Short-term Toxicity Studies Research Articles

Safety Assessment of Poloxamers 101, 105, 108, 122, 123, 124, 181, 182, 183, 184, 185, 188, 212, 215, 217, 231, 234, 235, 237, 238, 282, 284, 288, 331, 333, 334, 335, 338, 401, 402, 403, and 407, Poloxamer 105 Benzoate, and Poloxamer 182 Dibenzoate as Used in Cosmetics

Poloxamers are polyoxyethlyene, polyoxypropylene block polymers. The impurities of commercial grade Poloxamer 188, as an example, include low-molecular-weight substances (aldehydes and both formic and acetic acids), as well as 1,4-dioxane and residual ethylene oxide and propylene oxide. Most Poloxamers function in cosmetics as surfactants, emulsifying agents, cleansing agents, and/or solubilizing agents, and are used in 141 cosmetic products at concentrations from 0.005% to 20%. Poloxamers injected intravenously in animals are rapidly excreted in the urine, with some accumulation in lung, liver, brain, and kidney tissue. In humans, the plasma concentration of Poloxamer 188 (given intravenously) reached a maximum at 1 h, then reached a steady state. Poloxamers generally were ineffective in wound healing, but were effective in reducing postsurgical adhesions in several test systems. Poloxamers can cause hypercholesterolemia and hypertriglyceridemia in animals, but overall, they are relatively nontoxic to animals, with LD(50) values reported from 5 to 34.6 g/kg. Short-term intravenous doses up to 4 g/kg of Poloxamer 108 produced no change in body weights, but did result in diffuse hepatocellular vacuolization, renal tubular dilation in kidneys, and dose-dependent vacuolization of epithelial cells in the proximal convoluted tubules. A short-term inhalation toxicity study of Poloxamer 101 at 97 mg/m(3) identified slight alveolitis after 2 weeks of exposure, which subsided in the 2-week postexposure observation period. A short-term dermal toxicity study of Poloxamer 184 in rabbits at doses up to 1000 mg/kg produced slight erythema and slight intradermal inflammatory response on histological examination, but no dose-dependent body weight, hematology, blood chemistry, or organ weight changes. A 6-month feeding study in rats and dogs of Poloxamer 188 at exposures up to 5% in the diet produced no adverse effects. Likewise, Poloxamer 331 (tested up to 0.5 g/kg day(-1)), Poloxamer 235 (tested up to 1.0 g/kg day(-1)), and Poloxamer 338 (at 0.2 or 1.0 g/kg day(-1)) produced no adverse effects in dogs. Poloxamer 338 (at 5.0 g/kg day(-1)) produced slight transient diarrhea in dogs. Poloxamer 188 at levels up to 7.5% in diet given to rats in a 2-year feeding study produced diarrhea at 5% and 7.5% levels, a small decrease in growth at the 7.5% level, but no change in survival. Doses up to 0.5 mg/kg day(-1) for 2 years using rats produced yellow discoloration of the serum, high serum alkaline phosphatase activity, and elevated serum glutamicpyruvic transaminase and glutamic-oxalacetic transaminase activities. Poloxamers are minimal ocular irritants, but are not dermal irritants or sensitizers in animals. Data on reproductive and developmental toxicity of Poloxamers were not found. An Ames test did not identify any mutagenic activity of Poloxamer 407, with or without metabolic activation. Several studies have suggested anticarcinogenic effects of Poloxamers. Poloxamers appear to increase the sensitivity to anticancer drugs of multidrug-resistant cancer cells. In clinical testing, Poloxamer 188 increased the hydration of feces when used in combination with a bulk laxative treatment. Compared to controls, one study of angioplasty patients receiving Poloxamer 188 found a reduced myocardial infarct size and a reduced incidence of reinfarction, with no evidence of toxicity, but two other studies found no effect. Poloxamer 188 given to patients suffering from sickle cell disease had decreased pain and decreased hospitilization, compared to controls. Clinical tests of dermal irritation and sensitization were uniformly negative. The Cosmetic Ingredient Review (CIR) Expert Panel stressed that the cosmetic industry should continue to use the necessary purification procedures to keep the levels below established limits for ethylene oxide, propylene oxide, and 1,4-dioxane. The Panel did note the absence of reproductive and developmental toxicity data, but, based on molecular weight and solubility, there should be little skin penetration and any penetration of the skin should be slow. Also, the available data demonstrate that Poloxamers that are introduced into the body via routes other than dermal exposure have a rapid clearance from the body, suggesting that there would be no risk of reproductive and/or developmental toxicity. Overall, the available data do not suggest any concern about carcinogenesis. Although there are gaps in knowledge about product use, the overall information available on the types of products in which these ingredients are used, and at what concentration, indicates a pattern of use. Based on these safety test data and the information that the manufacturing process can be controlled to limit unwanted impurities, the Panel concluded that these Poloxamers are safe as used.

