group. Pharmacokinetics of a single dose of ceftazidime were examined at 150 mg/kg, 300 mg/kg, and 600 mg/kg and were compared to an intravenous clinical dose administered at 2000 mg every eight hours. The clinical dose has an estimated 100% fT > 4*MIC which exceeded the highest murine dose of 300 mg/kg every six hours at 87.2% fT > 4*MIC. Based upon survival at the end of the treatment regimen and supplemented by pharmacokinetic modeling, a daily dose of 1200 mg/kg of ceftazidime, administered every 6 h at 300 mg/kg, provides protection in the acute phase of inhalation melioidosis in the murine model.

Neutralizing Concentrations of Anti-Botulinum Toxin Antibodies Positively Correlate with Mouse Neutralization Assay Results in a Guinea Pig Model.

Botulinum neurotoxins (BoNT) are some of the most toxic proteins known and can induce respiratory failure requiring long-term intensive care. Treatment of botulism includes the administration of antitoxins. Monoclonal antibodies (mAbs) hold considerable promise as BoNT therapeutics and prophylactics, due to their potency and safety. A three-mAb combination has been developed that specifically neutralizes BoNT serotype A (BoNT/A), and a separate three mAb combination has been developed that specifically neutralizes BoNT serotype B (BoNT/B). A six mAb cocktail, designated G03-52-01, has been developed that combines the anti-BoNT/A and anti-BoNT/B mAbs. The pharmacokinetics and neutralizing antibody concentration (NAC) of G03-52-01 has been determined in guinea pigs, and these parameters were correlated with protection against an inhalation challenge of BoNT/A1 or BoNT/B1. Previously, it was shown that each antibody demonstrated a dose-dependent mAb serum concentration and reached maximum circulating concentrations within 48 h after intramuscular (IM) or intraperitoneal (IP) injection and that a single IM injection of G03-52-01 administered 48 h pre-exposure protected guinea pigs against an inhalation challenge of up to 93 LD50s of BoNT/A1 and 116 LD50s of BoNT/B1. The data presented here advance our understanding of the relationship of the neutralizing NAC to the measured circulating antibody concentration and provide additional support that a single IM or intravenous (IV) administration of G03-52-01 will provide pre-exposure prophylaxis against botulism from an aerosol exposure of BoNT/A and BoNT/B.

Toxicokinetic evaluation of the common indoor air pollutant, α-pinene, and its potential reactive metabolite, α-pinene oxide, following inhalation exposure in rodents.

The toxicokinetic behavior of α-pinene and its potential reactive metabolite, α-pinene oxide, was investigated following whole body inhalation exposure to 50 and 100 ppm α-pinene in rats and mice for 6 h per day for 7d. In both species and sexes, the maximum blood concentration (Cmax) increased more than proportionally while the increase in area under the concentration time curve (AUC) was proportional to the exposure concentration. When normalized to the calculated dose (D), both Cmax/D (male rats, 12.2-54.5; female rats, 17.4-74.1; male mice, 7.41-14.2; female mice, 6.59-13.0 (ng/mL)/(mg/kg)) and AUC/D (male rats, 28.9-31.1; female rats, 55.8-56.8; male mice, 18.1-19.4; female mice, 19.2-22.5 (h*ng/mL)/(mg/kg)) in rats were higher than in mice and in female rats were higher than in male rats; no sex difference was observed in mice. α-Pinene was eliminated from blood with half-lives between 12.2 and 17.4 h in rats and 6.18-19.4 h in mice. At the low dose, the ratio of α-pinene oxide to α-pinene, based on Cmax and AUC, respectively, was 0.200-0.237 and 0.279-0.615 in rats and 0.060-0.086 and 0.036-0.011 in mice demonstrating lower formation of the oxide in mice than in rats. At the high dose, the ratio decreased considerably in both species pointing to saturation of pathways leading to the formation of α-pinene oxide. α-Pinene and the oxide were quantified in the mammary glands of rats and mice with tissue to blood ratios of ≥23 demonstrating retention of these analytes in mammary glands. The findings of epoxide formation and species- and sex-differences in systemic exposure may be important in providing context and relating animal findings to human exposures.

In Vitro and In Vivo Characterization of Tebipenem (TBP), an Orally Active Carbapenem, against Biothreat Pathogens.

