Chronic Toxicity Studies Research Articles

Toxicity of Chlorantraniliprole and it's Formulated Product, Altacor®, to Individuals and Populations of Ceriodaphnia Dubia Richard.

The toxicity of the insecticide chlorantraniliprole and its formulated product Altacor® was determined for the Cladoceran, Ceriodaphnia dubia Richard. Acute toxicity (48h) and 21 d population studies were conducted. The hypothesis of this study was that these two compounds would have different toxicities. We conducted acute and chronic toxicity studies for each compound and compared the results to test this hypothesis. 48h LC50s (95% CL) for chlorantraniliprole and Altacor® were 8.5 (6.6-11.5) and 6.0 (3.7-9.0) µg chlorantraniliprole/L water, respectively. Therefore, chlorantraniliprole and Altacor® were equitoxic to C. dubia at LC50 based on overlap of the 95% CL. In the population study, chlorantraniliprole and Altacor® concentrations equivalent to the acute LC5, 10, 25, and 50 for each product were evaluated on populations of C. dubia. Number of individuals after 21 d was the endpoint evaluated. T-tests conducted at each LC value indicated that there was no significant difference in population size between these two products at each LC value evaluated. Previous studies show that toxicity can vary greatly between formulated and technical grade pesticides. However, our results show that chlorantraniliprole and its formulated product, Altacor® were equally toxic to C. dubia. Therefore, making assumptions about the toxicity of formulated and unformulated products is ill advised.

P21-11 Re-evaluating the need for chronic toxicity studies with therapeutic monoclonal antibodies, using a weight of evidence approach

Different government agencies operating in the European Union regulate different types of chemical products but all require testing for carcinogenicity to support applications for product marketing and commercialisation. A conference was held in Brussels in 2013 where representatives of the pharmaceutical, animal health, chemical and plant protection industries, together with representatives of regulatory agencies, universities and other stakeholders, met under the auspices of The European Partnership for Alternative Approaches to Animal Testing (EPAA) to discuss the varying requirements for carcinogenicity testing, and how these studies might be refined to improve hazard evaluation and risk assessment while implementing principles of the 3Rs (replacement, refinement and reduction in animal studies). While there are some similarities, the regulatory approaches in pharmaceutical, animal health, chemical and plant protection sectors have varying degrees of flexibility in requirements for carcinogenicity testing, to an extent reflecting concerns over the magnitude and duration of human exposure, either directly as in therapeutic exposure to pharmaceuticals, or indirectly through the ingestion of residues of veterinary drugs or plant protection chemicals. The article discusses these differences and other considerations for modified carcinogenicity testing paradigms on the basis of scientific and 3Rs approaches.

LP-35 Sub-chronic and Chronic toxicity study of Investigational Anti-Hypertensive Small Molecule KSD179019

LP-35 Sub-chronic and Chronic toxicity study of Investigational Anti-Hypertensive Small Molecule KSD179019

2D and 3D liver models

2D and 3D liver models

Acute and sub chronic toxicity studies with herbal pain relieving formula (Rhuleave-K™) in rats

Rhuleave-K™ is a proprietary combination of Curcuma longa extract, Boswellia serrata extract and black sesame seed oil. Acute toxicity was evaluated as per OECD guidelines 423. Rhuleave-K™ was fed at 2000 mg/kg to overnight fasted female rats. Clinical signs of abnormality and mortality was observed daily for 14 days. Sub-chronic toxicity was studied by feeding Rhuleave-K™ at 100, 500 and 1000 mg/kg/day to rats as per OECD guidelines 408. After 90 days feeding, hematological and biochemical parameters were analyzed. Histopathology of all the major organs was also studied. In the acute toxicity study, there was no clinical sign of toxicity in any of the rats at maximum dose of 2000 mg/kg. The LD50 was computed as >2000 mg/kg in rats. The repeated dosing of Rhuleave-K™ at the maximum dose level of 1000 mg/kg for 90 days did not induce any observable toxic effects in rats, when compared to its corresponding control. The hematology and biochemistry profiles of treated rats were similar to control animals and difference was non-significant (p > 0.05). The histopathology of major organs of all the control and treated animals was normal. In this study the NOAEL for Rhuleave-K™ was calculated as 1000 mg/kg daily in rats.

