Systemic Toxicological Effects Research Articles

Diagnosis and Management of Paraquat Intoxication

Paraquat poisoning is a clinical toxicological emergency due to the active ingredient of this type of herbicide, gramoxone, with a high mortality rate due to high toxicity, and no antidote has yet been found. Paraquat intoxication can cause multi-organ failure if ingested accidentally or spontaneously because paraquat quickly produces ROS (reactive oxygen species), which causes cell damage through lipid peroxidation, NF-kB activation, mitochondrial damage, and apoptosis in many organs. This results in rapid nephrotoxic and hepatotoxic as well as pulmonary fibrosis. Clinical manifestations depend on the level of paraquat ingested, which can be in the form of local toxicological and systemic toxicological effects. Laboratory tests for diagnosing paraquat toxicity can be used for toxicological analysis of plasma and urine. Management of paraquat intoxication is primarily to remove paraquat from the gastrointestinal tract (prevent absorption) by using activated charcoal, changing the toxicokinetics of the herbicide (increasing serum paraquat elimination) by hemoperfusion and hemodialysis, as well as modifying its toxicodynamics with anti-inflammatory drugs such as immunosuppressants (corticosteroids and cyclophosphamide) and antioxidants (N -acetylcysteine and vitamin C).

Biodistribution and toxicology evaluation of a recombinant measles Schwarz-based Lassa vaccine in cynomolgus macaques.

MV-LASV is an investigational measles Schwarz-based vaccine for the prevention of Lassa fever. A repeated-dose toxicity study in cynomolgus macaques was performed to assess the biodistribution and local and systemic toxicological effects. Monkeys received three immunizations of MV-LASV or saline intramuscularly with a 2-week interval. An increase in anti-measles antibodies confirmed the reaction of the immune system to the vaccine backbone. Clinical observations, body weight, body temperature, local tolerance, electrocardiogram parameters, various clinical pathology parameters (hematology, coagulation urinalysis, serum chemistry, and C-reactive protein) were monitored. Gross pathology and histopathology of various tissues were evaluated. MV-LASV induced a mild increase in fibrinogen and C-reactive protein concentrations. This coincided with microscopic inflammation at the injection sites which partially or fully resolved following a 3-week recovery period. Viral RNA was found in secondary lymphoid organs and injection sites and gall bladder. No viral shedding to the environment was observed. Overall, the vaccine was locally and systemically well tolerated, supporting a first-in-human study.

Oral toxicity of titanium dioxide P25 at repeated dose 28-day and 90-day in rats

BackgroundNanotechnology is indispensable to many different applications. Although nanoparticles have been widely used in, for example, cosmetics, sunscreen, food packaging, and medications, they may pose human safety risks associated with nanotoxicity. Thus, toxicity testing of nanoparticles is essential to assess the relative health risks associated with consumer exposure.MethodsIn this study, we identified the NOAEL (no observed adverse effect level) of the agglomerated/aggregated TiO2 P25 (approximately 180 nm) administered at repeated doses to Sprague-Dawley (SD) rats for 28 and 90 days. Ten of the 15 animals were necropsied for toxicity evaluation after the repeated-dose 90-day study, and the remaining five animals were allowed to recover for 28 days. The agglomerated/aggregated TiO2 P25 dose levels used included 250 mg kg− 1 d− 1 (low), 500 mg kg− 1 d− 1 (medium), and 1000 mg kg− 1 d− 1 (high), and their effects were compared with those of the vehicle control. During the treatment period, the animals were observed for mortality, clinical signs (detailed daily and weekly clinical observations), functional observation battery, weekly body weight, and food and water consumption and were also subjected to ophthalmological examination and urinalysis. After termination of the repeated-dose 28-day, 90-day, and recovery studies, clinical pathology (hematology, blood coagulation time, and serum biochemistry), necropsy (organ weights and gross findings), and histopathological examinations were performed.ResultsNo systemic toxicological effects were associated with the agglomerated/aggregated TiO2 P25 during the repeated-dose 28-day, 90-day, and recovery studies in SD rats. Therefore, the NOAEL of the agglomerated/aggregated TiO2 P25 was identified as 1000 mg kg− 1 d− 1, and the substance was not detected in the target organs.ConclusionSubacute and subchronic oral administration of the agglomerated/aggregated TiO2 P25 was unlikely to cause side effects or toxic reactions in rats.