Bacillus anthracis and Yersinia pestis, causative pathogens for anthrax and plague, respectively, along with Burkholderia mallei and B. pseudomallei are potential bioterrorism threats. Tebipenem pivoxil hydrobromide (TBP HBr, formerly SPR994), is an orally available prodrug of tebipenem, a carbapenem with activity versus multidrug-resistant (MDR) gram-negative pathogens, including quinolone-resistant and extended-spectrum-ß-lactamase-producing Enterobacterales. We evaluated the in vitro activity and in vivo efficacy of tebipenem against biothreat pathogens. Tebipenem was active in vitro against 30-strain diversity sets of B. anthracis, Y. pestis, B. mallei, and B. pseudomallei with minimum inhibitory concentration (MIC) values of 0.001 - 0.008 µg/ml for B. anthracis, ≤0.0005 - 0.03 µg/ml for Y. pestis, 0.25 - 1 µg/ml for B. mallei, and 1 - 4 µg/ml for B. pseudomallei In a B. anthracis murine model, all control animals died within 52 h post challenge. The survival rates in the groups treated with tebipenem were 75% and 73% when dosed at 12 h and 24 h post challenge, respectively. The survival rates in the positive control groups treated with ciprofloxacin were 75% and when dosed 12 h and 25% when dosed 24 h post challenge, respectively. Survival rates were significantly (p=0.0009) greater in tebipenem groups treated at 12 h and 24 h post challenge and in the ciprofloxacin group 12 h post-challenge vs. the vehicle-control group. For Y. pestis, survival rates for all animals in the tebipenem and ciprofloxacin groups were significantly (p<0.0001) greater than the vehicle-control group. These results support further development of tebipenem for treating biothreat pathogens.

An Electronic Clinical Decision-Making Tool for Patients with Suspected Colorectal Cancer-Preliminary Evaluation in Patients Presenting with Rectal Bleeding.

BACKGROUND AND OBJECTIVES: The CarePath-CRC electronic clinical decision-making application was designed to assist physicians with evaluation of patients with suspected colorectal cancer (CRC). The physician completes an interactive checklist of evidence-based clinical parameters, and a recommended referral urgency is generated based on the post-test probability of CRC. This study aimed toward validation of the tool in symptomatic patients presenting with rectal bleeding. METHODS: The medical records of a sample of patients with histologically confirmed CRC from 2010 to 2014 were reviewed. The CarePath-CRC tool was applied retrospectively to all patients who initially presented with rectal bleeding, to determine its sensitivity for detecting CRC in this population. A generated recommendation of 'immediate referral' (referral ≤24 hours, expected endoscopy ≤2 weeks) or 'urgent referral' (expected consultation and endoscopy ≤4 and ≤8 weeks) was considered a positive test result. An a priori sensitivity of 90% was deemed adequate, based on test characteristics of the tool's individual clinical criteria. RESULTS: The tool was applied to 281 patients. A total of 69 (24.6%) and 211 (75.1%) patients met criteria for immediate and urgent referral, respectively. The remaining patient (0.4%) met criteria for 'possible priority referral', while none met criteria for 'no specific action recommended'. This resulted in a calculated sensitivity of 99.6% (95% confidence interval 98.0 to 99.9%). CONCLUSIONS: The CarePath-CRC tool is sensitive in the prediction of CRC in patients presenting with rectal bleeding. A prospective cohort study is being designed to allow for acquisition of comprehensive test performance characteristics and full validation of the instrument.

Metabolic Basis for Nonlinearity in 1,3-Dichloropropene Toxicokinetics and Use in Setting a Kinetically-derived Maximum Inhalation Exposure Concentration in Mice.