Evaluation of the Acute and Sub-Acute Oral Toxicity of Jaranol in Kunming Mice.

Background: Jaranol has shown a wide range of pharmacological activities; however, no study has yet examined in vivo toxicity. The study aimed to investigate the oral acute and sub-acute toxicity of jaranol in mice. Methods: The acute toxicity was determined by a single oral dose of jaranol (2000 mg/kg). Therein animal behaviour and mortality rate were observed for 14 days. The jaranol (50, 100 and 200 mg/kg BW·d−1) was given by gavage for 28 days daily in the sub-acute study. The mouse body weight (BW), organ weight, food, water intake, biochemical, haematological parameters, and histopathology were studied in acute and sub-acute toxicity. Results: During the acute toxicity test, a single oral dose (2000 mg/kg) jaranol did not cause significant alteration in majority of the hematological indices. However, jaranol decreased the level of serum alanine aminotransferase and aspartate aminotransferase. Those results showed that the oral lethal dose 50 (LD50) of jaranol was higher than 2000 mg/kg BW, regardless of sex. In repeated daily oral doses (50, 100 and 200 mg/kg BW·d−1), no mortality was recorded in the various experimental groups. The jaranol reduced body weight gain (200 mg/kg BW·d−1), the relative spleen weight (all doses) and serum alanine aminotransferase activity (200 mg/kg BW·d−1). On the other hand, jaranol significantly elevated red blood cell count (100 and 200 mg/kg BW·d−1) and serum creatinine levels (200 mg/kg BW·d−1). Histological study revealed that spleen bleeding was identified in 200 mg/kg jaranol-treated mice. Conclusion: Jaranol was relatively safe in Kunming Mice when repetitively administered orally in small doses for a prolonged period of time. We recommend more chronic toxicity studies and clinical trials on jaranol to ensure that its use is free of potential toxicity to humans.

Abstract 299: Self-assembling nanoparticles: A novel approach for targeted cancer treatment using tumor treating fields

Abstract Tumor Treating Fields (TTFields) are used in cancer treatment as they offer increased efficacy, safety, and low adverse effects. They have shown significant potential in pancreatic cancer (PC) clinical trials when combined with chemotherapy. It is pertinent to know that although TTFields-chemotherapy combination definitely enhances the treatment efficacy, it also results in chemotherapy induced severe adverse effects. To resolve this, we earlier reported development of Gemcitabine loaded self-assembling cationic-anionic polymer nanoparticles (S-CAP NPs) capable of achieving TTFields-triggered targeted drug release at the PC tumor site. Initially, four S-CAP NPs [C4 and C7 from chitosan- bovine serum albumin (Chitosan-BSA) S-CAP NPs; P5 and P8 from polyethylenimine- bovine serum albumin (PEI-BSA) S-CAP NPs] were shortlisted [Table 1 - particle size: ~ 200 nm; % encapsulation efficiency (EE): 55-65%]. For proof-of-concept studies, effect of TTFields on the % EE and % cumulative drug release of the S-CAP-NPs was investigated. The studies revealed (Table 1) significant reduction (p value < 0.05) in the % EE of S-CAP NPs within 48h TTFields treatment, confirming that the TTFields destabilize the NPs triggering targeted drug release. Similarly, In vitro drug release studies showed significant increase (p< 0.01) in rate of drug release from S-Cap NPs in presence of TTFields. The cell inhibition assay confirmed the synergism in cell apoptosis wherein PEI-BSA S-CAP NPs showed higher efficacy compared to chitosan-BSA S-CAP NPs. Further, additional stability studies continued over a period of 18 months confirmed stable formulations. In vivo acute and chronic toxicity studies are underway to identify the most optimum dose of S-CAP NPs for further in vivo efficacy studies. To summarize, S-CAP-NPs present a promising avenue to achieve targeted chemotherapy in conjunction with TTFields. This work is supported by AACR-Novocure tumor treating fields research fellowship. Table 1. Effect of TTFields on % encapsulation efficiency (n=3). Data represented as Mean ± SD. Batch Particle size (nm) Encapsulation efficiency (%) - 0h Encapsulation efficiency (%) - 24h - 150 KHZ Encapsulation efficiency (%) - 48h - 150 KHZ Chitosan-BSA S-CAP NPs C4 210.54 ± 38.96 61.26 ± 5.11 36.21 ± 3.53 24.86 ± 3.19 C7 215.67 ± 32.55 65.31 ± 5.84 32.19 ± 2.68 21.86 ± 4.64 PEI-BSA S-CAP NPs P5 198.29 ± 41.05 58.83 ± 3.33 27.66 ± 3.31 18.96 ± 3.96 P8 209.92 ± 31.33 64.31 ± 5.13 24.83 ± 4.29 19.42 ± 2.73 Citation Format: Preshita Prafulla Desai, Sunil Prabhu. Self-assembling nanoparticles: A novel approach for targeted cancer treatment using tumor treating fields [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 299.

