Toxicological Parameters of a Formulation Containing Cinnamaldehyde for Use in Treatment of Oral Fungal Infections: An In Vivo Study.

OBJECTIVE: We aimed to define the safety and toxicity of both isolated and embedded cinnamaldehyde using a pharmaceutical formulation for the treatment of oral fungal infections in an in vivo study. MATERIALS AND METHODS: Acute toxicity was assessed in studies with Galleria mellonella larvae and Danio rerio embryos (zebrafish), and genotoxicity was assessed in a mouse model. The pharmaceutical formulation (orabase ointment) containing cinnamaldehyde was evaluated for verification of both in vitro antifungal activity and toxicity in keratinized oral rat mucosa. RESULTS: In Galleria mellonella larvae, cinnamaldehyde was not toxic up to the highest dose tested (20 mg/kg) and presented no genotoxicity up to the dose of 4 mg/kg in the model using mice. However, it was found to be toxic in zebrafish embryos up to a concentration of 0.035 µg/mL; LC50 0.311; EC50 0.097 (egg hatching delay); and 0.105 (Pericardial edema). In the orabase antifungal susceptibility test, cinnamaldehyde exhibited activity in concentrations greater than 200 µg/mL. As for safety in the animal model with rats, the orabase ointment proved to be safe for use on keratinized mucosa up to the maximum concentration tested (700 µg/mL). CONCLUSIONS: At the concentrations tested, cinnamaldehyde was not toxic in vertebrate and invertebrate animal models and did not exhibit genotoxic activity. In addition, when used in the form of an ointment in orabase, having already recognized antifungal activity, it was shown to be safe up to the highest concentration tested.

Avaliação da toxicidade pré-natal: estudo de teratogenicidade do inseticida piriproxifeno em ratos Wistar / Prenatal toxicity assessment: teratogenicity study of the insecticide pyriproxyfen in Wistar rats

Este estudo investigou a toxicidade pré-natal do inseticida piriproxifeno em ratos Wistar, de forma a detectar possíveis alterações no desenvolvimento fetal da progênie exposta durante o período organogênico. Três doses de piriproxifeno (100, 300 e 500mg.kg-1) foram administradas por via oral às progenitoras, do sexto ao 15º dia de gestação. Os fetos foram submetidos à técnica de diafanização modificada descrita por Taylor e Van Dyke, para avaliação de malformações e alterações esqueléticas. Os resultados não demonstraram a ocorrência de toxicidade materna sistêmica ou alterações nos índices reprodutivos avaliados. Malformações ou alterações teratogênicas não foram detectadas, no entanto alterações esqueléticas sugestivas de retardo no desenvolvimento foram observadas especialmente nas doses mais altas testadas (300mg.kg-1 e 500mg.kg-1). Considerando-se a situação complexa de risco para a saúde humana, mostra-se importante a execução de investigações adicionais, de modo a contribuir para a adequada avaliação de risco do piriproxifeno em água potável.(AU)

This study investigated the prenatal toxicity of the insecticide pyriproxyfen in Wistar rats to detect the possible changes in the fetal development of the progeny exposed during the organogenic period. Three doses of pyriproxyfen (100, 300, and 500mg.kg-1) were administered orally to the progenitors, from day 6 to 15 of gestation. The fetuses were processed using the Taylor and Van Dyke modified diaphanization technique to evaluate malformations and skeletal changes. The results did not demonstrate the occurrence of systemic maternal toxicity or changes in the reproductive indexes evaluated. Malformations or teratogenic changes were not detected, however, skeletal changes suggestive of developmental delay were observed, especially in the highest doses tested (300 mg.kg-1 and 500 mg.kg-1). Owing to the potentially complex situation regarding its risk to human health, it is important that further studies be performed to contribute to the risk assessment of the addition of pyriproxyfen in drinking water.(AU)

Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization.

Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by in vitro fertilization (IVF). In this study, we conducted superovulation, IVF, and knock out and knock in using IVF rat embryos. We found that superovulation effectively occurred in the synchronized oestrus cycle and with anti-inhibin antiserum treatment in immature rats, including the Brown Norway rat, which is a very difficult rat strain to superovulate. Next, we collected superovulated oocytes under anaesthesia, and offspring derived from IVF embryos were obtained from all of the rat strains that we examined. When the tyrosinase gene was targeted by electroporation in these embryos, both alleles were disrupted with 100% efficiency. Furthermore, we conducted long DNA fragment knock in using adeno-associated virus and found that the knock-in litter was obtained with high efficiency (33.3-47.4%). Thus, in this study, we developed methods to allow the simple and efficient production of model rats.

CCN3 secreted by prostaglandin E2 inhibits intimal cushion formation in the rat ductus arteriosus.

The ductus arteriosus (DA), an essential fetal shunt between the pulmonary trunk and the descending aorta, changes its structure during development. Our previous studies have demonstrated that prostaglandin E2 (PGE2)-EP4 signaling promotes intimal cushion formation (ICF) by activating the migration of DA smooth muscle cells via the secretion of hyaluronan. We hypothesized that, in addition to hyaluronan, PGE2 may secrete other proteins that also regulate vascular remodeling in the DA. In order to detect PGE2 stimulation-secreted proteins, we found that CCN3 protein was increased in the culture supernatant in the presence of PGE2 in a dose-dependent manner by nano-flow liquid chromatography coupled with tandem mass spectrometry analysis and enzyme-linked immunosorbent assay. Quantitative RT-PCR analysis revealed that PGE2 stimulation tended to increase the expression levels of CCN3 mRNA in DA smooth muscle cells. Immunohistochemical analysis revealed that CCN3 was highly localized in the entire smooth muscle layers and the endothelium of the DA. Furthermore, exogenous CCN3 inhibited PGE2-induced ICF in the ex vivo DA tissues. These results suggest that CCN3 is a secreted protein of the DA smooth muscle cells induced by PGE2 to suppress ICF of the DA. The present study indicates that CCN3 could be a novel negative regulator of ICF in the DA to fine-tune the PGE2-mediated DA remodeling.

Rat Articular Cartilages Change Their Tissue and Protein Compositions During Perinatal Period.

Articular cartilage (AC) covers the surface of bones in joints and functions as a cushion against mechanical loading. The tissue contains abundant extracellular matrix (ECM), which mainly consists of proteoglycans (PG) and collagen (COL) fibres. The property of AC is gradually changing by ageing with gravity loading. To know the property change of AC by initial gravity loading during short period after birth, we performed histological assays and proteomics assay on the AC of the femoral condyle in knee joints of perinatal rats. The water content (%) was significantly decreased in neonate AC compared with fetal AC. During the perinatal stages (E19 and P0), the localizations of glycosaminoglycan (GAG) and type I and II COLs were homogeneous. The density of chondrocytes was significantly decreased in the deeper layers comparing with the surface layer in neonate AC. In addition, we found a drastic change in the protein expression pattern on proteomic analysis. The expressions of ECM components were relatively increased in neonate AC compared with fetal AC.

Influência do estrógeno e do interferon γ sobre a expressão da indoleamina-2,3-dioxigenase em cultura de células de placenta e embriões de ratas Wistar / Estrogen and interferon γ influence on 2, 3 indoleamine-dioxygenase expression in culture of placenta and fetal cells from Wistar rats