Use of Biocompatible Sorafenib-gold Nanoconjugates for Reversal of Drug Resistance in Human Hepatoblatoma Cells

The present study identifies the potential of highly biocompatible SF-GNP nano-conjugate to enhance the chemotherapeutic response to combat drug resistance in cancer cells. We developed a stable colloidal suspension of sorafenib-gold nanoconjugate (SF-GNP) of <10 nm size in aqueous medium for reverting the cancer drug resistance in SF-resistant HepG2 cells in a 3D ex-vivo model system. In-vivo biocompatibility assay of SF-GNPs showed absence of systemic toxicological effects including hematological, biochemical and histological parameters. More importantly, the histopathological analysis of vital organs such as liver, brain, lung, kidney and heart showed very least or no sign of inflammation, cell infiltration, necrosis, tissue disorganization or fibrotic reactions after intra-peritoneal administration of SF-GNP nanoconjugates in animals. However, SF-GNP nanoconjugates significantly reduced (>80%) the percentage cell survival and the size and number of SF resistant solid tumor colonies of HepG2 cells in 3D model system. The exposure of SF-GNP nanoconjugate to SF resistant HepG2 cell colonies also provided evidence for anti-proliferative effect and reversal of drug resistance by elucidating the molecular regulatory mechanisms of extracellular matrix factor (CD147), tumor growth factor (TGF-β), hepatoma upregulated protein (hURP) and drug transporter (ABCG-2).

Safety evaluation of cricket(Gryllus bimaculatus) extract in Sprague-Dawley rats

Recently, research investment in the improvement of food safety as a food source and specializing of nutritional source of edible insects is being actively conducted. Cricket especially has been attracting considerable interest in entomophagy; however, research on the safety assessment of cricket is limited. This study investigated the effects of cricket ethanol extract when orally administrated in Sprague-Dawley rats. Here, we performed a 4 wk repeated oral dose toxicity test in Sprague-Dawley rats following the Organization for Economic Cooperation and Development test guidelines 407 under Good Laboratory Practice regulation. Rats were randomly allocated 4 groups: vehicle control, 250, 500, 1,000 mg/kg test groups and administrated based on body weight for 28 d. The animals were observed for mortalities and clinical signs, body weight changes, food and water consumption. At the end of treatment period, blood and urine were collected and analyzed. Subsequently, the animals were sacrificed and subjected to gross pathological examination and organ weight measurement. The organs were preserved for histopathological examination. The results showed that there were no systemic toxicological effects related with the cricket ethanol extract in the 4 wk oral repeated dose toxicity study. It is considered that NOAEL of cricket ethanol extract is greater than 1,000 mg/kg/d and there was no target organ detected.

Assessment of inhaled acute ammonia-induced lung injury in rats

This study examined acute toxicity and lung injury following inhalation exposure to ammonia. Male Sprague-Dawley rats (300–350 g) were exposed to 9000, 20 000, 23 000, 26 000, 30 000 or 35 000 ppm of ammonia for 20 min in a custom head-out exposure system. The exposure atmosphere, which attained steady state within 3 min for all ammonia concentrations, was monitored and verified using a Fourier transform infrared spectroscopy (FTIR) gas analyzer. Animals exposed to ammonia resulted in dose-dependent increases in observed signs of intoxication, including increased chewing and licking, ocular irritation, salivation, lacrimation, oronasal secretion and labored breathing. The LCt50 of ammonia within this head-out inhalation exposure model was determined by probit analysis to be 23 672 ppm (16 489 mg/m3) for the 20 min exposure in male rats. Exposure to 20 000 or 23 000 ppm of ammonia resulted in significant body weight loss 24-h post-exposure. Lung edema increased in all ammonia-exposed animal groups and was significant following exposure to 9000 ppm. Bronchoalveolar fluid (BALF) protein concentrations significantly increased following exposure to 20 000 or 23 000 ppm of ammonia in comparison to controls. BAL cell (BALC) death and total cell counts increased in animals exposed to 20 000 or 23 000 ppm of ammonia in comparison to controls. Differential cell counts of white blood cells, neutrophils and platelets from blood and BALF were significantly increased following exposure to 23 000 ppm of ammonia. The following studies describe the validation of a head-out inhalation exposure model for the determination of acute ammonia-induced toxicity; this model will be used for the development and evaluation of potential therapies that provide protection against respiratory and systemic toxicological effects.

