Teratological Effects Research Articles

Cholinesterases (ChEs) and the cholinergic system in ontogenesis and phylogenesis, and non-classical roles of cholinesterases—A review

The enigma of the cholinergic function concerns the role of ChEs and other components of the cholinergic system in non-transmittive, non-synaptic phenomena. The notion that such unorthodox, non-classical phenomena must exist is clearly supported by several lines of evidence, such as the presence of ChEs and other cholinergic components early before neurogenesis, and indeed in unfertilized and fertilized eggs and in the sperm of many species, and their presence throughout phylogenesis, including non-motile, monocellular organisms, fungi and plants and many anervous and ephemeral tissues. The “flexibility” of ChEs, expressed in their polymorphism and their changeability during ontogenesis also speaks for the notion of non-classical functions of ChEs. Today, there is direct evidence that such functions do indeed exist, as for example, the evidence as to the role of ChEs and other cholinergic components in processes of cell proliferation and differentiation of synaptic and myoneural structures. Also, ChEs participate in cell communications as examplified by immunity processes, as well as pathological states, including Alzheimer's disease and states induced by “insults” such as stress and exposure to agents such as antiChEs. Finally, consistent with the non-classical roles of ChEs and cholinergic components are the morphogenetic and teratologic effects of antiChEs, including OP compounds and cholinergic agonists and antagonists. The structural homology between ChEs on the one hand, and adhesion molecules and protohormones on the other may explain some of this phenomenology. It is proposed that the phylogenetic ubiquity of ChEs and their basic capacities that are important for evolutionary phylogenesis, such as the capacity to promote cell adhesion and cell communication speaks for ChEs as “Ur” proteins.

Developmental and peri-postnatal study in cynomolgus monkeys with belimumab, a monoclonal antibody directed against B-lymphocyte stimulator

Belimumab is a fully human monoclonal antibody antagonist for soluble B-lymphocyte stimulator, and is a potential therapeutic for various autoimmune disorders. To support clinical use, belimumab was administered intravenously to pregnant cynomolgus monkeys every 2 weeks throughout gestation at dosages of 5 and 150 mg/kg. Fetuses were delivered by C-section on Gestation Day 150 from one-half of the mothers, and evaluated for teratologic effects (external, visceral, skeletal, and heart), pharmacodynamics (PD) and toxicokinetics (TK). Remaining mothers delivered their infants naturally, enabling extensive assessment of PD and TK during a 1-year postnatal period. Effects attributed to belimumab were limited to the expected pharmacology, primarily decreased numbers of B-lymphocytes in peripheral blood of mothers and infants, and in fetal lymphoid tissues. Infants demonstrated full recovery upon cessation of exposure. In conclusion, belimumab was well tolerated at pharmacologically active dose levels in pregnant cynomolgus monkeys and their infants after exposure throughout pregnancy.

Teratological Effects of Gamma-Irradiation during three Gestaional Intervals in Rats

Teratological Effects of Gamma-Irradiation during three Gestaional Intervals in Rats

Teratological Effects of Gamma-Irradiation during three Gestaional Intervals in Rats

Aim of the work: The present study aimed to evaluate the teratological effects of gamma-irradiation during three intervals of gestation; pre-implantation, organogenesis and fetal periods in rats. Material and methods: Four groups of pregnant rats were used in this study; the first one (GI) served as control. The second (GII), third (GIII) and fourth (GIV) groups were subjected to whole body γ-radiation at a sub-lethal single dose level of 4 Gy at the third day, 10th day and 14th day of gestation respectively.Pregnant rats were sacrificed at the 20th day of gestation, implantation sites, resorption, embryonic death, fetal death, growth retarded fetuses, external malformations and skeletal malformation were recorded. Results: The results showed that whole body γ-irradiation caused resorption in the embryos of pregnant rats especially in those exposed during the pre-implantation period than the two other periods. The embryonic and fetal deaths were prominent in the fetuses maternally exposed to whole body γ-irradiation during the organogenesis period. The highest percentage of growth retarded fetuses was found in fetuses maternally exposed to γ-rays during the organogenesis period followed by fetuses maternally exposed to γ-irradiation during the fetal period and then fetuses maternally exposed to γ-irradiation during the pre-implantation period. The skeletal malformations as a result of γ-irradiation were mostly represented in less ossification in the skull bones, less ossification in the vertebral centra and wavy ribs. The most affected fetal skeleton was perceptive in GIII (fetuses maternally exposed to γ-irradiation during the organogenesis period) followed by GII and then GIV.