A indoleamina 2,3-dioxigenase (IDO) é uma enzima responsável por catabolizar o aminoácido triptofano. Sua presença no ambiente uterino placentário está relacionada à tolerância imunológica ao semi-aloenxerto, pois impede a proliferação de células imunológicas maternas, seja pela falta do aminoácido, ou pela ação de alguns catabólitos oriundos da quebra do triptofano, como o ácido quinolínico, que é tóxico principalmente para os linfócitos T. Pouco se conhece sob a influência de substâncias (hormônios e citocinas) presentes na interface materno fetal e a expressão dessa enzima. Por esta razão, formulou-se a hipótese de que hormônios e interleucinas presentes na região uteroplacentária poderiam exercer algum efeito na expressão da IDO. Células oriundas da interface materno fetal de ratas Wistar foram mantidas em cultivo, onde receberam suplementação com estradiol e interferon-γ. A expressão da enzima foi avaliada pela técnica de citometria de fluxo nos períodos de 4, 24 e 48 horas e confirmação da presença proteica por imuno-histoquímica. Os resultados mostraram um aumento na expressão de IDO após a adição de estrógeno (9,03±0,81/11,25±0,25) e interferon-γ (9,03±0,81/20,43±0,60). O efeito do interferon-γ já era esperado como relatado na literatura, contudo, a elevação da expressão da IDO pela adição do estrógeno constitui nova informação sobre possíveis mecanismos envolvidos na ativação da enzima. O melhor esclarecimento desses achados poderia contribuir para uma melhor compreensão da participação dessa enzima na tolerância materno-fetal e para uma futura modulação terapêutica da mesma...

The indoleamine 2,3-dioxygenase (IDO) is an enzyme responsible for catabolizing the tryptophan. Its presence in the placental uterine environment is related to immunological tolerance to the semi-allograft because it prevents proliferation of maternal immune cells, either by the lack of this amino acid or by the action of its catabolites, such as the quinolinic acid, which is particularly toxic for T lymphocytes. Little is known regarding the influence of hormones and cytokines on the expression of IDO in the maternal fetal interface. Therefore, the hypothesis that some hormones and interleukins present in uteroplacental region could have an effect on the expression of IDO on cultured cells was tested. Cells derived from the fetal maternal interface from Wistar rats were kept in culture and supplemented with estradiol and interferon-γ. Expression of the enzyme was assessed by flow cytometry at periods of 4, 24 and 48 hours and confirmation of the presence of protein by immunohistochemistry. The results showed an increasing of IDO expression after the addition of estrogen (9.03±0.81 to 11.25±0.25) and interferon-γ (9.03±0.81 to 20.43±0.60). The effect of interferon-γ was expected as reported in the literature, however, elevated IDO expression by estrogen represents new information on possible mechanisms involved in the enzyme activation. These findings could provide a better understanding of IDO contribution on maternal-fetal tolerance and may collaborate to future therapeutic modulation of this enzyme...

Efeitos teratogênicos de Prosopis juliflora em ratos e análise da toxicidade das vagens / Teratogenic effects of Prosopis juliflora in rats and analysis of the toxicity of the pods

Objetivou-se com este estudo determinar os possíveis efeitos teratogênicos de Prosopis juliflora, verificar se existe perda da toxicidade entre vagens armazenadas e recém-coletadas e determinar se existe diferença de toxicidade entre as vagens coletadas em diferentes localidades. Trinta ratas prenhes da linhagem Wistar foram separadas, aleatoriamente, em cinco grupos: um controle (G1) e quatro experimentais (G2, G3, G4 e G5). Os animais dos grupos G2 e G3 foram alimentados com ração contendo 70% de vagens de P. juliflora recém-coletadas em dois municípios diferentes. Os grupos G4 e G5 foram alimentados com ração preparada com vagens das mesmas procedências, porém armazenadas por um período de 6 meses. O grupo controle recebeu ração sem vagens de P. juliflora. No grupo controle o número de malformações por ninhada (1,16 ± 0,98) foi significativamente menor do que os dos grupos experimentais (14,00 ± 2,96, 6,16 ± 2,22, 7,66 ± 2,94 e 4,66 ± 1,63 para os grupos G2, G3, G4 e G5, respectivamente) indicando que a planta é teratogênica. Não foram observadas diferenças significativas na frequência de malformações e no número de fetos nascidos entre os grupos que receberam vagens de diferentes localidades. No entanto, o número de malformações nos grupos que receberam as vagens recém-colhidas (65,80 ± 21,20 a, 67,59 ± 18,10 a), foi significativamente maior que o observado nas ratas que receberam as vagens após o armazenamento (35,74±10,10b, 21,85±5,13c) sugerindo que o efeito teratogênico da planta diminui durante o armazenamento. Conclui-se que as vagens de P. juliflora são teratogênicas para ratas Wistar e que a fetoxicidade das mesmas diminui com o armazenamento.(AU)

