Specific Target Organ Toxicity Research Articles

Paired Liver:Plasma PFAS Concentration Ratios from Adolescents in the Teen-LABS Study and Derivation of Empirical and Mass Balance Models to Predict and Explain Liver PFAS Accumulation.

Animal studies have pointed at the liver as a hotspot for per- and polyfluoroalkyl substances (PFAS) accumulation and toxicity; however, these findings have not been replicated in human populations. We measured concentrations of seven PFAS in matched liver and plasma samples collected at the time of bariatric surgery from 64 adolescents in the Teen-Longitudinal Assessment of Bariatric Surgery (Teen-LABS) study. Liver:plasma concentration ratios were perfectly explained (r2 > 0.99) in a multilinear regression (MLR) model based on toxicokinetic (TK) descriptors consisting of binding to tissue constituents and membrane permeabilities. Of the seven matched plasma and liver PFAS concentrations compared in this study, the liver:plasma concentration ratio of perfluoroheptanoic acid (PFHpA) was considerably higher than the liver:plasma concentration ratio of other PFAS congeners. Comparing the MLR model with an equilibrium mass balance model (MBM) suggested that complex kinetic transport processes are driving the unexpectedly high liver:plasma concentration ratio of PFHpA. Intratissue MBM modeling pointed to membrane lipids as the tissue constituents that drive the liver accumulation of long-chain, hydrophobic PFAS, whereas albumin binding of hydrophobic PFAS dominated PFAS distribution in plasma. The liver:plasma concentration data set, empirical MLR model, and mechanistic MBM modeling allow the prediction of liver from plasma concentrations measured in human cohort studies. Our study demonstrates that combining biomonitoring data with mechanistic modeling can identify underlying mechanisms of internal distribution and specific target organ toxicity of PFAS in humans.

Pesticide drift mitigation measures appear to reduce contamination of non-agricultural areas, but hazards to humans and the environment remain

Pesticide drift onto non-agricultural land is a common problem in intensively farmed regions, and national action plans have been established across Europe to prevent it. Here, we analyzed official data on pesticide residues in grass samples collected over six years to determine whether implemented measures to reduce pesticide drift were effective. We used 306 samples collected between 2014 and 2020 on non-agricultural land in one of the most intensively managed apple and wine growing regions in Europe, the Autonomous Province of Bolzano-South Tyrol, Italy. Samples were analyzed for up to 314 substances by gas chromatography and mass spectrometry. Percentage of sites with multiple pesticides and number of pesticides decreased between 2014 and 2020. Fungicides were most often detected, with fluazinam found on 74 % and captan on 60 % of the contaminated sites (53 sites out of a total of 88 sites were contaminated). The most frequently found insecticide, phosmet, was detected in 49 % of the contaminated sites. Only one herbicide, oxadiazon, was detected in <1 % of the sites; glyphosate was not analyzed. The percentage of residues with human hazard properties increased significantly across years regarding reproductive toxicity (from 21 % of the detected substances in 2014 to 88 % in 2020) and specific target organ toxicity (0 % in 2014 to 21 % in 2020). Percentages of substances associated with endocrine-disruption (89 % of substances across years) or carcinogenic properties (45 % of substances across years) remained constant. The percentage of sites where concentrations in grass samples exceeded the surrogate maximum residue levels (MRLs) for lettuce also remained constant. Potential ecotoxicological hazards of detected residues regarding acute contact toxicity to honeybees remained high over the study years, while the acute and chronic toxicity to earthworms decreased. Our results suggest that while drift mitigation measures contributed some reduction in pesticide contamination, they were not sufficient to eliminate substantial risks to human health and the environment in nontarget areas.

Inflammation as a Key Outcome Pathway in Particle Induced Effects in the Lung.