Moderate use of alcohol, tobacco and cannabis during pregnancy: New approaches and update on research findings

Interest in fetal origins of adverse offspring outcomes has grown extensively in the last decade. This has resulted in many published studies focusing on exposure in utero to substances and human offspring outcomes. Exposure to maternal substance use in pregnancy is believed to be a preventable hazard, and is therefore a main issue for public health concern and policy. However, an important question in human studies remains whether prenatal substance use exposure has an aetiological role in pathways to adverse developmental and behavioural outcomes via teratological effects. Recent insights and developments in research methodology will aid the adequate and more refined testing of associations between prenatal substance use and offspring outcomes. In particular, novel approaches could assist in disentangling the exposure to substance effects from correlated risk factors. The purpose of this manuscript is therefore to provide an overview of methodological issues involved in studies that focus on the association between maternal substance use during pregnancy and offspring's outcomes, to describe novel approaches to test these associations, and present some examples of new and well-designed studies and discuss their findings.

Developmental consequences of prenatal tobacco exposure.

This paper reviews results from published, in press, and conference proceedings from 2007 and 2008 that link in-utero tobacco exposure to neurodevelopmental outcomes in exposed offspring. Prenatal tobacco exposure (PTE) affected speech processing, levels of irritability and hypertonicity, attention levels, ability to self-regulate, need to be handled, and response to novelty preference in infants. In early childhood, PTE effects were mostly behavioral outcomes including activity and inattention and externalizing behaviors, including conduct disorder and antisocial behavior. In adolescents, PTE predicted increased attention deficit hyperactivity disorder, modulation of the cerebral cortex and white matter structure, and nicotine addiction. Several studies found moderating effects with PTE and genetic susceptibilities including dopamine transporter, serotonergic synaptic function, and monomine oxidase pathways. Other studies suggested that environmental and genetic factors might be more important than the direct teratological effects of PTE. The majority of studies reviewed were prospective and tobacco exposure was quantified biologically. Most demonstrated a direct association between PTE and neurodevelopmental outcomes. More work is needed to examine multifactorial influences. Effects of PTE on the offspring appear to be moderated by genetic variability, neurobehavioral disinhibition, and sex.

Alterations in rat fetal morphology following abuse patterns of toluene exposure

Toluene is a commonly abused organic solvent. Inhalant abusers are increasingly women in their prime childbearing years. Children born to mothers who abused solvents during pregnancy may exhibit characteristics of a “fetal solvent syndrome” which may include dysmorphic features. This study examined the teratological effects of an abuse pattern of binge toluene exposure during gestation on skeletal and soft tissue abnormalities, body weight, and body size in fetal rats. Pregnant Sprague–Dawley rats were exposed for 30 min, twice daily, from gestational day (GD) 8 through GD20 to either air (0 ppm), 8000 ppm, 12,000 ppm, or 16,000 ppm toluene. Two-thirds of each litter was prepared for skeletal examination using Alizarin Red S staining while the remaining third of each litter was fixed in Bouin's solution for Wilson's soft tissue evaluation. Exposure to toluene at all levels significantly reduced growth, including decreases in placental weight, fetal weight, and crown-rump length. In addition, numerous gross morphological anomalies were observed such as short or missing digits and missing limbs. Skeletal examination revealed that ossification of the extremities was significantly reduced as a result of toluene exposure at all levels. Specific skeletal defects included misshapen scapula, missing and supernumerary vertebrae and ribs, and fused digits. Soft tissue anomalies were also observed at all toluene levels and there was a dose-dependent increase in the number of anomalies which included cryptorchidism, displaced abdominal organs, gastromegaly, distended/hypoplastic bladder, and delayed cardiac development, among others. These results indicate that animals exposed prenatally to levels and patterns of toluene typical of inhalant abuse are at increased risk for skeletal and soft tissue abnormalities.