Effect of Chronic Toxicity Studies of Sappan Wood Extract on The Kupffer Cells Number in Rats (Rattus novergicus)

Sappan wood (Caesalpinia sappan L.) is a plant used by the community to mix drinking. The antioxidant index of Sappan wood extract has a higher value than commercial antioxidants to counteract oxidative free radicals and improve the body's defense system. The use of Sappan wood drinking in society is often used in daily so it is necessary to do a chronic toxicity test to observe the response to prolonged use of the system in the body. Kupffer cells are an important part of the innate immune system, acting as “scavengers” and phagocytes. A study that aims to see the chronic toxicity studies of the ethanolic extract of Sappan wood on the number of Kuffer cells in the liver has been carried out on 60 Wistar rats divided into 12 groups. The group was the negative control group, doses 100 mg/kgBW, 200 mg/kgBW, 300 mg/kgBW, 400 mg/kgBW and 500 mg/kgBW of extract Sappan wood which were divided into male and female groups. Each group was given ethanol extract of Sappan wood for one year orally. The results showed a significant increase in Kupffer cells in the female group at a dose of 100 mg/kgBW and the male group at 200 mg/kgBW with significant values, respectively, p<0.001 p = 0.004.

Risk Assessment of the Pesticide Coragen 20 SC with the Active Substance Chlorantraniliprole