Inflammation is considered a key event in the pathology of many chronic diseases, including pulmonary and systemic particle induced effects. In addition, inflammation is now considered as the key response in standard setting for poorly-soluble low toxicity (PSLT) particles and also the critical endpoint to screen for in OECD based sub-chronic animal inhalation testing protocols. During Particles & Health 2021, an afternoon session was dedicated to the subject and a brief summary of the most important messages are summarized in this paper. In the first part of this session, two speakers (Prof. Lison and Dr Duffin) provided state of the art insight into different aspects and sequels to (persistent) inflammation as a protective or adverse response. Most recent insights on the role of different macrophage cell types were presented as well as perspectives and data provided by inflammatory pathways in humans, such as in asthma and COPD. A brief review of the expert workshop on PSLT particles focusing on the regulatory impact of using persistent inflammation as a key outcome was provided by Kevin Driscoll. The second part of the session focused on the outcomes that are associated with inflammation in animal studies, with an emphasis by Drs. Harkema (Michigan State) and Weber (Anapath) on cell proliferation and other pathologies that need to be considered when comparing human and animal responses, such as outcomes from 14- or 28 day inhalation studies used for specific target organ toxicity classification.

Reducing overall herbicide use may reduce risks to humans but increase toxic loads to honeybees, earthworms and birds

BackgroundPesticide use has been associated with risks for human health and an overall decline in biodiversity. Although herbicides are the most commonly used pesticides worldwide, they have received less attention in this debate. We investigated the extent to which long-term trends in herbicide use in Austria influence potential toxic exposures to non-target organisms and potential risks to humans. We analyzed official sales data of 101 herbicide active ingredients (AIs) approved in Austria between 2010 and 2019 regarding their ecotoxicological properties based on lethal doses (LD50 and LC50) weighed by their persistence in the environment (DT50) for honeybees (Apis mellifera), earthworms (Eisenia fetida), and birds (Serinus serinus). Human health risks were qualitatively assessed based on official hazard statements for the AIs used.ResultsIn Austria, herbicide amounts sold decreased significantly by 24% from 1480 to 1123 tonnes between 2010 and 2019. This also led to a considerable decrease in the amounts of AIs classified by H-statements of the EU Pesticides Database: − 71% acute inhalation toxicity, − 58% reproductive toxicity,− 47% specific target organ toxicity. Yet, 36% of herbicides used were still classified as highly hazardous pesticides according to the Pesticide Action Network. Surprisingly, over the same period, toxic loads to honeybees increased by 487% (oral exposure), while lethal toxic loads to earthworms increased by 498%, and to birds by 580%. This can be attributed to a shift toward the use of more acutely toxic and especially more persistent AIs. The most problematic AI for honeybees, earthworms, birds and humans was the highly persistent diquat. The further ranking of the most toxic herbicides varied considerably depending on the organism. It is important to note that this toxic load assessment, like official environmental risk assessments, evaluates the potential risk but not the actual fatalities or real-world exposure.ConclusionsOur results show a trade-off between herbicide amounts and toxicological hazards to humans and other non-target organisms. These interdependencies need to be considered when implementing pesticide reduction targets to protect public health and biodiversity, such as the EU´s “farm-to-fork” strategy, which aims to reduce the amounts and risks of synthetic pesticides.

The impact on classifications for carcinogenicity, mutagenicity, reproductive and specific target organ toxicity after repeated exposure in the first ten years of the REACH regulation

The present study primarily aims at informing regulators and policy makers in Europe and examines the evolution of self-classifications and study availability for the endpoints of carcinogenicity, mutagenicity, reproductive toxicity (CMR) and specific target organ toxicity after repeated exposure (STOT RE) for the first ten years of the REACH legislation. Our knowledge on chemical safety keeps increasing due to the registration obligations under REACH, in combination with proactive actions by registrants and regulatory actions by Authorities, which jointly lead to new testing and critical reassessment of existing studies. The improvements become evident by the constant increase in the number of substances that are self-classified by the registrants for human health endpoints. Moreover, there is a slow but steady increase in the number of substances for which there is at least one experimental study available for the human health endpoints in scope of this analysis. However, the increase is slow given the generally limited data availability at the beginning of REACH. Manual examination of about 350 classified substances reveals that the impact of newly generated data and regulatory action by Authorities is greater for reproductive toxicity than for carcinogenicity or mutagenicity, reflecting the strengthening of the information requirements for reproductive toxicity with the introduction of REACH. The results of the study should inform regulators and policy makers at EU and national level in the discussion on potential changes to information requirements or testing strategies under REACH.