Hypoglycemic and High Dosage Effects of &amp;lt;i&amp;gt;Bidens pilosa&amp;lt;/i&amp;gt; in Type-1 Diabetes Mellitus

Ethno-pharmaceutical products have received a lot of international attention in the scientific community in the management of diabetes mellitus (DM). In this study we determined the anti-diabetic and high dosage effects of Bidens pliosa in type 1 DM (T1DM). Methodology: Thirty rats were divided into six groups and subgrouped into the extract and non extract treatment groups. The extract treated group was subdivided into three groups which received 200 mg/kg, 400 mg/ kg and 800 mg/kg dosage treatments respectively. The blood glucose levels were monitored using a standard glucometer for one month, and biochemical analysis of the two liver function enzymes; Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) were carried out at the Institute of Biomedical Research (IBR-KIU-WC) at the end of week IV. The study revealed that Bidens pilosa maintained hypoglycemia for a period of two weeks and this status was lost in subsequent weeks. T1DM rats treated with a dosage of 200 mg/kg showed a better recovery (355.25 - 164.5 mg/dl) of the glucose levels, followed by those that were being treated at 400 mg/kg. The AST and ALT enzymes in blood varied with a mean ± SEM (33.72 ± 32.32 to -7.23 ± 12.61 IU and 22.98 ± 11.12 to 42 ± 38.2 IU, respectively) in both the glibencimide? and in the 800 mg/ kg treatment groups in the study. High dosages of extract were associated (P = 0.049) with increased systemic enzyme leakage. In conclusion, tissue degeneration caused by high levels of the extract was accompanied by leakage of various enzymes (AST and ALT) into the blood, which could be a major etiological factor for the development of secondary systemic pathologies, thus potentially worsening the effects of an existing T1DM prognosis in human patients. The preliminary results indicate that a dose of Bidens pilosa has an anti-diabetic effect for a limited initial duration before starting to cause systemic toxicological effects. It is highly recommended that further investigation into the cellular mechanisms and consequences of any therapy involving Bidens pilosa be carried out.

The Targeted Histone Deacetylase Inhibitor Tefinostat (CHR-2845) Shows Selective In Vitro Efficacy In Monocytoid-Lineage Acute Myeloid Leukaemia (AML)

Tefinostat (CHR-2845) is a novel monocyte/macrophage-targeted histone deacetylase inhibitor (HDACi) that is cleaved to an active acid, CHR-2847, by an intracellular esterase (human carboxylesterase-1, hCE-1), found only in cells of monocytoid lineage and hepatocytes. The clinical uptake of HDAC inhibition to date has been restricted by systemic toxicities including gastrointestinal disturbance, thrombocytopenia and fatigue. Accumulation of CHR-2847 in hCE-1-expressing cells results in a 20-100-fold increase in targeted anti-proliferative potency, considerably widening the potential therapeutic window in malignancies involving cells of monocytoid lineage (AML-M4, AML-M5 and CMML) by sparing the systemic toxicological effects associated with non-selective HDAC inhibition.The in vitro efficacy of tefinostat was assessed in primary AMLs using stored mononuclear cells obtained at diagnosis from 70 AML patients. Dose-dependent induction of apoptosis and significant growth inhibitory effects were seen in M4 /M5 AMLs (median IC50; 1.1µM+/-1.8) compared to non-M4/M5 FAB types (median IC50 5.1µM +/-4.7) (p=0.007). This potency and monocytoid specificity was not reproduced when using an alternative HDACi, tefinostat analogue CHR-8185 which is not cleaved by hCE-1.hCE-1 protein expression in patient samples was measured by both intracellular flow cytometry and immunoblotting, with highest levels seen in M4/M5 patients. This observation was validated by microarray analysis of hCE-1 mRNA in a further 130 AML samples with M4/M5 AMLs showing significant overexpression compared to normal bone marrow CD34+ cells (p=0.009). High levels of hCE-1 expression were found to drive a significant increase in tefinostat efficacy as measured by growth inhibition assays (p=0.001), and also strongly correlated with expression of the mature monocytoid marker CD14+.Sub-population analysis by flow cytometry revealed variable sensitivity to tefinostat within AML blasts, with CD14+ expressing cells showing maximum growth inhibition. This CD14+ response was accompanied by an induction of intracellular protein acetylation at nanomolar concentrations in tefinostat-responsive samples. Tefinostat-sensitive samples also showed strong induction of the cell cycle arrest and DNA damage sensor protein pH2AX, which is a potential biomarker of patient responsiveness.Importantly, no growth inhibitory effects were seen in normal bone marrow cells (n=5) exposed to AML-toxic doses of tefinostat while, in comparison, equivalent concentrations of the non-hCE-1-dependent analogue CHR-8185 caused considerable cytotoxicity, again emphasising the potential for expansion of the clinical therapeutic window using an hCE-1-dependent agent.In vitro synergy was demonstrated in combination experiments with tefinostat and cytarabine (median Combination Index value=0.68) which is likely to be a logical combination for future clinical evaluation.In summary, monocytoid targeting of HDACi activity was achieved using tefinostat in primary AML samples of monocytoid lineage, with minimal toxicity to normal bone marrow cells at equimolar concentrations. Given the absence of significant toxicity seen in a recently-published phase 1 study of tefinostat in patients with advanced haematological malignancies, further larger scale clinical evaluation of this compound is warranted in haematological malignancies involving cells of monocytoid lineage. Disclosures:Zabkiewicz:Chroma Therapeutics: Research Funding. Gilmour:Chroma Therapeutics: Research Funding. Hills:Chroma Therapeutics: Research Funding. Bone:Chroma Therapeutics: Employment. Davidson:Chroma Therapeutics: Employment. Burnett:Chroma Therapeutics: Research Funding. Knapper:Chroma Therapeutics: Research Funding.