Flavouring Group Evaluation 5, Revision 1 (FGE.05Rev1):Esters of branched‐ and straight‐chain aliphatic saturated primary alcohols and of one secondary alcohol, and branched‐ and straight‐chain unsaturated carboxylic acids from chemical groups 1, 2, and 5 (Commission Regulation (EC) No 1565/2000 of 18 July 2000) ‐ Opinion of the Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in contact with

Flavouring Group Evaluation 5, Revision 1 (FGE.05Rev1):Esters of branched‐ and straight‐chain aliphatic saturated primary alcohols and of one secondary alcohol, and branched‐ and straight‐chain unsaturated carboxylic acids from chemical groups 1, 2, and 5 (Commission Regulation (EC) No 1565/2000 of 18 July 2000) ‐ Opinion of the Scientific Panel on Food Additives, Flavourings, Processing Aids and Materials in contact with

Studies on cytotoxicity and antitumor screening of red and white forms of Abrus precatorius L.

The cytotoxicity study with cancer cell line A-549 indicated that the methanol insoluble fraction of crude red forms of Abrus precatorius is toxic (CTC50 175,100 mg/ml) in microculture tetrazolium assay and sulphorhodamine B assays, whereas crude and fractions of the red and white forms exhibited toxicity at still higher concentrations. However, an antitumor screening by the short-term toxicity study with DLA cells showed the extracts to be comparatively less toxic.

Evaluation of statistical tools used in short-term repeated dose administration toxicity studies with rodents

In order to know the different statistical tools used to analyze the data obtained from twenty-eight-day repeated dose oral toxicity studies with rodents and the impact of these statistical tools on interpretation of data obtained from the studies, study reports of 122 numbers of twenty-eight-day repeated dose oral toxicity studies conducted in rats were examined. It was found that both complex and easy routes of decision trees were followed for the analysis of the quantitative data. These tools include Scheffe's test, non-parametric type Dunnett's and Scheffe's tests with very low power. Few studies used the non-parametric Dunnett type test and Mann-Whitney's U test. Though Chi-square and Fisher's tests are widely used for analysis of qualitative data, their sensitivity to detect a treatment-related effect is questionable. Mann-Whitney's U test has better sensitivity to analyze qualitative data than the chi-square and Fisher's tests. We propose Dunnett's test for analysis of quantitative data obtained from twenty-eight-day repeated dose oral toxicity tests and for qualitative data, Mann-Whitney's U test. For both tests, one-sided test with p=0.05 may be applied.

Toxicity Profile of Lutein and Lutein Ester Isolated From Marigold Flowers (Tagetes erecta)

Lutein is a carotenoid with antioxidant properties and is commonly present in many fruits, vegetables, and egg yolk. Lutein affords protection against the development of the two common eye diseases of aging: cataract and macular degeneration. As the dietary lutein concentration is much lower compared to the actual requirement to reduce macular degeneration, supplementation of lutein is under consideration. There are very few data on the toxicity of lutein. In the present study, the authors have evaluated the short-term and long-term toxicity profile of lutein and its esterified form isolated from marigold flowers (Tagetes erecta) in young adult male and female Wistar rats. Lutein and its ester form administered orally at doses of 4, 40, and 400 mg/kg body weight for 4 weeks for short-term toxicity study and 13 weeks for a subchronic toxicity study did not produced any mortality, change in body weight, food consumption pattern, organ weight, and other adverse side reactions. Administration of lutein and ester form did not alter the hepatic and renal function, and did not produce any change in the hematological parameters and in lipid profile. Histopathological analysis of the organs supported the nontoxicity of lutein and its ester form.