VOCs in cleaning products used in age care and social facilities: Identification of hazardous substances

Prior to the nationwide survey on indoor air quality in 100 age care and social facilities across France, which is planned for 2019, a questionnaire was sent by email to the directors of all 18,432 institutions of these types to determine the potential emission sources of indoor pollutants. Among these, cleaning practices and cleaning products were targeted. The questionnaire included items regarding cleaning frequency and periods, cleaning techniques, storage areas, the use of air fresheners and the commercial names of the products used. A total of 2140 questionnaires were collected and processed, and 1109 cleaning products, along with their commercial names, were listed. From them, 341 different products were identified, for which 299 safety data sheets (SDSs) were available and analyzed. A total of 216 different chemical substances were identified in the SDSs. The boiling points were retrieved to classify the substances according to their volatility. Finally, information regarding their hazard classification was collected, and six categories of human health effects were considered. A total of 41 chemical substances were classified, among which 10 are carcinogenic, 6 are mutagenic, 1 has reproductive toxicity, 3 have specific target organ toxicity - repeat exposure, 19 are possible endocrine disruptors, 14 are skin sensitizers, and 2 are respiratory sensitizers. Of these 41 chemicals, 51% (n = 21) are volatile organic compounds (VOCs). Despite the limitations of using SDSs, which do not report the exact compositions of the products, this study shows that a large spectrum of volatile substances may be emitted from cleaning products used in age care and social facilities, which may have a potential impact on the indoor air quality.

아급성흡입독성시험을 이용한 3-Methylpentane의 GHS 분류·표시

본 연구는 3-methylpentane에 대한 흡입유해성을 평가하여 국제연합에서 정하는 화학물질의 분류 및 표지에 관한 세계조화시스템(Globally harmonized system, GHS)지침 및 고용노동부고시 제2013-37호에 따른 3-methylpentane의 화학물질 분류 표시 자료를 생산하기 위하여 OECD 화학물질 시험가이드라인 아급성흡입독성시험 TG 412(Subacute inhalation toxicity) 시험법에 따라 수행하였다. 본 연구를 위하여 6주령의 랫드(Rat)를 도입하여 1주간 순화시킨 후 암수 각각 대조군 5마리, 저농도군(284 ppm) 5마리, 중농도군(1,135 ppm) 5마리, 고농도군(4,540 ppm) 5마리 등으로 군을 구성하여 일일 6시간, 주 5일, 4주 동안 시험물질을 랫드에 전신으로 노출시켰다. 시험물질 노출을 종료하고 2주 후 시험동물을 희생하여 시험물질에 의한 시험동물의 영향을 평가하였다. 사료섭취량 변화, 체중 변화, 임상관찰, 혈액검사, 부검 소견, 장기무게 측정, 조직병리검사 등 모든 시험결과에서 시험물질에 의한 영향은 나타나지 않아 3-methylpentane의 무유해영향농도는 암수 모두 4,540 ppm이상으로 판단되어 세계조화시스템(GHS) 지침 및 고용노동부고시 제2013-37호(화학물질의 분류 표시 및 물질안전보건에 관한 기준)의 특정표적장기독성(반복노출) 구분 표시 물질에 해당하지 않은 물질로 판단되었다. Objectives : The purpose of this study was to obtain information regarding Globally Harmonized System(GHS) classification and health hazards that may result from a 4 weeks inhalation exposure of 3-Methylpentane in Sprague-Dawley rats. Methods : The testing method was conducted in accordance with OECD guidelines for the testing of chemicals No. 412(Subacute Inhalation Toxicity). The Rats were divided into 4 groups(5 male and 5 female rats in each group) and exposed to 0 ppm, 284 ppm, 1,135 ppm, 4,540 ppm 3-Methylpentane in each exposure chamber for 6 h/day, 5 days/week, for 4 weeks. After two weeks, the test animals were autopsied and carried out blood test and biochemical tests and histopathological examination. We used PRISTIMA (Toxicology data management system) to confirm the system and to have confidence of the raw data. Results : No death and particular clinical presentation including weight change and change of feed rate was observed. Relationship between dose, gender and response was also not significantly changed in hematologic examination, biochemical examination of blood and blood coagulation time. The histopathologic lesions caused by the test substance did not appear. Conclusions : NOAEL(No Observable Adverse Effect Level) of 3-Methylpentane is more than 4,540 ppm in male group and female group and the Ministry of Employment and Labor Guidance Announcement No. 2013-37(criteria for the classification marks and Safety of Chemicals) Specific target organ toxicity(repeated exposure) was determined with a substance that is not the separator material.