Tissue Distribution of Inhaled Micro- and Nano-sized Cerium Oxide Particles in Rats: Results From a 28-Day Exposure Study

In order to obtain more insight into the tissue distribution, accumulation, and elimination of cerium oxide nanoparticles after inhalation exposure, blood and tissue kinetics were investigated during and after a 28-day inhalation study in rats with micro- and nanocerium oxide particles (nominal primary particle size: < 5000, 40, and 5-10 nm). Powder aerosolization resulted in comparable mass median aerodynamic diameter (1.40, 1.17, and 1.02 μm). After single exposure, approximately 10% of the inhaled dose was measured in lung tissue, as was also estimated by a multiple path particle dosimetry model (MPPD). Though small differences in pulmonary deposition efficiencies of cerium oxide were observed, no consistent differences in pulmonary deposition between the micro- and nanoparticles were observed. Each cerium oxide sample was also distributed to tissues other than lung after a single 6-h exposure, such as liver, kidney, and spleen and also brain, testis, and epididymis. No clear particle size-dependent effect on extrapulmonary tissue distribution was observed. Repeated exposure to cerium oxide resulted in significant accumulation of the particles in the (extra)pulmonary tissues. In addition, tissue clearance was shown to be slow, and, overall, insignificant amounts of cerium oxide were eliminated from the body at 48- to 72-h post-exposure. In conclusion, no clear effect of the primary particle size or surface area on pulmonary deposition and extrapulmonary tissue distribution could be demonstrated. This is most likely explained by similar aerodynamic diameter of the cerium oxide particles in air because of the formation of aggregates and irrespective possible differences in surface characteristics. The implications of the accumulation of cerium oxide particles for systemic toxicological effects after repeated chronic exposure via ambient air are significant and require further exploration.

Biocompatibility of Dental Amalgams

Objective. The purpose of this review paper is to review the literature regarding the toxicology of mercury from dental amalgam and evaluate current statements on dental amalgam. Materials and Methods. Two key-words “dental amalgam” and “toxicity” were used to search publications on dental amalgam biocompatibility published in peer-reviewed journals written in English. Manual search was also conducted. The most recent declarations and statements were evaluated using information available on the internet. Case reports were excluded from the study. Results. The literature show that mercury released from dental amalgam restorations does not contribute to systemic disease or systemic toxicological effects. No significant effects on the immune system have been demonstrated with the amounts of mercury released from dental amalgam restorations. Only very rarely have there been reported allergic reactions to mercury from amalgam restorations. No evidence supports a relationship between mercury released from dental amalgam and neurological diseases. Almost all of the declarations accessed by the internet stated by official organizations concluded that current data are not sufficient to relate various complaints and mercury release from dental amalgam. Conclusions. Available scientific data do not justify the discontinuation of amalgam use from dental practice or replacement with alternative restorative dental materials.