1,3-Dichloropropene (1,3-D) showed a statistically increased incidence of bronchioloalveolar adenomas in male B6C3F1 mice at 60 ppm air concentration during previous chronic inhalation testing. No tumors were observed in female mice, nor in either sex of F344 rats up to 60 ppm, the highest dose tested. Therefore, to understand if lung tumors observed in high dose male mice are due to saturation of metabolic clearance, the linearity of 1,3-D concentrations in mouse blood was investigated on day 15 of repeated nose-only inhalation exposure to 0, 10, 20, 40, 60, 90, and 120 ppm (6 h/d, 7 d/week). Additional groups were included at 20, 60, and 120 ppm for blood collection at 1.5 and 3 h of exposure and up to 25 or 40 min post-exposure to determine area-under-the-curve. The data provide multiple lines of evidence that systemic exposures to 1,3-D in the mouse become nonlinear at inhalation exposure levels of 30 ppm or above. A reduction in minute volume occurred at the highest exposure concentration. The glutathione (GSH)-dependent metabolism of 1,3-D results in significant depletion of GSH at repeated exposure levels of 30 ppm and above. This loss of GSH results in decreased metabolic clearance of this test material, with a concomitant increase of the 1,3-D isomers in circulating blood at exposure concentrations ≥30 ppm. Shifts in the ratio of cis- and trans-1,3-D also support nonlinear toxicokinetics well below 60 ppm. Based on this data, a kinetically derived maximum dose for 1,3-D in mice for repeated exposures should be at or below 30 ppm. These results support non-relevance of 1,3-D-induced benign pulmonary tumorigenicity in mice for human health risk assessment.

A Factorial Analysis of Drug and Bleeding Effects in Toxicokinetic Studies.

The ICH revised the S3A guidance allowing blood to be microsampled for toxicokinetic analysis from the main study cohorts of rats in general toxicology studies. The resulting changes in the hemogram have been examined in healthy animals but the ability to read through the data when there are toxicological changes has not been thoroughly examined in the literature. To address this, a toxicology study in Sprague Dawley rats was conducted where animals received repeated doses of saline or valproic acid by IP injection daily for 7 days. Animals in both treatment groups were unbled, serially bled (6 bleeds/animal at 0.1 ml/bleed) or compositely bled (2 bleeds/animal at 0.6 ml/bleed) on days 1 and 7 for TK analysis. No statistically significant changes in the clinical pathology were observed for either the serial bleed or composite bleed animals when compared with their respective unbled control; however, a 4%-7% decrease in erythrocyte counts following serial bleeding and a 5%-19% decrease following composite bleeding was observed. When all the clinical pathology and organ weight data were equivalence tested, both the serial bleed and composite bleed results were equivalent to their unbled controls except for the erythroid parameters in the composite bleed group. Toxicokinetic analysis of the blood samples resulted in comparable concentration-time curves, regardless of the method of blood collection. Under these study conditions, the results show blood microsamples can be collected from the core or recovery cohort of animals in a toxicology study without impacting the toxicological interpretation in rats.

Iron oxide nanoclusters for T 1 magnetic resonance imaging of non-human primates.

Iron-oxide-based contrast agents for magnetic resonance imaging (MRI) had been clinically approved in the United States and Europe, yet most of these nanoparticle products were discontinued owing to failures to meet rigorous clinical requirements. Significant advances have been made in the synthesis of magnetic nanoparticles and their biomedical applications, but several major challenges remain for their clinical translation, in particular large-scale and reproducible synthesis, systematic toxicity assessment, and their preclinical evaluation in MRI of large animals. Here, we report the results of a toxicity study of iron oxide nanoclusters of uniform size in large animal models, including beagle dogs and the more clinically relevant macaques. We also show that iron oxide nanoclusters can be used as T 1 MRI contrast agents for high-resolution magnetic resonance angiography in beagle dogs and macaques, and that dynamic MRI enables the detection of cerebral ischaemia in these large animals. Iron oxide nanoclusters show clinical potential as next-generation MRI contrast agents.

Molecular-matched materials for anticancer drug delivery and imaging.

AIM: In this study, we aim to construct nanoformulation with high-cargo loading and controlled serum kinetics. MATERIALS & METHODS: Molecular-matched materials (MMMs) are established through the conjugation of the functional moiety to a molecule representative of the nanoparticle's core. Molecular-matched nanoemulsions and liposomes were prepared using MMMs. RESULTS: This technique based on MMMs even allows us to efficiently load either hydrophobic or hydrophilic moieties into a hydrophobic core of the nanoparticles. MMMs-based nanoparticles showed marked improvement in serum pharmacokinetics and anticancer effect. CONCLUSION: The desired performance can be achieved when the hydrophobic anchor of the PEG derivatives and the moiety conjugated to the therapeutic (or imaging) agents are molecularly identical to the core.