Coragen 20 SC is a new product in Norway containing the active substance chlorantraniliprole. It is applied for use in apples against codling moth (Cydia pomonella), apple fruit moth (Argyresthia conjugella) and free leaf living larvae. The Norwegian Scientific Committee for Food Safety (VKM) was asked by the Norwegian Food Safety Authority to perform a risk assessment on human health, environmental fate and ecotoxicological of the active substance and the product. The risk assessment of the product was approved at a meeting 11th of May 2010 by VKMs Scientific Panel on Pesticides (Panel 2). VKMs Panel 2 concludes as following: The product and the active substance have low acute oral, dermal and inhalation toxicities. Both are non-irritating to the skin, and no allergenic potential by skin contact were shown. Coragen was non-irritating to the eyes, while chlorantraniliprole showed a weak irritating potential. Chlorantraniliprole is not shown to have any genotoxic or carcinogenic potential, or to be teratogenic or toxic to the reproduction of female animals. The potential for testicular toxicity of chlorantraniliprole is unclear because the study design and the limited number of young dogs (2/sex/group) do not provide basis for a firm conclusion. No particular target organ for toxicity in any species in the sub-chronic and chronic toxicity studies was seen. The observed dose- and time dependent increased degree of microvesiculation in the zona fasiculata of the adrenal cortex in male rats, is however of uncertain biological significance. All test species (rat, mice, dog) showed physiological adaption to chlorantraniliprole administration (increased liver metabolism with induction of cytochrome P450 enzymes) which was manifested as increased liver weight and hepatocellular hypertrophy. In the chronic toxicity study in mice, the increased liver weight and hepatocellular hypertophy was accompanied with eosinofilic foci, which was assessed as an adverse effect. The no observed effect level (NOAEL) derived from this study serves as basis for calculations of values for acceptable daily intake (ADI) and acceptable operator exposure level (AOEL). 
 In Panel 2‘s opinion a sub-chronic study (90 days) with the technical material (E2Y45-282) including relevant concentration of the impurity IN-G2S78 should be performed. This would bring information on possible influence of the impurity on the toxicological profile of the technical material, and consequently on the assessment of the NOAELs in the various toxic studies. 
 The estimated risk for operator and for bystanders or for workers re-entering treated crops is assessed as minimal. 
 Chlorantraniliprole is persistent in soil with half live of about 1 year. The long half life indicates high potential for accumulation in soil after repeated use, which is confirmed by both model simulations and field studies. The Panel considers field data from the south of Europe not to be relevant for the Nordic conditions based on different climate conditions and soil properties contributing in different directions.
 The Panel considers that the existing documentation is not sufficient for a firm conclusion on the use of normalised field data for modelling purposes. However, the substance is persistent and expected to accumulate in soil. The Panel concludes that there is minimal risk for toxic effects on mammals, birds, bees, and microorganisms in soil due to chlorantraniliprole exposure with the proposed exposure regime. Panel concludes that there is “very high risk” for effects on in-field non target arthropods from chlorantraniliprole exposure. 
 For soil living invertebrates, earthworms seem rather insensitive to chlorantraniliprole and the Panel considers the toxic effects to be minimal. Panel considers the risk for toxic effects on soil living arthropods to be high. Crustaceans and insects larvae are the aquatic organisms most sensitive to chlorantraniliprole. The Panel concludes that there is a minimal risk of toxic effects on aquatic organisms due to exposure to chlorantraniliprole with the proposed application regime provided that a buffer zone of 30 m to surface water is applied.

Toxicity assessment of TiO2-conjugated Carbon-based nanohybrid material on a freshwater bioindicator cladoceran, Daphnia magna.

The application of nanocomposite materials fabricated from titanium dioxide nanoparticles (TiO2 NPs) and different carbon (C) allotropes have gained popularity in water treatment applications due to their synergistic properties. Studies to date have focused on simple forms of nanomaterials (NMs), however, with the technology development, there is a dramatic increase in production and application of these complex NMs which could result in toxicological impacts on organisms when released into aquatic environments. This raises serious concerns about their safety and the need to ascertain their potential adverse effects on aquatic organisms. While conjugated TiO2 NPs/carbon-based nanohybrids (TiO2/C-NHs) may exhibit enhanced photocatalytic activity, there is no research in the scientific community regarding their toxicological effects on D. magna, which are indicators of freshwater pollution. In this study, two under-represented TiO2/C-NHs (i.e., TiO2- conjugated carbon nanofiber (CNF), and TiO2-conjugated multi-walled carbon nanotube (CNT)) were investigated for their toxic effects on D. magna, through a series of acute toxicity tests with a set of sublethal biochemical biomarkers of oxidative stress. The lethal toxicity and oxidative stress formation of TiO2/C-NHs over 48h revealed a concentration-dependant increase in D. magna mortality. The primary mechanism identified was the generation of ROS, which was in line with toxicity results. Light microscopy and CytoViva® images visualized D. magna interaction with the NPs, which accumulated and appeared as dark materials in the lines of the gut tract. The collective results indicate that TiO2/C-NHs have the potential to cause an effect on freshwater organisms when released into the environment. However, the relevance of TiO2/C-NHs effects needs further chronic toxicity studies since they show promise to be used in nano-bioremediation materials to treat wastewaters.

Evaluation of Toxic Properties of New Glycopeptide Flavancin on Rats.