Dermal Permeation of Biocides and Aromatic Chemicals in Three Generic Formulations of Metalworking Fluids

Metalworking fluids (MWF) are complex mixtures consisting of a variety of components and additives. A lack of scientific data exists regarding the dermal permeation of its components, particularly biocides. The aim of this study was to evaluate the dermal permeation of biocides and other aromatic chemicals in water and in three generic soluble oil, semi-synthetic, and synthetic MWF types in order to evaluate any differences in their permeation profiles. An in vitro flow-through diffusion cell study was performed to determine dermal permeation. An infinite dose of different groups of chemicals (6 biocides and 29 aromatic chemicals) was applied to porcine skin, with perfusate samples being collected over an 8-h period. Perfusate samples were analyzed by gas chromatography/mass spectrometry (GC-MS) and ultra-performance liquid chromatography/mass spectroscopy (UPLC-MS), and permeability was calculated from the analysis of the permeated chemical concentration–time profile. In general, the permeation of chemicals was highest in aqueous solution, followed by synthetic, semi-synthetic, and soluble oil MWF. The absorption profiles of most of the chemicals including six biocides were statistically different among the synthetic and soluble oil MWF formulations, with reduced permeation occurring in oily formulations. Permeation of almost all chemicals was statistically different between aqueous and three MWF formulation types. Data from this study show that permeation of chemicals is higher in a generic synthetic MWF when compared to a soluble oil MWF. This indicates that a soluble oil MWF may be safer than a synthetic MWF in regard to dermal permeation of chemicals to allow for an increased potential of systemic toxicity. Therefore, one may conclude that a synthetic type of formulation has more potential to produce contact dermatitis and induce systemic toxicological effects. The dilution of these MWF formulations with water may increase dermal permeability of biocides, allowing for an enhanced risk for systemic toxicological effects and dermatitis potential.

Chronic Toxicity and Reversibility of Antifertility Effect of Immunization against Gonadotropin-Releasing Hormone in Male Rats and Rabbits

The chronic systemic toxicity of immunization with gonadotropin-releasing hormone, conjugated to tetanus toxoid (GnRH-TT), was investigated in male rats and rabbits in order to start Phase I clinical trials. Groups of rats and rabbits were immunized with GnRH-TT dissolved in aqueous adjuvant. The antigen was administered at weeks 0, 4, and 8, followed by boosters to maintain high antibody titers. At termination (8-9 months after first immunization), twenty rats and ten rabbits exhibiting the highest mean anti-GnRH titers and all the controls were selected for complete toxicological evaluation. In the rat study, a castrated control group was included for comparison with the immunized group. The hematological and serum chemistry parameters of immunized rats and rabbits were not affected in a significant manner. Most of the changes in serum chemistry of immunized rats were also found in castrated rats, indicating that the changes are most likely due to the withdrawal of androgenic support. The weights of the testes, epididymides, and sex accessory glands were lower in all immunized animals. There was significant atrophy of the germinal epithelium, which, however, sustained a population of Sertoli cells, spermatogonia, and pachytene spermatocytes. Other morphological changes in the prostate, seminal vesicles, pituitary, and mammary gland reflected the effect of androgen withdrawal. The decrease in the weight of liver, kidney, and heart seen in the immunized rats was also present in castrated rats and was not associated with any histopathological changes. The reversibility of immunization-induced infertility was investigated by mating the rats with normal females. Four months after the start of immunization, 9 out of 10 immunized rats were infertile whereas by nine months, all rats had regained fertility. Thus, it is concluded that immunization with GnRH-TT had no systemic toxicological effects in the adult male rats and rabbits for the period studied. The results also indicated that the GnRH-TT immunization had an antifertility effect in male rats. Fertility was restored following cessation of immunization and decline in anti-GnRH antibody titers.