Concentration and Persistence of Tin in Rat Brain and Blood Following Dibutyltin Exposure During Development

Dibutyltin (DBT), a widely used plastic stabilizer, has been detected in the environment as well as human tissues. Although teratological and developmental effects are well documented, there are no published reports of DBT effects on the developing nervous system. As part of a developmental neurotoxicity study of DBT, tissue samples were periodically collected to determine the distribution of total tin (Sn) in brain and whole blood. Pregnant Sprague-Dawley rats were exposed to 0, 10, or 25 ppm DBT in drinking water from gestational day (GD) 6 to weaning at postnatal day (PND) 21. Beginning on PND 3, half of the litters were directly dosed every 2 to 3 d via oral gavage with 0, 1, or 2.5 mg/kg DBT such that the dose level matched the water concentration (for example, litters with 25 ppm DBT in the water received 2.5 mg/kg). For Sn analysis, brain and blood samples were collected from culled pups on PND2 (males and females pooled), from pups (males and females separately) as well as dams at weaning (PND21), and from adult offspring (males and females) at PND93. Total Sn was quantified using inductively coupled plasma–mass spectroscopy (ICP-MS). At all ages, brain Sn levels were higher than blood. At culling, in the directly dosed pups at weaning, and in dams at weaning, Sn levels in both tissues were linearly related to dose. Weanling pups without direct dosing showed lower levels than either culled pups or dams, indicating that lactational exposure was minimal or negligible even while maternal exposure is ongoing. In the adults, Sn levels persisted in brains of directly dosed rats, and the high-dose females had higher levels than did high-dose males. No Sn was detected in adult blood. Thus, during maternal exposure to DBT in drinking water, Sn is placentally transferred to the offspring, but lactational transfer is minimal, if any. Furthermore, Sn is concentrated in brain compared to blood, and its elimination is protracted, on the order of days to months after exposure ends.

The teratologic effect of Vincristine during organogenesis in newborn mice

The teratologic effect of Vincristine during organogenesis in newborn mice

병아리 태자에 대한 엔로플록사신의 최기형 독성 연구

The purpose of this study was to investigate the teratological effects of enrofloxacin, a veterinary antibiotic, on the embryos and fetus of hatching chicken eggs. A control group and four experimental fertilized egg groups were set up. The four experimental groups were injected with 0.05 ml, 0.1 ml, 0.2 ml, and 0.3 ml enrofloxacin (<TEX>$50{\mu}g/ml$</TEX>) respectively. During the hatching period, the weights of total eggs, inside material(yolk and white) and embryos or fetuses, embryo growth, and teratological effects were investigated. In the experimental groups (fertilized eggs injected with 0.05 ml, 0.1 ml, 0.2 ml and 0.3 ml enrofloxacin), the weights of total eggs were decreased, but the yolk and white weights were increased by the higher amount of antibiotics. The weights of embryos (fetuses) in experimental groups were 86.1%, 78.6%, 65.6% and 61.4% of the control, respectively. Retarded growth, deformity and embryo loss were observed in experimental groups. Teratological effects such as undeveloped eyes, wings and legs, and deformed head and bill were also detected. In conclusion, we found that veterinary antibiotics, enrofloxacin have made teratological effects on the embryos and fetus of hatching eggs.

Meaningful differences in maternal smoking behaviour during pregnancy: implications for infant behavioural vulnerability

Background:Smoking during pregnancy has been consistently associated with risk of problem behaviour in offspring. There is debate about whether this association reflects a teratological effect or is a marker for...