Antitumor and antioxidant status of Galega purpurea root in Ehrlich ascites carcinoma bearing Swiss albino mice

The present study was designed to determine the antitumor and antioxidant properties of methanol extract from the root of Galega purpurea (Papilionaceae) (MEGP) against Ehrlich Ascites Carcinoma (EAC) bearing Swiss albino mice. Acute and short-term toxicity studies were performed initially in order to ascertain the safety of MEGP. The effect of MEGP on the growth of transplantable murine tumor, life span of EAC bearing hosts and simultaneous alterations in the hematological profile and liver biochemical parameters (lipid peroxidation, antioxidant enzymes) were estimated. The MEGP showed decrease in tumor volume, packed cell volume and viable cell count and increases the nonviable cell count and mean survival time thereby increasing life span of EAC tumor bearing mice. Hematological profile reverted to more or less normal levels in extract treated mice. Treatment with MEGP decreased the levels of lipid peroxidation and increased the levels of glutathione, superoxide dismutase and catalase. The results suggested that the methanol extract of Galega purpurea root exhibited antitumor effect by modulating lipid peroxidation and augmenting antioxidant defense system in EAC bearing mice.

Effects of Microdesmis keayana roots on sexual behavior of male rats

In the present study, the aphrodisiac properties of Microdesmis keayana J. Léonard root extract and major isolated alkaloids were evaluated by observing the sexual behavior of male rats. Aqueous extract (150mg/kg body weight) and pure alkaloids (3mg/kg body weight) were administered orally by gavage to male rats. Latent times of observation, intromission and ejaculation, mounting behavior, number of intromissions and mating performances were evaluated and compared to those obtained with untreated rats in the presence of receptive and non-receptive females. The results have shown that aqueous extract and alkaloids of M. keayana stimulate sexual parameters in rats' sexual behavior. A short-term toxicity study undertaken to establish the therapeutic index of aqueous extract, showed that a high dose of the extract (2g/kg body weight) caused no mortality or changes in rats' behavior.

Vasoactivity, antioxidant and aphrodisiac properties of Caesalpinia benthamiana roots

Aims of the study Caesalpinia benthamiana (=Mezo neuron benthamianum) (Fabaceae) is an African tropical plant whose roots are used in traditional medicine as an aqueous decoction for many purposes, especially for erection impairment but its action mechanism is unknown. The action of Caesalpinia benthamiana on sexual behaviour and some assays on potential modes of action were performed. Materials and Methods The aqueous extract of Caesalpinia benthamiana (AECB) was tested for vasorelaxing properties using isolated rat aortic rings precontracted by phenylephrine. Influence of AECB in the production of endothelial isoform of nitric oxide synthase (eNOS) was measured by quantitative polymerase chain reaction (QPCR) analysis. Scavenging activities versus reactive oxygen species (ROS), such as superoxide anion (O 2 −), hydrogen peroxide (H 2O 2), and hypochlorous acid (HOCl) were assessed. Action of AECB on the cellular generation of O 2 − was also tested in a physiopathology model of oxidative burst using human polymorphonuclear neutrophils stimulated by phorbol myristate acetate. The aphrodisiac properties of AECB administered orally by gavage (50 mg/kg body weight) to male rats were evaluated by observing the sexual behaviour of animals. Finally, a short-term toxicity study was undertaken to establish the therapeutic index of AECB administered orally to rats at high dose (2 g/kg body weight). Results C. benthamiana roots are rich in phenolic compounds (gallic acid, resveratrol and tannins). The results showed that AECB had significant vasorelaxing properties. The extract also had a strong radical activity against ROS in cell-free and cellular systems and stimulated eNOS mRNA expression. As for the aphrodisiac activity of AECB in male rats, results have shown that sexual parameters are stimulated. Furthermore, after oral administration at high dose, AECB causes no mortality or changes in rats’ behaviour. Conclusion AECB enhanced the sexual activity of male rats. This could be partly explained by its vasorelaxant properties, which may be caused by an increase in NO production in vascular bed and a decrease in its destruction.