Metals and arsenic in eye shadows

The regulations relating to cosmetic products give no limit values for toxic elements such as metals or arsenic occurring as impurities in cosmetic products. The present study of metals (lead, cobalt, nickel, chromium) and arsenic in eye shadows in 88 colors of 25 brands and 49 products provides a basis for assessing the safety of eye shadow. 66 out of 88 (75%) of the colors contained more than 5 ppm of at least one of the elements, and all 49 products contained more than one 1 ppm of at least 1 of the elements. In one color, the amount of all elements was less than 1 ppm. The highest levels of cobalt and nickel were 41 and 49 ppm, respectively. These levels were high enough to cause allergic symptoms in those previously sensitized. Furthermore, long-term exposure to such levels may probably sensitize. The concentrations of arsenic were extremely low, 2.3 ppm at most. The level of lead was less than 20 ppm in all products. Accordingly, the concentrations of arsenic and lead seemed to be safe. 9 colors had concentrations of water-soluble chromium exceeding 2 ppm, and a very high level, 318 ppm, was encountered in 1 case. The overall results indicate that eye-shadows probably have no significant systemic toxicological effects. The groups at greatest risk are those already sensitized to the allergenic elements. Such consumers will have difficulties in choosing suitable products, since these elements in the form of impurities are not declared in the list of ingredients. Manufacturers should demand certification that the raw materials they buy contain no toxic elements. Although some of the products studied were acceptable, many had excessive levels of the elements from the consumer's viewpoint.

A 90-day subcutaneous toxicity and fertility study of a LHRH antagonist in rats

[Ac-D2Nal 1,4Cl-DPhe 2,D3Pal 3,Arg 5,DGlu 6 (anisole adduct),DAla 10]-GnRH (Nal-Glu) is an antagonist of LHRH and has the potential to be utilized as an antigonadal agent. A study was undertaken to evaluate the toxicological effects of Nal-Glu in rats. Nal-Glu, dissolved in 5% mannitol in water containing 9 ml/liter benzyl alcohol, was administered subcutaneously. In subchronic studies, groups of 12 male and 12 female rats received 0, 50, 250, or 1250 μg/kg body weight (BW) Nal-Glu for 90 days and were killed on Day 91. Additional groups of male and female rats were given the high dose of Nal-Glu (1250 μg/kg BW) or vehicle for either 30 or 90 days. Their fertility was assessed by mating them with normal animals. Unlike some other LHRH antagonists, Nal-Glu exhibited a low potency for causing in vitro histamine release from rat peritoneal mast cells. Furthermore, in acute in vivo studies, Nal-Glu was less active in the induction of peripheral edema. In the subchronic study, all doses of Nal-Glu were well tolerated and there were no apparent systemic toxic effects. The pharmacological effects of Nal-Glu were quite evident, however. Nal-Glu treatment led to a significantly decreased body weight gain in the males and a significantly increased body weight gain in the females. There was a dose-dependent decrease in weights of gonads and reproductive organs in both the sexes. Some of the hematological and serological parameters were significantly different in Nal-Glu-treated animals. However, most of the values were within the normal range and are considered to be of no toxicological significance. Histopathological evaluations were made in the control and high-dose groups only. In the male, a seminiferous tubular degeneration and atrophy of the interstitial cells was seen. The prostate and seminal vesicles were also atrophied and the epididymides were devoid of spermatozoa. In the females, the ovaries and uteri were atrophic. The injection site of Nal-Glu-treated rats had inflammatory changes indicative of a local irritating action of the drug. All other tissues had normal histomorphology. Both male and female rats became infertile when 1250 μg/kg Nal-Glu was administered for 30 days. Normal fertility was restored 8 weeks after cessation of 90-day treatment. It is concluded that repeated administration of Nal-Glu leads to reversible infertility in both male and female rats. Although it was irritating at the site of injection, Nal-Glu had no systemic toxicological effects.

Update on the Toxicity of Inhaled Methyl Methacrylate Vapor

The current threshold limit value (TLV) of 100 ppm for methyl methacrylate vapor was originally set because it was considered to be a value sufficiently low to preclude either systemic toxicological effects or discomfort from irritation in workers chronically exposed to the vapor. Shortly after this limit was proposed we became interested in evaluating the toxicological properties of methyl methacrylate vapor and, subsequently, of providing experimental data which would be useful in evaluating the adequacy of this TLV. The purpose of this article is to review some of the more relevant literature prior to 1976 which pertains to inhalation exposure and to summarize our more recent data which resulted from statistically designed studies employing acute and subchronic exposures to known concentrations of the vapor including the near-TLV value of 116 ppm.