Prenatal Cocaine Exposure: Drug and Environmental Effects at 9 Years

To assess school-age cognitive and achievement outcomes in children with prenatal cocaine exposure, controlling for confounding drug and environmental factors. At age 9 years, 371 children (192 cocaine exposure [CE]; 179 non-cocaine exposure [NCE]) were assessed for IQ and school achievement in a longitudinal, prospective study from birth. An extensive number of confounding variables were controlled, including quality of caregiving environment, polydrug exposure, blood lead level, iron-deficiency anemia (IDA), and foster/adoptive care. Prenatal cocaine exposure predicted poorer perceptual reasoning IQ, with a linear relationship of the concentration of the cocaine metabolite benzoylecgonine to the degree of impairment. Effects were mediated through birth head circumference, indicating a relationship with fetal brain growth. Negative effects of alcohol, lead, and marijuana exposure and positive effects of the home environment were additive. The CE children in foster/adoptive care had better home environments and lower lead levels. School achievement was not affected. Persistent teratologic effects of CE on specific cognitive functions and additive effects of alcohol, lead, and marijuana exposure; IDA; and the home environment were identified. Documenting environmental factors in behavioral teratology studies is important, because in this sample, CE was associated with better home environment and lower environmental risk in a substantial number of children.

Quantification of Teratological Effects of Infectious Bronchitis Virus Isolated from Commercial Poultry in Pakistan

2 Abstract: In continuation of our studies with the Infectious Bronchitis prevalence in Pakistan, the current study was conducted to determine the teratological effects of the Infectious Bronchitis Virus (IBV) on developing chicken embryo. The tissues collected (Trachea and Lungs) from clinically positive IB chickens were tested with the indirect immunofluorescence assay against M-41 IBV strain (reported previously). The positive samples were further tested with the RT-PCR against the IBV primers (reported previously). The tissue homogenates from 43 such RT-PCR-positive samples were serially passaged through 11 days old embryos and the effects on the embryonic development in terms of dwarfing, curling, stunting and urates deposits were recorded. The data showed that the passaging sequentially increased the IBV teratological effects. Only 13 out of 43 samples exhibited IBV-associated teratological effects over various passages. These effects were effectively neutralizes by using IBV variant specific antisera in a viral neutralization test. These studies imply that IBV variants isolated from Pakistan are no different in producing teratological effects considered representative of the IBV.

The oxidizing enzyme CYP26a1 tightly regulates the availability of retinoic acid in the gastrulating mouse embryo to ensure proper head development and vasculogenesis

Retinoic acid (RA) has been implicated as one of the signals providing a posterior character to the developing vertebrate central nervous system. Embryonic RA first appears in the posterior region of the gastrulating embryo up to the node level, where it may signal within the adjacent epiblast and/or newly induced neural plate to induce a hindbrain and spinal cord fate. Conversely, rostral head development requires forebrain-inducing signals produced by the anterior visceral endoderm and/or prechordal mesoderm, and there is evidence that RA receptors must be in an unliganded state to ensure proper head development. As RA is a diffusible lipophilic molecule, some mechanism(s) must therefore have evolved to prevent activation of RA targets in anterior regions of the embryo. This might result from RA catabolism mediated by the CYP26A1 oxidizing enzyme, which is transiently expressed in anteriormost embryonic tissues; however, previous analysis of Cyp26a1(-/-) mouse mutants did not clearly support this hypothesis. Here we show that Cyp26a1(-/-) null mutants undergo head truncations when exposed to maternally-derived RA, at doses that do not affect wild-type head development. These anomalies are linked to a widespread ectopic RA signaling activity in rostral head tissues of CYP26A1-deficient embryos. Thus, CYP26A1 is required in the anterior region of the gastrulating mouse embryo to prevent teratological effects that may result from RA signaling. We also report a novel role of CYP26A1 during early development of the intra- and extra-embryonic vascular networks.

Modeling the relationship of cotinine and self-reported measures of maternal smoking during pregnancy: A deterministic approach

Studies of effects of prenatal exposure to cigarettes frequently acquire both self-report and biological assays of maternal smoking. However, little attention has been paid to methods for combining information from both sources to enhance the precision of exposure measurement. This paper analyzes the relationship between the two commonly used measures of smoking exposure during pregnancy: Maternal self-report and urinary cotinine. We present a deterministic method for combining the two measures and examine its robustness under different assumptions. We apply the method to a dataset from the Family Health and Development Project. In addition, we propose an approach for calibrating the self-report measures for individual women based on both sources of information. Enhancing the quality of exposure measurement may substantially advance studies of the teratological effects of exposure on offspring.