Genomics and proteomics analysis of cultured primary rat hepatocytes

The use of animal models in pharmaceutical research is a costly and sometimes misleading method of generating toxicity data and hence predicting human safety. Therefore, in vitro test systems, such as primary rat hepatocytes, and the developing genomics and proteomics technologies, are playing an increasingly important role in toxicological research. Gene and protein expression analysis were investigated in a time series (up to 5 days) of primary rat hepatocytes cultured on collagen coated dishes. Especially after 24 h, a significant down-regulation of many important Phase I and Phase II enzymes (e.g., cytochrome P450’s, glutathione- S-transferases, sulfotransferases) involved in xenobiotic metabolism, and antioxidative enzymes (e.g., catalase, superoxide dismutase, glutathione peroxidase) was observed. Acute-phase-response enzymes were frequently up-regulated (e.g., LPS binding protein, α-2-macro-globulin, ferritin, serine proteinase inhibitor B, haptoglobin), which is likely to be a result of cellular stress caused by the cell isolation procedure (perfusion) itself. A parallel observation was the increased expression of several structural genes (e.g., β-actin, α-tubulin, vimentin), possibly caused by other proliferating cell types in the culture, such as fibroblasts or alternatively by hepatocyte dedifferentiation. In conclusion, the careful interpretation of data derived from this in vitro system indicates that primary hepatocytes can be successfully used for short-term toxicity studies up to 24 h. However, culturing conditions need to be further optimized to reduce the massive changes of gene and protein expression of long-term cultured hepatocytes to allow practical applications as a long-term toxicity test system.

E, E, E-1-(4-Arylamino-4-oxo-2-butenoyl)-3,5-bis(arylidene)-4-piperidones: A topographical study of some novel potent cytotoxins

A series of E, E, E-3,5-bis(arylidene)-1-(4-arylamino-4-oxo-2-butenoyl)-4-piperidones 4 (phenylidene) and 5 (4-nitrophenylidene) were prepared in order to explore the structural features of the N-acyl group which affects the cytotoxic potency. Evaluation toward human Molt 4/C8 and CEM T-lymphocytes revealed that many of the IC 50 figures were submicromolar and lower than melphalan. Marked inhibitory potencies toward murine leukemia L1210 cells were also noted. When evaluated against a panel of human tumor cell lines, three representative compounds in series 4 displayed selective toxicity to leukemia and colon cancer cell lines and were significantly more potent than the reference drug melphalan. Molecular modeling of representative compounds in both series 4 and the analogs, in which the configuration of the olefinic double bond was changed from E to Z (series 3), revealed that the torsion angles of the arylidene aryl rings and locations of the terminal arylaminocarbonyl groups may have contributed to the greater cytotoxic properties displayed in 3. Compounds 4c (3,4-dichlorophenylamino), d (4-methylphenylamino) and 5c (3,4-dichlorophenylamino), d (4-methylphenylamino) inhibited the activity of human N-myristoyltransferase by approximately 50% at concentrations of 50–100 μM. The compounds in series 4 and 5 were well tolerated in a short-term toxicity study in mice.