pH-sensitive nanoformulated triptolide as a targeted therapeutic strategy for hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) has one of the worst prognoses for survival as it is poorly responsive to both conventional chemotherapy and mechanism-directed therapy. This results from a lack of therapeutic concentration in the tumor tissue coupled with the highly toxic off-site effects exhibited by these compounds. Consequently, we believe the best packaging for holistic therapy for HCC will involve three components: a potent therapeutic, a rationally designed drug delivery vehicle to enrich the target site concentration of the drug, and a surface ligand that can enable a greater propensity to internalization by tumor cells compared to the parenchyma. We screened a library containing hundreds of compounds against a panel of HCC cells and found the natural product, triptolide, to be more effective than sorafenib, doxorubicin, and daunorubicin, which are the current standards of therapy. However, the potential clinical application of triptolide is limited due to its poor solubility and high toxicity. Consequently, we synthesized tumor pH-sensitive nanoformulated triptolide coated with folate for use in an HCC-subpopulation that overexpresses the folate receptor. Our results show triptolide itself can prevent disease progression, but at the cost of significant toxicity. Conversely, our pH-sensitive nanoformulated triptolide facilitates uptake into the tumor, and specifically tumor cells, leading to a further increase in efficacy while mitigating systemic toxicity.

A general strategy for site-directed enzyme immobilization by using NiO nanoparticle decorated mesoporous silica.

Mesoporous materials have recently gained much attention owing to their large surface area, narrow pore size distribution, and superior pore structure. These materials have been demonstrated as excellent solid supports for immobilization of a variety of proteins and enzymes for their potential applications as biocatalysts in the chemical and pharmaceutical industries. However, the lack of efficient and reproducible methods for immobilization has limited the activity and recyclability of these biocatalysts. Furthermore, the biocatalysts are usually not robust owing to their rapid denaturation in bulk solvents. To solve these problems, we designed a novel hybrid material system, mesoporous silica immobilized with NiO nanoparticles (SBA-NiO), wherein enzyme immobilization is directed to specific sites on the pore surface of the material. This yielded the biocatalytic species with higher activity than free enzyme in solution. These biocatalytic species are recyclable with minimal loss of activity after several cycles, demonstrating an advantage over free enzymes.

Multifunctional tumor pH-sensitive self-assembled nanoparticles for bimodal imaging and treatment of resistant heterogeneous tumors.

Nanoparticle-based diagnosis-therapy integrative systems represent an emerging approach to cancer treatment. However, the diagnostic sensitivity, treatment efficacy, and bioavailability of nanoparticles as well as the heterogeneity and drug resistance of tumors pose tremendous challenges for clinical implementation. We herein report on the fabrication of tumor pH-sensitive magnetic nanogrenades (termed PMNs) composed of self-assembled iron oxide nanoparticles and pH-responsive ligands. These PMNs can readily target tumors via surface-charge switching triggered by the acidic tumor microenvironment, and are further disassembled into a highly active state in acidic subcellular compartments that "turns on" MR contrast, fluorescence and photodynamic therapeutic activity. We successfully visualized small tumors implanted in mice via unique pH-responsive T1MR contrast and fluorescence, demonstrating early stage diagnosis of tumors without using any targeting agents. Furthermore, pH-triggered generation of singlet oxygen enabled pH-dependent photodynamic therapy to selectively kill cancer cells. In particular, we demonstrated the superior therapeutic efficacy of PMNs in highly heterogeneous drug-resistant tumors, showing a great potential for clinical applications.

Fatty acids as therapeutic auxiliaries for oral and parenteral formulations.

Many drugs have decreased therapeutic activity due to issues with absorption, distribution, metabolism and excretion. The co-formulation or covalent attachment of drugs with fatty acids has demonstrated some capacity to overcome these issues by improving intestinal permeability, slowing clearance and binding serum proteins for selective tissue uptake and metabolism. For orally administered drugs, albeit at low level of availability, the presence of fatty acids and triglycerides in the intestinal lumen may promote intestinal uptake of small hydrophilic molecules. Small lipophilic drugs or acylated hydrophilic drugs also show increased lymphatic uptake and enhanced passive diffusional uptake. Fatty acid conjugation of small and large proteins or peptides has exhibited protracted plasma half-lives, site-specific delivery and sustained release upon parenteral administration. These improvements are most likely due to associations with lipid-binding serum proteins, namely albumin, LDL and HDL. These molecular interactions, although not fully characterized, could provide the ability of using the endogenous carrier systems for improving therapeutic outcomes.