First Cell Cycles of Sea Urchin Paracentrotus lividus Are Dramatically Impaired by Exposure to Extremely Low-Frequency Electromagnetic Field

Exposure of fertilized eggs of the sea urchin Paracentrotus lividus to an electromagnetic field of 75-Hz frequency and low amplitudes (from 0.75 to 2.20 mT of magnetic component) leads to a dramatic loss of synchronization of the first cell cycle, with formation of anomalous embryos linked to irregular separation of chromatids during the mitotic events. Because acetylcholinesterase (ACHE) is thought to regulate the embryonic first developmental events of the sea urchin, its enzymatic activity was assayed in embryo homogenates and decreased by 48% when the homogenates were exposed to the same pulsed field. This enzymatic inactivation had a threshold of about 0.75 +/- 0.01 mT. The same field threshold was found for the effect on the formation of anomalous embryos of P. lividus. Moreover, ACHE inhibitors seem to induce the same teratological effects as those caused by the field, while blockers of acetylcholine (ACh) receptors are able to antagonize those effects. We conclude that one of the main causes of these dramatic effects on the early development of the sea urchin by field exposure could be the accumulation of ACh due to ACHE inactivation. The crucial role of the membrane in determining the conditions for enzyme inactivation is discussed.

Amended Final Report on the Safety Assessment of Oryza Sativa (Rice) Bran Oil, Oryza Sativa (Rice) Germ Oil, Rice Bran Acid, Oryza Sativa (Rice) Bran Wax, Hydrogenated Rice Bran Wax, Oryza Sativa (Rice) Bran Extract, Oryza Sativa (Rice) Extract, Oryza Sativa (Rice) Germ Powder, Oryza Sativa (Rice) Starch, Oryza Sativa (Rice) Bran, Hydrolyzed Rice Bran Extract Hydrolyzed Rice Bran Protein,

This report addresses the safety of cosmetic ingredients derived from rice, Oryza sativa. Oils, Fatty Acids, and Waxes: Rice Bran Oil functions in cosmetics as a conditioning agent--occlusive in 39 formulations across a wide range of product types. Rice Germ Oil is a skin-conditioning agent--occlusive in six formulations in only four product categories. Rice Bran Acid is described as a surfactant-cleansing agent, but was not in current use. Rice Bran Wax is a skin-conditioning agent--occlusive in eight formulations in five product categories. Industry did not directly report any use of Rice Bran Wax. Hydrogenated Rice Bran Wax is a binder, skin-conditioning agent--occlusive, and viscosity-increasing agent--nonaqueous in 11 formulations in six product categories. Rice Bran Oil had an oral LD50 of > 5 g/kg in white rats and Rice Wax had an oral LD50 of > 24 g/kg in male mice. A three-generation oral dosing study reported no toxic or teratologic effects in albino rats fed 10% Rice Bran Oil compared to a control group fed Peanut Oil. Undiluted Rice Bran Oil, Rice Germ Oil, and Hydrogenated Rice Bran Wax were not irritants in animal skin tests. Rice Bran Oil was not a sensitizer. Rice Bran Oil, Rice Germ Oil, Rice Wax, and Hydrogenated Rice Bran Wax were negative in ocular toxicity assays. A mixture of Rice Bran Oil and Rice Germ Oil had a ultraviolet (UV) absorption maximum at 315 nm, but was not phototoxic in a dermal exposure assay. Rice Bran Oil was negative in an Ames assay, and a component, gamma-oryzanol, was negative in bacterial and mammalian mutagenicity assays. Rice oils, fatty acids, and waxes were, at most, mildly irritating in clinical studies. Extracts: Rice Bran Extract is used in six formulations in four product categories. Rice Extract is a hair-conditioning agent, but was not in current use. Hydrolyzed Rice Extract is used in four formulations and current concentration of use data were provided for other uses. Hydrolyzed Rice Bran Extract, described as a skin-conditioning agent--miscellaneous, is used in two product categories. Use concentrations are in the 1% to 2% range. Rice Bran Extract is comprised of proteins, lipids, carbohydrates, mineral ash, and water. The content includes palmitic, stearic, oleic, and linoleic acids. Other components include antioxidants such as tocopherols. Rice Extract reduced the cytotoxicity of sodium chloride in male rats. Bran, Starch and Powder: Rice Bran (identified as rice hulls) is an abrasive and bulking agent in one formulation. Rice Starch is an absorbent and bulking agent in 51 formulations across a wide range of product categories. Rice Germ Powder is an abrasive and one manufacturer described an exfoliant use, but it was not reported to be used in 2002. Oral carcinogenicity studies done on components of Rice Bran (phytic acid and gamma-oryzanol) were negative. Rice Bran did not have an anticarcinogenic effect on 1,2-dimethylhydrazine-induced large bowel tumors. In cocarcinogenicity studies done using 1,2-dimethylhydrazine and other agents, with Rice Bran Oil and Rice Bran-derived hemicellulose and saccharide, tumor inhibition was observed; gamma-oryzanol did not inhibit the development of neoplasms. A decrease in cutaneous lesions in atopic dermatitis patients was reported following bathing with a Rice Bran preparation. Proteins: Hydrolyzed Rice Bran Protein and Hydrolyzed Rice Protein function as conditioning agents (hair or skin), but only the latter was reported to be used in a few products. An in vitro phototoxicity assay using UVA light found no photochemical toxicity. Rice bran protein hydrolysates are not acutely toxic, are not skin or ocular irritants in animals, are not skin sensitizers in guinea pig maximization tests, and are not irritating or sensitizing in clinical tests. Isolated cases of allergy to raw rice have been reported, but rice, in general, is considered non allergenic. The Cosmetic Ingredient Review (CIR) Expert Panel considered that safety test data available on certain of these ingredients could be extrapolated to the entire group. Although Rice Bran Extract does contain UV absorbing compounds at low concentrations, clinical experience suggested no phototoxicity would be associated with such materials. Rice derived ingredients generally are considered to be non allergenic. There were no safety test data available for Hydrolyzed Rice Extract and Hydrolyzed Rice Bran Extract, but their safety may be inferred from that of the extracts from which they are derived. Current levels of polychlorinated biphenyls (PCBs) and heavy metals in rice-derived ingredients used in cosmetics are not a safety concern. The Panel was concerned, however, that contaminants such as pesticides have been reported in Rice Bran Oil used for cooking. Pesticides and heavy metals should not exceed currently reported levels for rice-derived cosmetic ingredients. The CIR Expert Panel concluded that these rice-derived ingredients are safe as cosmetic ingredients in the practices of use and concentrations as described in this safety assessment.