Final Report on the Safety Assessment of Capsicum Annuum Extract, Capsicum Annuum Fruit Extract, Capsicum Annuum Resin, Capsicum Annuum Fruit Powder, Capsicum Frutescens Fruit, Capsicum Frutescens Fruit Extract, Capsicum Frutescens Resin, and Capsaicin1

Final Report on the Safety Assessment of Capsicum Annuum Extract, Capsicum Annuum Fruit Extract, Capsicum Annuum Resin, Capsicum Annuum Fruit Powder, Capsicum Frutescens Fruit, Capsicum Frutescens Fruit Extract, Capsicum Frutescens Resin, and Capsaicin1

Safety studies on epigallocatechin gallate (EGCG) preparations. Part 2: Dermal, acute and short-term toxicity studies

Green tea extract and its principal active ingredient, epigallocatechin gallate (EGCG), are gaining attention and increased usage due to their healthful properties. Despite the increasing demand for these products, few studies have examined their safety. The toxicity of purified green tea extracts containing high concentrations of EGCG have been evaluated in a series of studies in order to define the safety of Teavigo™, a high-concentration EGCG extract produced by the same novel method. Topical EGCG preparations caused minor dermal irritation in rats and guinea pigs, but not rabbits, and was a moderate dermal sensitizing agent in the guinea pig maximization test. A rabbit eye irritation test produced a strong enough response to not warrant any further testing in this assay. An oral dose delivering 2000 mg EGCG preparation/kg was lethal to rats; whereas, a dose of 200 mg EGCG/kg induced no toxicity. The dietary administration of EGCG preparation to rats for 13 weeks was not toxic at doses up to 500 mg/kg/day. Similarly, no adverse effects were noted when 500 mg EGCG preparation/kg/day was administered to pre-fed dogs in divided doses. This dose caused morbidity when administered to fasted dogs as a single bolus dose, although this model was considered an unrealistic comparison to the human condition. From these studies a no-observed adverse effect level of 500 mg EGCG preparation/kg/day was established.

Deformities in cane toad (Bufo marinus) populations in Bermuda: Part II. Progress towards characterization of chemical stressors

An alarming high incidence of deformities in mature and newly metamorphosed Bufo marinus was identified in 2002 (29% in adults and 21% in metamorph specimens) in Bermuda. Suspicion of multiple chemical stressors in many of the effected ponds warranted identification of potential causative agents. Short-term embryo-larval, chronic exposure studies through hind limb development, and a battery of toxicity identification studies were performed with B. marinus and the surrogate species, Xenopus laevis, using sediment and water collected from four target ponds and one reference pond in Bermuda. Short-term embryo larval developmental toxicity studies based on a modified FETAX method indicated that samples from each of the target sites were capable of inducing abnormal development. Malformations induced in the early developmental tests were consistent amongst the species and included abnormal development of the craniofacial region, eye, mouth, and gut. Limb reduction deficit defects were identified in the chronic exposure studies with B. marinus and X. laevis. Toxicity characterization studies found that the toxicity of sediment extracts was variably filterable and reduced by C18-SPE, cation exchange, and zeolite treatments. Developmentally toxic samples contained elevated levels of petroleum hydrocarbons, metals (Al, As, Sn, Cd, Cr, Cu, Fe, Pb, Hg, Mn, Ni, and Zn), and ammonia. Sediment spiking studies with reference sediment confirmed that the levels of petroleum hydrocarbons and metals were capable of inducing developmental toxicity individually. Joint mixture interaction studies with sediment extracts prepared from the individually spiked reference sediment suggested however, that these extracts were potentially capable of acting synergistically, producing a greater level of toxicity than would normally be predicted based on the response to the individual spiked samples.

BENEFICIAL EFFECT OF MODERATE FOOD RESTRICTION IN TOXICITY STUDIES IN RATS

Moderate food restriction (FR) has been established as a nutritionally appropriate and well-controlled method with long-term beneficial effects in conducting toxicity and carcinogenicity studies in rodents. This study describes the early effects of moderate FR on toxicity study parameters in rats and on the variability of these parameters. Physical signs, body weight, food and water consumption, and clinical pathology parameters were examined in a 4-week study in which rats were moderately food-restricted or fed ad libitum (AL). There were no diet-related differences in physical signs, hematology or urinalysis. FR-related changes were observed in body weight and serum biochemistry; however, most of the changes involved anti-aging alterations and/or physiological adjustment to FR. Moderate FR resulted in low variability and good reproducibility in body weight. The present results indicate that moderate FR does not impair study parameters and increases statistical sensitivity. Therefore, a moderate FR feeding regimen is beneficial not only for long-term but also for short-term toxicity studies in rats.