The objective of this study was to determine the possible teratogenic effects of Prosopis juliflora, check if there is a loss in toxicity between pods stored and newly collected and determine whether there are differences in toxicity between the pods collected in different localities. Thirty pregnant female Wistar rats were randomly separated into five groups: a control (G1) and four experimental (G2, G3, G4 and G5), each with six animals. The animals in groups G2 and G3 were fed diets containing 70% of pods of P. juliflora newly collected in two different municipalities. The groups G4 and G5 were fed beans prepared with the same origins, but stored for a period of 6 months. The control group received the same diet without pods of P. juliflora. In the control group the number of defects per litter (1.16 ± 0.98) was significantly lower than the experimental groups (14.00 ± 2.96, 6.16 ± 2.22, 7.66 ± 2.94 and 4.66 ± 1.63 for G2, G3, G4 and G5, respectively) indicating that the plant is teratogenic. No significant differences were observed in the frequency of malformations and number of fetuses born between groups receiving pods from different locations. However, the number of defects in the groups who received the freshly harvested pods was significantly different from the number observed in rats that received the beans after storage, suggesting that the teratogenic effect of the plant decreases during storage. We conclude that the pods of P. juliflora are teratogenic for Wistar rats and that the teratogenicity decreases with storage.(AU)

[The combined effect of lead and zinc on the embryonic development of laboratory rats].

In the article there are presented the results of the study of the impact of inorganic lead and zinc compounds, as well as their organic forms produced with the use of nanotechnoloy, on the embryonic development of laboratory rats. Metals were orally administered daily during 19 days of gestation at the doses of 0.05 mg/kg of lead, and 1.5 mg/kg of zinc. The impact of the test substances was evaluated by integral and specific indices with the use of physiological, morphological and quantitative methods of analysis. Lead in a dose of 0.05 mg/kg was established to disturb the antenatal development of the offspring of experimental animals, which is pronounced in the increased embryo lethality rate, deterioration of somatometric indices of male fetuses in the litter as compared with the control group, and compared with females. In permits to suggest the greater sensitivity of male fetuses to exposure to lead. The isolated impact of zinc in the dose of 1.5 mg/kg body weight does not influence on the levels of embrio mortality rate, as well as somatometric indices of fetuses. However, the combined administration of the compounds of zinc and lead weakens the embryotoxic effect of the latter in terms of embrio lethality and the amount of live fetuses in the litter with more effective bioprotection for zinc in the nanoaquachelate form.

Alterações placentárias em resposta à exposição de ratas Wistar à poluição atmosférica / Placental alterations in response to exposure of Wistar rats to air pollution

Introdução: A exposição à poluição atmosférica durante a gestação provoca alterações nas características da placenta e pode resultar em restrição de crescimento intrauterino. Sabe-se que o transforming growth factor beta 1 (TGFbeta1), o sistema renina-angiotensina uteroplacentário (SRA) e os fatores angiogênicos, tais como vascular endothelial growth factor A (VEGF-A) participam do processo de placentação e regulação do fluxo sanguíneo uteroplacentário. Assim, o objetivo do presente estudo foi investigar o efeito da exposição à poluição do ar sobre a morfologia, função e SRA placentários. Métodos: Ratas Wistar fêmeas foram expostas ao ar filtrado (F) ou ao material particulado 2.5um (P) durante 15 dias. Depois o cruzamento, as ratas foram divididas em 4 grupos e novamente expostas a F ou P (FF, FP, PF, PP). No 19º dia de gestação, as porções maternas e fetais das placentas foram coletadas. Estrutura da placenta, TGFbeta1, VEGF-A e seus receptores e os componentes do SRA foram avaliados. Resultados: A exposição ao material particulado diminuiu massa, tamanho e área de superfície placentária, um indicativo da interação materno-fetal. As concentrações placentárias de TGF beta1, VEGF-A e Flk-1 e os componentes do SRA foram alterados e isso pode indicar um prejuízo na invasão do trofoblasto, angiogênese placentária e troca de nutrientes e gases entre mãe e fetos. Discussão: Os resultados indicam que a exposição a partículas compromete a interação materno-fetal e pode refletir sobre a nutrição e crescimento fetal.