Glycopeptide antibiotics have side effects that limit their clinical use. In view of this, the development of glycopeptides with improved chemotherapeutic properties remains the main direction in the search for new antibacterial drugs. The objective of this study was to evaluate the toxicological characteristics of new semi-synthetic glycopeptide flavancin. Acute and chronic toxicity of antibiotic was evaluated in Wistar rats. The medium lethal dose (LD50) and the maximum tolerated doses (MTD) were calculated by the method of Litchfield and Wilcoxon. In the chronic toxicity study, the treatment regimen consisted of 15 daily intraperitoneal injections using two dosage levels: 6 and 10 mg/kg/day. Total doses were equivalent to MTD or LD50 of flavancin, respectively. The study included assessment of the body weight, hematological parameters, blood biochemical parameters, urinalysis, and pathomorphological evaluation of the internal organs. The results of the study demonstrated that no clinical-laboratory signs of toxicity were found after 15 daily injections of flavancin at a total dose close to the MTD or LD50. The pathomorphological study did not reveal any lesions on the organ structure of animals after low-dose administration of flavancin. Thus, flavancin favorably differs in terms of toxicological properties from the glycopeptides currently used in the clinic.

Chronic (52-week) oral toxicity study of herbal tea of Moringa stenopetala and Mentha spicata leaves formulation in Wistar albino rats

Background: Moringa stenopetala leaves have long been used to treat diabetes, hypertension, respiratory problems, and other diseases. The herbal formulation of Moringa stenopetala and Mentha spicata leaves was found to be more effective in lowering high blood pressure and blood sugar levels. Unlike its pharmacological properties, the long-term safety profile of this herbal formulation has not been investigated yet. Thus, this study investigated the long-term (chronic) oral toxicity of herbal tea of M. stenopetala and M. spicata leaves blended in rats. Methods: Wistar albino rats were randomly distributed into four groups (n = 10/sex/group), and then randomly assigned to a control group and three test groups. The control group (G I) received distilled water. The test groups (G II-IV) received 559.36, 1118.72, and 2237.44 mg/kg of herbal tea of M. stenopetala and M. spicata leaves blend respectively, for 360 consecutive days. During the treatment period, in-life parameters (mortality, clinical symptoms, body weight, and food intake) were evaluated. On the 361st day, hematological, serum biochemical, gross morphological, and histological parameters were investigated. Results: Throughout the 360-day treatment period, no herbal tea-related deaths, severe clinical symptoms, loss of body weight, or food intake were seen in any of the treated groups. Bodyweight, food consumption, organ weight, hematological, and serum biochemical findings showed no significant differences between the control and treated groups in both sexes. Macro-pathological and histopathological examinations of the major organs (liver, kidney, heart, pancreas, stomach, and spleen) revealed no herbal tea-related pathologic alterations. Conclusion: The findings indicate that long-term (360-days) oral administration of the herbal tea of M. stenopetala and M. spicata leaves blend is well tolerated by rats. Hence, it would be safe/low toxic up to a dose of 2237.44 mg/kg/day in chronic exposure.

The reproductive effects of the cancer chemotherapy agent, Carmofur, on Daphnia magna are mediated by its metabolite, 5-Fluorouracil.

Carmofur is an antineoplastic agent that inhibits ceramidase, a key enzyme in the sphingolipid pathway. Previous research suggests carmofur represses reproductive maturity in Daphnia magna. The purpose of this experiment was to confirm carmofur's effects on fecundity and reproductive maturity over two generations. A chronic toxicity test found reproductive maturity was delayed from 9 to 19 days by 0.80 μM carmofur with a 99.7% drop in reproduction, probably caused by delayed ovarian development. Second generation effects were even greater with 0% reproductive success at 0.40 μM. To our surprise, carmofur was not measured in the media by HPLC 24 h after exposure. Previous research indicated that carmofur is unstable in water and hydrolyzed into 5-fluorouracil (5-FU). Therefore, the chronic toxicity study was repeated with 5-FU and similar effects on reproductive maturity were observed at similar concentrations despite very different acute toxicities (48 h carmofur LC50 = 1.93 μM; 5-FU LC50 = 207 μM). 5-FU delayed reproductive maturity from 9 to 21 days with a 71.12% drop in reproduction at 0.80 μM and greater effects in the 2nd generation similar to carmofur. 5-FU was found stable in aquatic media and HPLC confirmed 5-FU was hydrolyzed from carmofur within 24 h. In conclusion, carmofur and 5-FU reduce fecundity because they delay reproductive maturity and ovarian development in Daphnia magna. We conclude that the reproductive effects observed after carmofur treatment are primarily mediated by its breakdown product, 5-FU. This further underscores the importance of measuring chemical concentrations and evaluating chemical metabolism and decomposition when determining toxicity, especially of chemotherapeutic agents.Clinical trials registration Not applicable.