Differential teratogenesis of all‐trans‐retinoic acid administered on gestational day 9.5 to SWV and C57BL/6N mice: Emphasis on limb dysmorphology

Mouse strain differences in teratologic response are well documented. However, because retinoids cause similar malformation syndromes across many species, the strain differences may be predicted to be minimal. The goals of this study were to characterize and explain the differences between the C57BL/6N and SWV mouse strains in terms of all-trans-retinoic acid (RA)-induced teratologic effects at the time of gestation that cause postaxial forelimb ectrodactyly. Visceral and skeletal malformations were determined by Wilson's sectioning and double-staining techniques, respectively; developmental staging was performed according to the somite count; and retinoid concentrations were assessed by HPLC. C57BL/6N mice were more susceptible than SWV mice to induction of embryolethality, cardiovascular defects, and forelimb ectrodactyly, whereas the opposite was true for the induction of ear, thymus, and tail agenesis, and cleft palate, gastroschisis, and anal atresia. As determined by somite counts, 1 strain intercross was developmentally advanced compared to the parental strains and the reciprocal cross. Retinoid susceptibility was equivalent between the reciprocal crosses for some malformations and determined by the maternal genotype for others. Toxicokinetic experiments showed that whole-embryo peak retinoid concentrations did not differ between the strains, but the area under the curve (AUC) for all-trans-RA was 1.3 times higher in C57BL/6N than in SWV embryos. The malformation spectrum induced by RA was strain-specific, and the strain sensitivity for forelimb ectrodactyly was consistent with all previously tested teratogenic agents (i.e., C57BL/6N was more sensitive than SWV). The strain differences in teratologic effects were not explained by developmental timing differences or toxicokinetic differences at the whole-embryo level.