Introduction: Exposure to air pollution during pregnancy causes alterations in placental characteristics and may result in intrauterine growth restriction. It was suggested that transforming growth factor beta 1 (TGFbeta1), the uteroplacental renin-angiotensin system (RAS) and angiogenic factors, such as vascular endothelial growth factor A (VEGF-A) participates of the placentation process and regulation of the uteroplacental blood flow. Thus, the aim of the present study was to investigate the effect of exposure to air pollution on the placental morphology, function and placental RAS. Methods: Female Wistar rats were exposed to filtrated air (F) or to concentrated particulate matter 2.5um (P) for 15 days. After mating, rats were divided in 4 groups and again exposed to F or P (FF, FP, PF, PP). At 19th day of pregnancy, maternal and fetal portions of placenta were collected. Placental structure, TGFbeta1, VEGF-A and its receptors and RAS components were evaluated. Results: Exposure to particulate matter decreased placental mass, size and surface area, an indicative of maternal-fetal interaction. Placental TGFbeta1, RAS components and VEGF-A and receptors Flk-1 concentrations were altered and this may indicate a prejudice in the trophoblast invasion, placental angiogenesis and maternal-fetal nutrients and gases exchange. Discussion: These findings indicate that exposure to particulate matter compromises the maternal-fetal interaction and may reflect on fetal nutrition and growth.

[Immunohistochemical study of Dlx1 and Msx1 expression during cephalic development of Dumbo and Wistar rats. Correlation with morphological data]. / Étude immuno-histochimique de l'expression des protéines Dlx1 et Msx1 durant la céphalogenèse des rats Wistar et Dumbo Corrélation avec les données morphologiques.

The Dumbo rat is characterized by a short snout, low ears and relative hypoplasia of maxillar and zygomatic bones. It corresponds to an autosomal recessive genotype. Previous study demonstrated a global deficit of Dlx1 and Msx1 genes expression in comparison to Wistar embryos as considered as control animals. We performed a histological study of cephalic development of Dumbo rats compared to Wistar embryos and an immunohistochemical analysis of Dlx1 and Msx1 protein expression during cephalogenesis. Our data indicate that the pattern of expression of both genes is similar in both strains, but that quantitative differences in gene expression can be the result of delayed organogenesis in Dumbo rat in comparison to Wistar. Some data about gene expressions are discussed at the light of the postulated function of Dlx1 and Msx1 in cephalic development.

Cysteine residues in the large extracellular loop (EC2) are essential for the function of the stress-regulated glycoprotein M6a.

Gpm6a was identified as a stress-responsive gene in the hippocampal formation. This gene is down-regulated in the hippocampus of both socially and physically stressed animals, and this effect can be reversed by antidepressant treatment. Previously we showed that the stress-regulated protein M6a is a key modulator for neurite outgrowth and filopodium/spine formation. In the present work, mutational analysis was used to characterize the action of M6a at the molecular level. We show that four cysteines 162, 174, 192, and 202 within EC2 are functionally crucial sites. The presence of cysteines 162 and 202 is essential for the efficient cell surface expression of the M6a protein. In contrast, cysteines 174 and 192, which form a disulfide bridge as shown by biochemical analysis, are not required for the efficient surface expression of M6a. Their mutation to alanine does not interfere with the localization of M6a to filopodial protrusions in primary hippocampal neurons. The neurons expressing C174A and/or C192A mutants display decreased filopodia number. In non-permeabilized cells, these mutant proteins are not recognized by a function-blocking monoclonal antibody directed to M6a. Moreover, neurons in contact with axons expressing C174A/C192A mutant display significantly lower density of presynaptic clusters over their dendrites. Taken together, this study demonstrates that cysteines in the EC2 domain are critical for the role of M6a in filopodium outgrowth and synaptogenesis.