English

Erythrina sacleuxii, one of five Erythrina species found in Kenya, is a stiffly multi-branched deciduous tree reported to have antiplasmodial, antifungal, anti-bacterial and anti-cancer activities. This study aimed at evaluating acute and sub-acute oral toxicity of E. sacleuxii roots ethanolic extract. Phytochemical analysis was conducted on Fourier transform infrared spectroscopy and Gas Chromatography-Mass Spectrometer. For acute and sub-acute toxicity studies 16 male rats n=4/group were used. The control group received 2% DMSO while treatment groups received 400, 1200 and 2400 mg/kg single dose (acute toxicity assay) and repeated for 14 days in subacute toxicity assay. Histopathological, serum biochemistry and hematological tests were analyzed. The results of FT-IR analysis recorded 12 dominant peaks, confirming availability of N-H, C-N, -C=C-, C-H, S=O, C-O and C-C functional groups. GC-MS analysis revealed presence of 17 bioactive phytochemicals mainly terpenoids and phenols. No toxic effect, organ toxicity or death was recorded in acute toxicity assay. In the subacute toxicity investigation, rats' behavior, gross pathology, and hematological parameters showed no significant changes. Biochemical analysis indicated an increase in alkaline phosphatase from 147 U/L at 400 mg/kg to 164 U/L at 2400 mg/kg however this was not statistically significant and no mortality was recorded. In comparison to the negative control, treatment groups’ gross and histopathology of livers and kidneys had no significant difference. These results demonstrate that 80% ethanolic extract of E. sacleuxii roots has no severe effects in both acute and subacute study. However, sub-chronic and chronic toxicity studies are necessary for further validation. Key words: Erythrina sacleuxii, acute and sub-acute studies, phytochemical analysis.

A retrospective analysis of honey bee (Apis mellifera) pesticide toxicity data.

Current USEPA ecological risk assessments for pesticide registration include a determination of potential risks to bees. Toxicity data are submitted to support these assessments and the USEPA maintains a large database containing acute and chronic toxicity data on adult and larval honey bees (Apis mellifera), which USEPA considers a surrogate for Apis and non-Apis bees. We compared these toxicity data to explore possible trends. This analysis indicated a significant correlation between acute contact and oral median lethal dose (LD50) values for adult honey bees (ρ = 0.74, p <0.0001). Using default EPA modeling assumptions, where exposure for an individual bee is roughly 12x lower through contact than through ingestion, the analysis indicates that the oral LD50 is similarly if not more protective of the contact LD50 for the majority of pesticides and modes of action evaluated. The analysis also provided evidence that compounds with a lower acute toxicity for adults through contact and oral exposure pathways may still be acutely toxic for larvae. The acute toxicity of herbicides and fungicides was higher for larvae relative to oral and contact toxicity for adult honey bees for the same compounds and the no observed adverse effect level (NOAEL) from chronic toxicity studies were lower for larvae relative to adults, indicating increased sensitivity of larvae. When comparing 8-day LD50 values between single dose larval acute studies to those derived from repeat dose 22-day larval chronic toxicity studies, the LD50 values derived from chronic studies were significantly lower than those from acute toxicity tests (Z = -37, p = 0.03).

Safety and efficacy of a feed additive consisting of sepiolite for all animal species (Sepiol S.A and Tolsa, S.A).

Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the re‐evaluation of the authorisation of sepiolite as a feed additive for all animal species. The FEEDAP Panel considered that sepiolite is unlikely to be absorbed. Harmful amounts of residues of any chemical component in edible tissues/products, as a consequence of the use of sepiolite as a feed additive, are not expected. Sepiolite is not genotoxic and does not induce any toxicity effects following oral administration and, therefore, it was considered safe for the consumers. The additive was considered safe for dairy cows at the recommended use level with a safety factor of 2.5. The conclusion was extrapolated to other dairy ruminants but owing to the lack of sufficient data, no conclusions can be drawn on the safety of the additive for the other target species/categories. Based on the results of a chronic inhalation toxicity study, the additive is considered a respiratory irritant. Owing to the dusting potential of the additive and its silica content, handling the additive was considered a risk by inhalation for the users. It is not irritant or corrosive to skin or eyes. Due to the nickel content, it is considered a skin and respiratory sensitiser. The additive was considered safe for the environment. The FEEDAP Panel concluded that sepiolite was efficacious as a thickener‐suspending agent, binder and anticaking agent in feed for all animal species under the proposed conditions of use.

Rethinking chronic toxicity and carcinogenicity assessment for agrochemicals project (ReCAAP): A reporting framework to support a weight of evidence safety assessment without long-term rodent bioassays

Rodent cancer bioassays have been long-required studies for regulatory assessment of human cancer hazard and risk. These studies use hundreds of animals, are resource intensive, and certain aspects of these studies have limited human relevance. The past 10 years have seen an exponential growth of new technologies with the potential to effectively evaluate human cancer hazard and risk while reducing, refining, or replacing animal use. To streamline and facilitate uptake of new technologies, a workgroup comprised of scientists from government, academia, non-governmental organizations, and industry stakeholders developed a framework for waiver rationales of rodent cancer bioassays for consideration in agrochemical safety assessment. The workgroup used an iterative approach, incorporating regulatory agency feedback, and identifying critical information to be considered in a risk assessment-based weight of evidence determination of the need for rodent cancer bioassays. The reporting framework described herein was developed to support a chronic toxicity and carcinogenicity study waiver rationale, which includes information on use pattern(s), exposure scenario(s), pesticidal mode-of-action, physicochemical properties, metabolism, toxicokinetics, toxicological data including mechanistic data, and chemical read-across from similar registered pesticides. The framework could also be applied to endpoints other than chronic toxicity and carcinogenicity, and for chemicals other than agrochemicals.

Chitosan-based lifefoam improves survival in lethal noncompressible abdominal bleeding in swine

BackgroundIn military combat settings, noncompressible closed cavity exsanguination is the leading cause of potentially survivable deaths, with no effective treatment available at point of injury. The aim of this study was to assess whether an expanding foam based on hydrophobically modified chitosan (hm-chitosan) may be used as a locally injectable hemostatic agent for the treatment of noncompressible bleeding in a swine model. MethodsA closed-cavity, grade V hepato-portal injury was created in all animals resulting in massive noncoagulopathic, noncompressible bleeding. Animals received either fluid resuscitation alone (control, n = 8) or fluid resuscitation plus intraperitoneal hm-chitosan agent through an umbilical port (experimental, n = 18). The experiment was terminated at 180 minutes or death (defined as end-tidal CO2 <8mmHg or mean arterial pressure [MAP] <15mmHg), whichever came first. ResultsAll animals had profound hypotension and experienced a near-arrest from hypovolemic shock (mean MAP = 24 mmHg at 10 minutes). Mean survival time was higher than 150 minutes in the experimental arm versus 27 minutes in the control arm (P < .001). Three-hour survival was 72% in the experimental group and 0% in the control group (P = .002). Hm-chitosan stabilized rising lactate, preventing acute lethal acidosis. MAP improved drastically after deployment of the hm-chitosan and was preserved at 60 mmHg throughout the 3 hours. Postmortem examination was performed in all animals and the hepatoportal injuries were anatomically similar. ConclusionIntraperitoneal administration of hm-chitosan-based foam for massive, noncompressible abdominal bleeding improves survival in a lethal, closed-cavity swine model. Chronic safety and toxicity studies are required.