Attenuating effects of natural organic matter on microcystin toxicity in zebra fish (Danio rerio) embryos—benefits and costs of microcystin detoxication

To contribute to the understanding of joined factors in the environment, impact of pure microcystins (-RR and -LF) on zebra fish (Danio rerio) embryos were investigated individually and in combination with a natural organic matter (NOM). The applied NOM was a reverse osmosis isolate from Lake Schwarzer See (i.e., Black Lake, BL-NOM). Teratogenic effects were evaluated through changes in embryonic development within 48 h of exposure. Detoxication activities were assessed by the activities of phase II biotransformation enzymes, soluble and microsomal glutathione S-transferase (s, mGST). Oxidative stress was assessed by determining both the production of hydrogen peroxide and by analyzing the activities of the antioxidative enzymes, guajacol peroxidase (POD), catalase (CAT), glutathione peroxidase (GPx), and the glutathione restoring enzyme glutathione reductase (GR). Energetic costs were evaluated by determining contents of fat, carbohydrates, and proteins in both exposed and control embryos. BL-NOM attenuated toxic effects of MC-LF and MC-RR verified by less pronounced teratological effects within 24 h, in particular, as well as less rise in the activity of s-GST, when compared with embryos exposed to either pure toxins or in combination with organic matter. BL-NOM also diminished oxidative effects caused by MC-LF; however, it failed to attenuate oxidative stress caused by MC-RR. Content of lipids was significantly reduced in exposed embryos following a trend similar to that obtained with teratological and enzymatic assays confirming the attenuating effect of BL-NOM. Physiological responses to microcystins and NOM required energetic costs, which were compensated to the expense of the energy resources of the yolk, which in turn might affect the normal development of embryos.

Studies of the Toxicological Potential of Tripeptides (L-Valyl-L-prolyl-L-proline and L-lsoleucyl-L-prolyl-L-proline): VI. Effects of Lactobacillus helveticus-fermented Milk Powder on Fertility and Reproductive Performance of Rats

The objective of these studies was to assess the effects of the tripeptides, L-valyl-L-prolyl-L-proline (VPP) and L-isoleucyl-L-prolyl-L-proline (IPP), on reproductive capabilities of male and female rats. The specific goals of the experiments were (1) to determine the effects of orally administered tripeptides on (a) fertility and reproductive behavior in both sexes of rats, (b) embryo-fetal development in pregnant rats, and (c) pre- and postnatal development of rats exposed to tripeptides in utero and during lactation; and (2) to estimate the no-observable-adverse-effect doses of tripeptides in maternal and fetal rats. During the conduct of these classical segment I, II, and III studies, the test material was powdered Lactobacillus helveticus-fermented milk (FM), which contains the tripeptides, VPP and IPP. FM (0, 500, 1000 or 2000 mg/kg body weight [BW]/day--equivalent to 0, 0.8, 1.6, or 3.3 mg/kg BW/day of VPP plus IPP) was administered to males by oral gavage from 4 weeks prior to mating until sacrifice, and to females from 2 weeks prior to mating through day 20 of lactation. Evaluative parameters included monitoring grossly observable clinical signs; food consumption and body weight gains; mating behavior and fertility indices of both sexes; implantation and maintenance of embryos; sex ratio of live pups; fetal viability; incidences of external, visceral or skeletal variations; growth and behavioral development; as well as reproductive capabilities of F1 offspring exposed to FM during gestation and lactation. All animals were subjected to macroscopic examination at termination of their segment of the studies. Clinical signs, body weights, and food consumption were unaffected by administration of FM. During segment I, the test agent had no effect on estrus cycle, mating behavior, fertility index, or reproductive competence of either males or females. The results of segment II experiments revealed no effects of FM on postimplantation survival-loss, sex ratio or birth weights of live fetuses, and there was no evidence of treatment-associated developmental or teratological effects. During segment III, FM was without effect on pup viability, behavioral and sexual maturation, and reproductive capability of the F1 generation. Under the conditions of these experiments, the no-observable-adverse-effect level (NOAEL) of FM on reproductive performance in male and female rats is greater than 2000 mg/kg BW/day, the equivalent of 3.3 mg/kg BW/day of VPP plus IPP.