Chronic stress effects on NPY neuronal population during rat development / Efectos del stress crónico sobre la población neuronal NPY durante el desarrollo de ratas

The aim of this work was to determine the chronical stress effects on the encephalic NPY neurons population during the fetal Central nervous system development. Immunocytochemical techniques were used for this purpose: NPY neurons presented a similar morphology during the gestation days studied but their distribution varied in the anterior, medium and posterior brain. Statistical Highly significant differences in number of NPY positive neurons (p<0.01) among anterior, medium and posterior brain of stressed fetus (SF) were determined depending on the gestation period and the brain area. The NPY neurons were increased in ARC (Arcuate Hypothalamic Nucleus), PH (Posterior Hypothalamic Area) and DM (Dorsomedial Hypothalamic Nucleus) in stressed fetuses (SF) of 17 days, and in ARC of 19 days SF (p< 0.01) were detected in the different brain nucleus. The NPY population increased in PnO (Pontine Reticular Nu, Oral Part) and RITg (Reticulotegmental Nu of the Pons) of 17 days SF, while they were detected in posterior brain at Pyx (Pyramidal Decussation), Rob (Raphe Obscurus Nucleus) and RPA (Raphe Pallidus Nucleus) in SF of 19 days. They also increased in number (p<0.05) in DPGI (Dorsal Paragigantocellular Nu), CGPn (Central Gray of Pons) and PrH (Prepositus Hypoglossal Nucleus) of 17 days SF. Finally, any statistical differences were found among CF and SF in the following nuclei: anterior brain, AH (Anterior Hypothalamic Nucleus), DM (Dorsomedia L Hypothalamic Nucleus) of 17 days; ME (Median Eminence)., VMH (Ventromedial Hypothalamic Nucleus) of 19 days; medium brain in CG (Central Periaqueductal Gray), DR (Dorsal Raphe Nucleus) of 17 days and posterior brain in PnC (Pontine Reticular Nu, Caudal Part), PrH (Prepositus Hypoglossal Nucleus), RMgG (Raphe Magnus Nucleus), IO (Inferior Olive) of 17 days. The increase number of NPY neurons found in the stressed rat fetuses in all periods studied would indicate the participation of the NPY System in...

El propósito del presente estudio fue determinar los efectos del estrés crónico en la población de neuronas NPY encefálicas durante el desarrollo del S.N.C. fetal mediante técnicas inmunocitoquímicas. Se demostró que las neuronas NPY presentan un morfología similar en los días de gestación estudiados, pero su distribución varía en el cerebro anterior, medio y posterior. Se comprobaron diferencias altamente significativas entre el cerebro anterior, medio y posterior (p<0,01) de fetos estresados (FE), variando dicha significación dependiendo del día de la gestación y del área estudiada. En los diferentes núcleos cerebrales del cerebro anterior se detectaron aumentos en ARC (Arcuate Hypothalamic Nucleus), PH (Posterior Hypothalamic Area) de 17 días y DM (Dorsomedia L Hypothalamic Nucleus) y en ARC (Arcuate Hypothalamic Nucleus) de 19días (p<0,01) de F.E. En el cerebro medio se detectaron aumentos en DR (Dorsal Raphe Nucleus) (p<0,01) y PN (Pontine Nucleus) (p<0,05) de 19 F.E. En el cerebro posterior se detectaron aumentos en PnO (Pontine Reticular Nu, Oral Part) y RITg (Reticulotegmental Nu of the Pons) de 17 F. E. y Pyx, (Pyramidal Decussation), Rob (Raphe Obscurus Nucleus) y RPA (Raphe Pallidus Nucleus) de 19 F.E. Asimismo se comprobaron aumentos (p<0,05) en DPGI (Dorsal Paragigantocellular Nu.) de 17 F.E, CGPn (Central Gray of Pons) y PrH (Prepositus Hypoglossal Nucleus), de 19 F.E. Finalmente, no se comprobaron diferencias entre F. C. (fetos controles) y F. E. en los siguientes núcleos del cerebro anterior: AH (Anterior Hypothalamic Nucleus), DM (Dorsomedia L Hypothalamic Nucleus), de 17 días; y EM, (Median Eminence), VMH (Ventromedial Hypothalamic Nucleus) de 19 días. En el cerebro medio CG, (Central Periaqueductal Gray), DR (Dorsal Raphe Nucleus) de 17 días. En el cerebro posterior el PnC, (Pontine Reticular Nu, Caudal Part), PrH (Prepositus Hypoglossal Nucleus), RMgG (Raphe Magnus Nucleus), IO (Inferior Olive) de 17 días del cerebro posterior...