Evaluation of Repeated Dose (Sub Chronic) Toxicity Study of Shilasindur Prepared by Two Different Methods Along with Its Characterization and Antimicrobial Activity

Introduction: Rasaushadhies (metal and mineral) preparations are unique preparations that include Bhasma, herbo-mineral preparations and Kupipakwa rasayana hold a significant place in Ayurvedic pharmaceutics. Nearly 70 % of formulations include a combination of one or more metallic/mineral with several herbs which have a supporting role in improving efficacy, relieving symptoms of the disease and to achieve long and healthy life. Shilasindur is a one of the Kupipakwa rasayana containing mercury, sulphur and arsenic disulphide and indicated in skin disorders, respiratory tract disorders and other infectious diseases.&#x0D; Aim and Objectives: Evaluation of repeated dose (Sub chronic) toxicity study of Shilasindur along with its characterization and antimicrobial activity.&#x0D; Materials &amp; Methods: Shilasindur will be prepared in three batches as per classical reference and by using Electric Muffle furnace. The prepared formulation will be assessed with Organoleptic characters, physicochemical parameters and ICP – AES (Inductively coupled plasma atomic emission spectroscopy), FTIR (Fourier-transform infrared spectroscopy), XRD (X-Ray Diffraction). Sub chronic toxicity study will be carried out according to the OECD guidelines 408 and 452 as well as CCRAS guidelines of safety and toxicity. Epsilometer test (E-test) will be used to check antimicrobial activity of Shilasindur.&#x0D; Results: The analytical parameters will be assessed to establish the pharmaceutical standardization. Toxicity study will be done in three doses to ensure its safety. Statistical analysis will be done by applying unpaired t-test and ANOVA test.&#x0D; Conclusion: If significant positive results obtained in this work then it will be a valuable contribution and evidence that the drug Shilasindur is safe for consumption at treatment doses.

Sensitivity of Buff-Tailed Bumblebee (Bombus terrestris L.) to Insecticides with Different Mode of Action.

Simple SummarySeveral neonicotinoid insecticides that were once widely used for pest control are currently banned for outdoor use in the European Union (EU) because they pose a risk to bees. This restriction meant that farmers had to look for alternatives for pest management and use known insecticides or new substances with supposedly more bee-friendly characteristics. We evaluated the toxicity of six insecticides on buff-tailed bumblebee workers (Bombus terrestris): two banned neonicotinoids (imidacloprid, thiacloprid), two pyrethroids (deltamethrin, esfenvalerate), one sulfoximine (sulfoxaflor) and a microbial insecticide based on Bacillus thuringiensis toxins, which are present in genetically modified (Bt) maize. The results obtained show that certain insecticides in use have higher acute toxicity to B. terrestris than some of the banned neonicotinoids.Systemic insecticides are recognized as one of the drivers of the worldwide bee decline as they are exposed to them through multiple pathways. Specifically, neonicotinoids, some of which are banned for outdoor use in the European Union (EU), have been pointed out as a major cause of bee collapse. Thus, farmers have had to look for alternatives for pest control and use known insecticides or new substances reportedly less harmful to bees. We evaluated the oral acute toxicity of six insecticides (three of them systemic: imidacloprid, thiacloprid and sulfoxaflor) with four different modes of action on buff-tailed bumblebee workers (Bombus terrestris): two banned neonicotinoids (imidacloprid, thiacloprid), two pyrethroids (deltamethrin, esfenvalerate), one sulfoximine (sulfoxaflor) and a microbial insecticide based on Bacillus thuringiensis toxins, present in genetically modified (Bt) maize. The microbial insecticide only caused mortality to bumblebee workers at extremely high concentrations, so it is expected that Bt maize does not pose a risk to them. The toxicity of the other five insecticides on bumblebees was, from highest to lowest: imidacloprid, sulfoxaflor, deltamethrin, esfenvalerate and thiacloprid. This outcome suggests that certain insecticides in use are more toxic to B. terrestris than some banned neonicotinoids. Further chronic toxicity studies, under realistic conditions, are necessary for a proper risk assessment.