랫드에서 고환독성의 정색을 위한 정량적 평가법의 확립: 2-bromopropane의 예

The aims of the study were to establish a short-term screening test for detecting testicular toxicity of chemicals in rats and to determine whether a 2-week administration period is sufficient to detect testicular toxicity of 2-bromopropane (2-BP) as an example. Male Sprague-Dawley rats were subcutaneously administered with 1000 mg/kg/day of 2-BP or its vehicle for 2 weeks. Ten male rats each were sacrificed on days 3, 7 and 14 after the initiation of treatment. Parameters of testicular toxicity included genital organ weights, testicular sperm head counts, epididymal sperm counts, motility and morphology, and qualitative and quantitative histopathologic examinations. The early histopathological changes observed on day 3 of treatment included degeneration of spermatogonia and spermatocytes, multinuclear giant cells, mature spermatid retention, vacuolization of Sertoli cells, and decreased number of spermatogonia in stages II and V. On day 7 of treatment, atrophy of seminiferous tubules, exfoliation of germ cells, degeneration of spermatogonia and spermatocytes, multinuclear giant cells, mature spermatid retention, vacuolization of Sertoli cells, decreased number of spermatogonia in stages II and V, and decreased number of spermatocytes in stages VII and XII. On day 14 after treatment, a significant decrease in the weights of testes and seminal vesicles was found. Atrophy of seminiferous tubules, exfoliation of germ cells, degeneration of spermatogonia and spermatocytes, mature spermatid retention, vacuolization of Sertoli cells, decreased number of spermatogonia in stages II and V, and decreased number of spermatocytes in all spermatogenic stages were also observed. In addition, a slight non-significant decrease in testicular sperm head counts, daily sperm production rate and epididymal sperm counts was found. The results showed that 2 weeks of treatment is sufficient to detect the adverse effects of 2-BP on male reproductive organs. It is considered that the short-term testicular toxicity study established in this study can be a useful tool for screening the testicular toxic potential of new drug candidates in rats.

Preliminary Report on the Absence of Toxicity of Fruit Pulp of Palmyrah (Borassus Flabellifer L.) In ICR Mice

Palmyrah (Borassus flabellifer L.) flour prepared from the seed shoot has been previously reported to cause hepatotoxic, neurotoxic and other toxic effects. Although palmyrah fruit pulp (PFP) is also consumed in Sri Lanka, no toxicity studies have been done. Short and long term toxicity studies were therefore conducted with PFP by administration of 10% or 50% PFP (dry basis) incorporated into WHO standard feed (part of the maize being substituted with PFP) compared with a control group of ICR mice, fed the WHO standard feed (n=7 in each group, age 6 weeks, weight 36- 44g) , for 7 or 30 days. In both studies the effect of 10% or 50% PFP feed on liver and kidney function and haematological parameters (haemoglobin concentration, packed cell volume, white blood cell count and red blood cell count) were determined. Liver function was assessed by estimation of serum levels of alanine transaminase, aspartate transaminase and alkaline phosphatase. Renal toxicity was assessed by estimation of serum creatinine levels. In the long term toxicity study, effect of PFP on histology of main body organs (heart, lung, liver, kidney and intestine) were also assessed by microscopic examination of haematoxylin / eosin stained sections of these organs. Results of both the short term and long term toxicity studies demonstrated that PFP at the 10% or 50% level did not have any significant effect on kidney or liver function nor on the haematological parameters under study and on the histology of major body organs in ICR mice. In all studies, neurotoxicity was assessed by comparing the behaviour ofthe test group with the control group. No behavioural changes or spasms were observed in the test groups of mice fed with the feed containing 10% or 50% PFP. The overall results suggest that PFP even at the 50% level in the standard feed probably does not produce any toxic effects in ICR mice.