SWeRF—A Method for Estimating the Relevant Fine Particle Fraction in Bulk Materials for Classification and Labelling Purposes

In accordance with the European regulation for classification, labelling and packaging of substances and mixtures (CLP) as well as the criteria as set out in the Globally Harmonized System (GHS), fine fraction of crystalline silica (CS) has been classified as a specific target organ toxicity, the specific organ in this case being the lung. Generic cut-off values for products containing a fine fraction of CS trigger the need for a method for the quantification of the fine fraction of CS in bulk materials. This article describes the so-called SWeRF method, the size-weighted relevant fine fraction. The SWeRF method combines the particle size distribution of a powder with probability factors from the EN 481 standard and allows the relevant fine fraction of a material to be calculated. The SWeRF method has been validated with a number of industrial minerals. This will enable manufacturers and blenders to apply the CLP and GHS criteria for the classification of mineral products containing RCS a fine fraction of CS.

A Nonclinical Safety Assessment of MnTE-2-PyP, a Manganese Porphyrin

Manganese (III) meso-tetrakis(N-ethylpyridinium-2-yl)porphyrin (MnTE-2-PyP or BMX-010; CASRN 219818-60-7) is a manganese porphyrin compound developed as a potential drug substance for use as a radioprotective and for the ex vivo treatment of cells, tissues, and organs intended for transplantation. In preparation for an investigational new drug filing, a full good laboratory practice nonclinical safety assessment was conducted in order to evaluate the safety of MnTE-2-PyP and included the performance of in vitro genotoxicity studies, local tissue tolerance evaluation, safety pharmacology core battery studies, and single- and repeat-dose intravenous (iv) toxicity studies in mice and monkeys. The MnTE-2-PyP was determined not to be genotoxic or hemolytic, did not demonstrate flocculation or elicit adverse pharmacologic effects on respiration, the central nervous system (CNS), and had limited transitory effects on the cardiovascular system only at levels well above the therapeutic target dose. The intended iv clinical solution did not cause venous irritation in rabbits. The no observed adverse effect level (NOAEL) in mice was determined to be 10 mg/kg/day after 18 consecutive days of bolus iv dosing once daily in the morning. The NOAEL in monkeys after 14 days of bolus iv dosing in the morning was determined to be 5 mg/kg/day. At doses relevant to clinical use in humans, neither study revealed any indication of any specific target organ toxicity, including the classic heme porphyrin kidney, liver, CNS, or cardiac toxicities, or manganese toxicity. Mortality seen shortly after dosing in individual animals at higher doses was not accompanied by any organ or clinical pathology indications, suggesting a functional pharmacological-mediated effect. Based on the results of these studies, a conservative safe initial starting clinical dose of 5.0 mg (0.083 mg/kg in a 60 kg adult) was proposed for the initiation of human trials. Because of patent life issues, use of MnTE-2-PyP as a transplantation aid or radioprotective agent is not currently being pursued past the preclinical stages. It serves as a model for the clinical development of this class of drugs.

Subchronic Inhalation Toxicity Study of n-pentane in Rats

ObjectivesThis study was conducted in order to obtain information concerning the health hazards that may result from a 13 week inhalation exposure of n-pentane in Sprague-Dawley rats.MethodsThis study was conducted in accordance with the Organization for Economic Co-operation and Development (OECD) guidelines for the testing of chemicals No. 413 'Subchronic inhalation toxicity: 90-day study (as revised in 2009)'. The rats were divided into 4 groups (10 male and 10 female rats in each group), and were exposed to 0, 340, 1,530, and 6,885 ppm n-pentane in each exposure chamber for 6 hour/day, 5 days/week, for 13 weeks. All of the rats were sacrificed at the end of the treatment period. During the test period, clinical signs, mortality, body weights, food consumption, ophthalmoscopy, locomotion activity, urinalysis, hematology, serum biochemistry, gross findings, organ weights, and histopathology were assessed.ResultsDuring the period of testing, there were no treatment related effects on the clinical findings, body weight, food consumption, ophthalmoscopy, urinalysis, hematology, serum biochemistry, gross findings, relative organ weight, and histopathological findings.ConclusionThe no-observable-adverse-effect level (NOAEL) of n-pentane is evaluated as being more than 6,885 ppm (20.3 mg/L) in both male and female rats. n-pentane was not a classified specific target organ toxicity in the globally harmonized classification system (GHS).