A dicarboxylic fatty acid derivative of paclitaxel for albumin-assisted drug delivery.

Paclitaxel (PTX) is a potent chemotherapy for many cancers but it suffers from very poor solubility. Consequently, the TAXOL formulation uses copious amounts of the surfactant Cremophor EL to solubilize the drug for injection, resulting in severe hypersensitivity and neutropenia. In contrast to Cremophor EL, presented is a way to solubilize PTX by conjugation of a dicarboxylic fatty acid for specific binding to the ubiquitous protein, serum albumin. The conjugation chemistry was simplified to a single step using the activated anhydride form of 3-pentadecylglutaric (PDG) acid, which is reactive to a variety of nucleophiles. The PDG derivative is less cytotoxic than the parent compound and was found to slowly hydrolyze to PTX (≈ 5% over 72 h) in serum, tumor cytosol, and tumor tissue homogenate. When injected intravenously to tumor-bearing mice, [(3) H]-PTX in the TAXOL formulation was cleared rapidly with a half-life of 7 h. In the case of the PDG derivative of PTX, the drug is quickly distributed and approximately 20% of the injected dose remained in the vasculature experiencing a 23 h half-life. These improvements from modifying PTX with the PDG fatty acid present the opportunity for PDG to become a generic modification for the improvement of many therapeutics.

Tumor-targeted delivery of siRNA by self-assembled nanoparticles.

We have developed a self-assembled nanoparticle (NP) that efficiently delivers small interfering RNA (siRNA) to the tumor by intravenous (IV) administration. The NP was obtained by mixing carrier DNA, siRNA, protamine, and lipids, followed by post-modification with polyethylene glycol and a ligand, anisamide. Four hours after IV injection of the formulation into a xenograft model, 70-80% of injected siRNA/g accumulated in the tumor, approximately 10% was detected in the liver and approximately 20% recovered in the lung. Confocal microscopy showed that fluorescent-labeled siRNA was efficiently delivered into the cytoplasm of the sigma receptor expressing NCI-H460 xenograft tumor by the targeted NPs, whereas free siRNA and non-targeted NPs showed little uptake. Three daily injections (1.2 mg/kg) of siRNA formulated in the targeted NPs silenced the epidermal growth factor receptor (EGFR) in the tumor and induced approximately 15% tumor cell apoptosis. Forty percent tumor growth inhibition was achieved by treatment with targeted NPs, while complete inhibition lasted for 1 week when combined with cisplatin. The serum level of liver enzymes and body weight monitoring during the treatment indicated a low level of toxicity of the formulation. The carrier itself also showed little immunotoxicity (IMT).

133Xe contamination found in internal bacteria filter of xenon ventilation system.

OBJECTIVE: We report on (133)Xe contamination found in the reusable internal bacteria filter of our xenon ventilation system. METHODS: Internal bacteria filters (n = 6) were evaluated after approximately 1 mo of normal use. The ventilation system was evacuated twice to eliminate (133)Xe in the system before removal of the filter. Upon removal, the filter was monitored using a survey meter with an energy-compensated probe and was imaged on a scintillation camera. The filter was monitored and imaged over several days and was stored in a fume hood. RESULTS: Estimated (133)Xe activity in each filter immediately after removal ranged from 132 to 2,035 kBq (3.6-55.0 micro Ci), based on imaging. Initial surface radiation levels ranged from 0.4 to 4.5 micro Sv/h (0.04-0.45 mrem/h). The (133)Xe activity did not readily leave the filter over time (i.e., time to reach half the counts of the initial decay-corrected image ranged from <6 to >72 h). The majority of the image counts (approximately 70%) were seen in 2 distinctive areas in the filter. They corresponded to sites where the manufacturer used polyurethane adhesive to attach the fiberglass filter medium to the filter housing. CONCLUSION: (133)Xe contamination within the reusable internal bacteria filter of our ventilation system was easily detected by a survey meter and imaging. Although initial activities and surface radiation levels were low, radiation safety practices would dictate that a (133)Xe-contaminated bacteria filter be stored preferably in a fume hood until it cannot be distinguished from background before autoclaving or disposal.