A review of evidence from short-term studies leading to the prediction that diazoaminobenzene (1,3-diphenyltriazine) is a carcinogen

The National Toxicology Program (NTP) is responsible for providing comprehensive toxicology evaluations of substances, while at the same time incorporating approaches to reduce, refine or replace laboratory animals in routine toxicity/carcinogenicity studies. Consistent with this, a series of metabolism studies in rodents and human liver slices, electron spin resonance spectroscopy (ESR) studies, short-term dermal toxicity studies in rodents, and acute bone marrow micronucleus studies in mice were performed on diazoaminobenzene (DAAB, also known as 1,3-diphenyltriazine). These studies demonstrated that DAAB is metabolized and shares similar genotoxic and toxicological properties to the known human carcinogen, benzene, and the known rodent carcinogen, aniline. These data were used to evaluate the potential carcinogenicity of DAAB without doing a 2-year rodent bioassay. Based on this analysis, DAAB was predicted to be carcinogenic if evaluated in a 2-year rodent bioassay. These data were evaluated to support listing DAAB in the NTP Report on Carcinogens as a substance 'reasonably anticipated to be a human carcinogen'. The purpose of this article is to review the data developed for predicting the carcinogenicity of DAAB.

Two Cases of Adreno-Hepatic Fusion in Cynomolgus Monkeys (Macaca fascicularis)

In this report we describe 2 cases of adreno-hepatic fusion (AHF) in Cynomolgus monkeys (Macaca fascicularis) used in short-term toxicity studies. AHF is defined as the union of hepatic tissue with the adrenal gland with close intermingling of the respective parenchymal cells. In this condition, a continuous intervening connective tissue septum is lacking. AHF is believed to be a congenital anomaly caused by a defect or delay in the formation of the organ capsules from the intermediate primitive mesenchymal stroma. To our knowledge, this is the first time this anomaly is described in the monkey.

Constituents of aromatic plants: elemicin

No results of short-term or chronic toxicity studies have been found. Elemicin did induce UDS in hepatocytes from male rats. Studies on carcinogenicity were negative, but the 1′-hydroxy-metabolite of elemicin gave positive and negative results. The total intake of elemicin from essential oil seems to be limited. The main source of intake appears to be nutmeg. Further studies are needed to evaluate if the intake of elemicin may represent a health risk.

Short-term oral toxicity of homoyessotoxins, yessotoxin and okadaic acid in mice

A short-term toxicity study after 7 days oral daily administration of yessotoxin (YTX; 2 mg/kg/day), homoYTX (1 mg/kg/day), 45-hydroxy-homoYTX (1 mg/kg/day) and of the main diarrhoetic shellfish toxin okadaic acid (OA; 1 mg/kg/day) was carried out in mice. Symptoms, lethality, food consumption, body and organ weights, gross pathology and histopathology of the main organs and tissues, leukocytes formula as well as plasmatic levels of transaminases, lactate dehydrogenase and creatinine phosphokinase were evaluated. Heart tissue was studied also hystochemically for the presence of apoptotic nuclei and by transmission electron microscopy. No mortality, signs of toxicity or cumulative effects were induced by the repeated oral exposure to YTXs. Only ultrastructural changes in the cardiac muscle cells near the capillaries, such as package of rounded mitochondria and alteration of the cells boundary were observed, without any increase of lactate dehydrogenase, an index of cardiac damage. OA induced diarrhoea, body weight loss, reduced food consumption, and the death of 2/5 mice after 5 days. Necroscopy and/or light microscopy analysis revealed toxic effects mainly at forestomach (ulceration and hyperplasia), liver and, indirectly to body weight loss of mice, atrophic signs in the lymphoid organs and exocrine pancreas. Electron microscopy of heart tissue showed alterations of mitochondria and fibers in myocardiocytes, although no apoptotic change was recorded.