Frontiers in Preclinical Safety Biomarkers

The measurement of plasma microRNAs (miRNAs) and messenger RNAs (mRNAs) is the most recent effort to identify novel biomarkers in preclinical safety. These genomic markers often display tissue-specific expression, may be released from the tissues into the plasma during toxic events, change early and with high magnitude in tissues and in the blood during specific organ toxicities, and can be measured using multiplex formats. Their validation as biomarkers has been challenged by the technical difficulties. In particular, the concentration of miRNAs in the plasma depends on contamination by miRNAs originating from blood cells and platelets, and the relative fraction of miRNAs in complexes with Argonaute 2, high-density lipoproteins, and in exosomes and microvesicles. In spite of these hurdles, considerable progress has recently been made in assessing the potential value of miRNAs in the clinic, especially in cancer patients and cardiovascular diseases. The future of miRNAs and mRNAs as biomarkers of disease and organ toxicity depends on our ability to characterize their kinetics and to establish robust collection and measurement methods. This review covers the basic biology of miRNAs and the published literature on the use of miRNAs and mRNAs as biomarkers of specific target organ toxicity.

Evaluation of information in nanomaterial safety data sheets and development of international standard for guidance on preparation of nanomaterial safety data sheets

Safety data sheets (SDSs) and labelling are the basic hazard communication tools for hazardous chemicals as regards their manufacture, storage, transport and other handling activities. Thus, in the context of the growing use of nanomaterials and nanomaterial-containing materials, this study evaluated the information provided in 97 nanomaterial-related SDSs according to the criteria set by the GHS (Globally Harmonized System of Classification and Labelling of Chemicals) and found that most of the SDSs did not include sufficient information on the safety of nanomaterials, such as their toxicity and physicochemical properties. The reasons for this lack of information in the nanomaterial SDSs can mainly be attributed to (1) a lack of toxicity and physicochemical property information on nanomaterials, (2) unawareness of the effectiveness of conventional exposure controls, such as local exhaust ventilation and encapsulation, and personal protective equipment (PPE), in protecting against nanomaterial exposure, (3) a lack of information on emergency and firefighting measures and (4) a lack of knowledge on how existing regulations apply to nanomaterials. Therefore, to create a consistent standard for the information provided on safety, health and environmental matters for manufactured nanomaterial-containing products, guidance for the preparation of nanomaterial-specific SDSs, including both nanomaterials and mixtures of nanomaterials with conventional non-nanoscale materials, was recently initiated by the ISO TC 229. Their guidance, in the form of a technical report, recommends that nanomaterial-related SDSs should be prepared based on a precautionary approach in terms of the toxicity and other risks associated with the nanomaterial contents within the mixture in question. One of the key recommendations in the technical report is to include additional physicochemical properties, including the particle size (average and range), size distribution aggregation/agglomeration state, shape and aspect ratio, crystallinity, specific surface area, dispersibility and dustiness, which help to distinguish the characteristics of nanomaterials from those of non-nanoscale materials. The technical report also recommends the preparation of SDSs for all nanomaterials and mixtures that meet the GHS criteria for physical, health or environmental hazards, and for all mixtures containing nanomaterials that meet the criteria for carcinogenic, toxic to reproduction or specific target organ toxicity in concentrations exceeding the cut-off limits for an SDS specified by the criteria for mixtures. Finally, the technical report recommends that SDSs be prepared for all nanomaterials, unless there is evidence that they are not hazardous.

Acute and Subchronic Inhalation Toxicity of n-Octane in Rats

ObjectivesWe have investigated the toxic effects of the inhalation of subchronic and acute levels of n-octane.MethodsThe rats were exposed to n-octane of 0, 2.34, 11.68 and 23.36 mg/L (n = 5 rats/group/gender) in an acute inhalation test (Organization for Economic Co-operation and Development (OECD) TG 403), or to 0, 0.93, 2.62 and 7.48 mg/L (n = 10 rats/group/gender) for a subchronic inhalation test (OECE TG 413), to establish a national chemical management system consistent with the Globally Harmonized Classification System (GHS).ResultsAcutely-exposed rats became lethargic but recovered following discontinuation of inhalation. Other clinical symptoms such as change of body weight and autopsy finds were absent. The LC50 for the acute inhalation toxicity of n-octane was determined to exceed 23.36 mg/L and the GHS category was 'not grouping'. Subchronically-treated rats displayed no significant clinical and histopathological differences from untreated controls; also, target organs were affected hematologically, biochemically and pathologically. Therefore, the no observable adverse effect level was indicated as exceeding 7.48 mg/L and the GHS category was 'not grouping' for the specific target organ toxicity upon repeated exposure.ConclusionHowever, n-octane exposure should be controlled to be below the American Conference of Industrial Hygienists recommendation (300 ppm) to prevent inhalation-related adverse health effects of workers.

The preclinical toxicology of salmeterol hydroxynaphthoate

An extensive toxicology programme on salmeterol hydroxynaphthoate (Serevent), a marketed long-acting beta(2)-adrenoceptor agonist, has been carried out. The studies evaluated both the local (respiratory tract) and systemic tolerance to single and repeated dosing, effects on all stages of reproduction, as well as the genotoxic and oncogenic potential. High acute doses were well tolerated and caused no specific target organ toxicity. In repeat dose studies, animals tolerated salmeterol very well both locally and systemically. No significant effects on the respiratory tract of dogs were seen and only minor laryngeal changes, typical of those occurring with many inhaled medicines, were noted in rats. The high systemic concentrations achieved resulted in a number of changes that are considered to be the result of excessive and prolonged beta( 2)-adrenoceptor stimulation. These included tachycardia, skeletal muscle hypertrophy and minor haematological and blood biochemical changes in general toxicity studies, foetal effects in rabbit organogenesis studies and increased incidences of smooth muscle tumours of the mesovarium in the rat and of the uterus in the mouse oncogenicity studies. Salmeterol showed no evidence of any genotoxic potential. Results of the extensive toxicology programme provide good assurance of the safety for the inhaled use of salmeterol in patients; this has ben confirmed by many years of clinical experience during its development and marketing.

化学物質の危険有害性に関する分類・表示の世界調和システム（GHS）とわが国における実施について

The Globally Harmonized System of Classification and Labelling of Chemicals (GHS) is a set of recommendations by the United Nations, first issued in 2003 as a communication tool for the sound management of chemicals, comprising harmonized classification criteria for physical, health and environmental hazards, a unified format for material safety data sheets (MSDS), and labeling elements including pictograms and hazard statements preassigned to each classification category. The GHS has been introduced into Japan and implemented in the regulatory framework for chemical safety. The Japanese Industrial Standards (JIS) adopted the GHS, and the GHS-based JIS rules have become the Japanese standards for labels and MSDS. The use of the JIS format for labels and MSDS is recommended by several competent authorities in Japan although mostly on a voluntary basis. In the workplace, however, GHS-based JIS labels and MSDS have become legal requirements by the Industrial Safety and Health Law since 2006; namely, issuing MSDS in such a format is mandatory for the 640 specified chemicals and also labeling for the 99 targeted chemicals*. Although the GHS provides definitions and classification criteria for 10 classes of health hazards (acute toxicity, skin and eye corrosion/irritation, sensitization, germ cell mutagenicity, carcinogenicity, reproductive toxicity, specific target organ toxicity single/repeated exposures, and aspiration hazard), it does not provide actual classification of chemicals, so that competent authorities and industries need to classify a number of chemicals and/or mixtures. Weight-of-evidence judgment and/or expert judgment would be necessary in many cases. In this paper, the outline of the GHS classification is described and problems of the GHS and its implementation are discussed.

Pharmacokinetic and toxicology assessment of INTERCEPT (S-59 and UVA treated) platelets.

The pathogen inactivation process developed by Cerus and Baxter Healthcare Corporations uses the psoralen, S-59 (amotosalen) in an ex vivo photochemical treatment (PCT) process to inactivate viruses, bacteria, protozoans, and leukocytes in platelet concentrates and plasma. Studies were performed by intravenous infusion of S-59 PCT formulations +/- compound adsorption device (CAD) treatment and with non-UVA illuminated S-59, using doses that were multiples of potential clinical exposures. The studies comprised full pharmacokinetic, single- and repeated-dose (up to 13 weeks duration) toxicity, safety pharmacology (CNS, renal, and cardiovascular), reproductive toxicity, genotoxicity, carcinogenicity testing in the p53(+/-) mouse, vein irritation, and phototoxicity. No specific target organ toxicity (clinical or histopathological), reproductive toxicity, or carcinogenicity was observed. S-59 and/or PCT formulations demonstrated CNS, ECG, and phototoxicity only at supraclinical doses. Based on the extremely large safety margins (>30,000-fold expected clinical exposures), the CNS and ECG observations are not considered to have any toxicological relevance. Additionally, after a complete assessment, mutagenicity and phototoxicity results are not considered relevant for the proposed use of INTERCEPT platelets. Thus, the results of an extensive series of in vitro and in vivo studies have not demonstrated any toxicologically relevant effects of platelet concentrates prepared by the INTERCEPT system.

Preclinical safety of a nucleic acid-targeted helinx TM compound: A clinical perspective

Helinx ™ technology (Cerus Corp, Concord, CA) uses amotosalen HCl (S-59) and ultraviolet A (UVA) light in an ex vivo photochemical treatment (PCT) to inactivate viruses, bacteria, and leukocytes in platelet concentrates while preserving therapeutic function. A comprehensive preclinical safety program was conducted, which included carcinogenicity, single-dose and multiple-dose (up to 13 weeks' duration) toxicity, safety pharmacology (central nervous system [CNS], renal, and cardiovascular), reproductive toxicity, genotoxicity, vein irritation, phototoxicity, and toxicokinetic testing. The results of the toxicokinetic analyses indicated that the test articles provided large multiples of the clinical exposure to S-59, whether the comparison was based on dose, maximum plasma concentration, or area under the concentration-time curve. No specific target organ toxicity, reproductive toxicity, or carcinogenicity was observed. S-59 and/or PCT formulations demonstrated CNS toxicity, electrocardiographic (ECG) effects, and phototoxicity at supraclinical doses. On the basis of the extremely large safety margins, the CNS and ECG observations (at > 30,000-fold the expected clinical exposure) as well as the results of genotoxicity and phototoxicity studies are not considered to be of toxicological relevance. The results of an extensive series of studies have thus demonstrated no toxicologically relevant effects of platelets treated with Helinx technology.

Evaluation of the chronic toxicity and oncogenicity of N,N-diethyl-m-toluamide (DEET).

Chronic toxicity and/or oncogenicity studies were conducted in rats, mice, and dogs with the insect repellent DEET. DEET was mixed in the diet and administered to CD rats for two years at concentrations that corresponded to dosage levels of 10, 30 or 100 mg/kg/day for males and 30, 100, or 400 mg/kg/day for females; to CD-1 mice for 18 months at dosage levels of 250, 500, or 1000 mg/kg/day; and to dogs for one year, via gelatin capsules, at dosage levels of 30, 100, or 400 mg/kg/day. In the rodent studies, each group consisted of 60 animals of each sex, and two concurrent independent control groups, each containing 60 animals/sex were included in each study. Each group in the dog study consisted of four male and four female dogs and one control group was included in the study. Treatment-related effects were observed at the highest dose level in all three studies. For rats, the effects included decreases in body weight and food consumption and an increase in serum cholesterol in females only. In mice, the effects observed were decreases in body weight and food consumption in both sexes. The effects observed in dogs included increased incidences of emesis and ptyalism, and levels of transient reduction in hemoglobin and hematocrit, increased alkaline phosphatase (males only), decreased cholesterol, and increased potassium. One male dog in the high-dose group also exhibited ataxia, tremors, abnormal head movements, and/or convulsions on several occasions during the study. The highest no-observed-effect levels (NO-ELs) for rats, mice and dogs were determined to be 100, 500, and 100 mg/kg/day, respectively. No specific target organ toxicity or oncogenicity was observed in any of the studies.

Acute and Subchronic Repeated Cutaneous Application of N-Methyldiethanolamine in the Fischer 344 Rat

The potential for local and systemic toxicity by recurrent contamination of the skin with N-methyldiethanolamine (MDEA; CAS No. 105-59-9) was investigated in the Fischer 344 rat. Two short-term repeated studies (9 days, 6 h/day) and one subchronic (65 applications over 13 weeks) were conducted with monitoring for clinical signs, body weight, food and water consumption, hematologic findings, clinical chemistry, urinalysis, gross and microscopic pathology analysis. The first 9 day study with undiluted MDEA (260,1040, and 2080 mg/kg/ day) produced dose-related skin irritation with related hematological and clinical chemistry changes. Adrenal weights were increased, a common finding in cutaneous irritation studies, and kidney weights were increased but without urinalysis or histological evidence of renal injury. Histopathologic investigation was limited to the skin treatment area. The second 9 day study, using aqueous dilutions of MDEA (100, 500, and 750 mg/kg/day) showed local irritation at the mid and high doses with associated clinical chemistry changes and increased adrenal gland weight. There were slight reductions in body weight gain. There were no effects on hematology or urinalysis, and histopathologic findings were limited to the treated skin. The subchronic study, also with aqueous dilutions (100, 250, and 750 mg/kg/day), showed dose-related irritation in the mid and high dose groups; major histopathological features were acanthosis, hyperkeratosis, parakeratosis, dermatitis, dermal fibrosis, eschar, and ulceration. Low-dose group findings were similar to the controls, and probably a consequence of the wrapping procedure. There were no effects on clinical pathological findings or organ weights, and histopathological findings were limited to treated skin. These studies show that recurrent application of MDEA produces a dose-related irritant effect, but there is no evidence for systemic cumulative or specific target organ or tissue toxicity.

Some milestones in in vitro organ Toxicity Assessment. The Kidney as a Case Study

The expression of target organ toxicity ranges from subtle abnormalities of cellular organelles to permanent loss of organ function. The selective targeting of chemicals for discrete regions and cell types of a given organ is frequently due (besides some pharmacodynamic mechanisms) to the fact that target cells may express unique biochemical or functional characteristics predisposing them to chemically induced injury. In vitro models commonly used in target organ toxicity tests include perfused organ preparations, isolated tissue preparations, single-cell suspensions and tissue culture systems. Although these systems have proved their usefulness for acute toxicity tests, there is still a great need for in vitro models to be used for chronic toxicity tests. Among the systems listed above, the single-cell culture technique may be adapted to long-term study requirements. The example of kidney proximal tubules is taken to illustrate the necessity for extensive characterization of the actual capacities of the models in term of phenotypic profiling, energy status, drug detoxication activities, specific transport systems and organ-specific differentiated functions. LLC-PK1, LLC-RK1, NRK and OK cell lines are compared with primary cultures of rat, rabbit and human proximal tubule cells. The importance of the cell culture environment on the cell phenotypic profile, and its subsequent response pattern to toxicant exposure, are described using gentamicin and platinum derivatives as examples. In terms of experimental strategy, choice of cell type, choice of species of origin, choice of doses, choice of duration, continuous or discontinuous exposure, and whether to study the recovery phase, are crucial issues for designing models mimicking more closely the in vivo situation. The identification of relevant endpoints, allowing discrimination between general cell toxicity and specific organ toxicity, has not been sufficiently explored in vitro. Scientifically based endpoints referring to the background studies conducted by biochemists or physiologists should be selected and included in experimental designs dealing with organ toxicology in vitro. Conceptually, relevant specific target-organ toxicity could be investigated by the use of multiple cell types and by analysis of the difference in concentration between the cytotoxic concentration threshold and the specific endpoint alteration threshold.