Morphological aspects of the embryonic development of the TMJ in rats (Rattus norvegicus albinus) treated with fluoxetine / Aspectos morfológicos del desarrollo embrionario de la ATM en ratas (Rattus norvegicus albinus) tratadas con fluoxetina

The TMJ has been to the Dental community a key point in the search of knowledge, being it part of the temporomandibular joint complex and of the estomatognathic system which are in charge of the mastication, speech, swallowing, as well as participation in breathing and taste perception. For the majority of the women in serious state of depression, which do not respond psychotherapeutic treatment, pharmacological treatment it's applied. The antidepressants serotonin selective reuptake inhibitors (SSRIs) are the most recommended in these cases. The teratogenic effect of the SSRIs is considered controversial, studies done with women who used these drugs during the pregnancy showed that the respiratory and central nervous systems are the most affected, was also recorded a deficit of body growth and the decrease of the encephalon and skull measures. In the present study, our goal was to assess whether the administration of Fluoxetine during the pregnancy modified the embryology and morphology of the TMJ of rats. For that, 16 Wistar female rats from the Nutrition Department of the UFPE vivarium were selected; 8 for the control group, which received daily 0.9 percent of saline in subcutaneous dose of 10ml/g, with schedules previously established after daily weighing and 8 for the experimental one that were treated with fluoxetine hydrochloride with the dose of 10mg/Kg in a volume 10ml/g of weight, were injected subcutaneously with the same standards established for the control group. It was observed, with this dose that the embryological development of the TMJ, especially of the mandibular condyle, does not present any difference between the degree of maturation of the tissue that forms the TMJ, especially of the condyle between the treated and control groups.

La ATM ha sido para la comunidad odontológica un punto clave en la búsqueda del conocimiento, dado que forma parte del complejo articular temporomandibular y del sistema estomatognático, los cuales se encargan de la masticación, fonación y deglución, así como la participación en la respiración y de la percepción gustativa. Para la mayoría de las mujeres con cuadros graves de depresión, que no responden al tratamiento psicoterapéutico, el tratamiento farmacológico es aplicado. Los antidepresivos del grupo de los Inhibidores Selectivos de Recaptación de Serotonina (ISRSs) son los más comúnmente recomendados en estos casos. El efecto teratogénico de los ISRSs es considerado controversial. Estudios realizados en mujeres que utilizaron estas drogas durante la gestación mostraron que los sistemas respiratorios y nervioso central son los más afectados, también fue constatado un déficit de crecimiento corporal, encefálico y disminución de las medidas craneales. En el presente estudio, nuestro objetivo fue evaluar si la administración de fluoxetina durante la gestación modifica la embriología y la morfología de la ATM de ratas de laboratorio. Para este fin, 16 ratas Wistar del bioterio de nutrición de la UFPE fueron seleccionadas: 8 para el grupo de control, las cuales recibieron diariamente solución fisiológica a 0,9 por ciento en aplicaciones subcutáneas en la dosis de 10ml/g, en horarios previamente establecidos después de pesaje diario y 8 para el experimental, las que fueron tratadas con clorhidrato de fluoxetina en la dosis de 10mg/kg, en un volumen de 10ml/g, inyectados por vía subcutánea en los mismos estándares establecidos para el grupo de control. Se observó, que con esta dosis el desarrollo embriológico de la ATM, especialmente del cóndilo mandibular, no presentó ninguna diferencia entre el grado de maduración de los tejidos que forman la ATM, especialmente del cóndilo, entre los grupos tratado y grupo control.

The effects of valproic acid on sciatic nerve of fetal rats and protective effects of folic acid and vitamin E / Efectos del ácido valproico sobre el nervio ciático de fetos de ratas y efectos de protección del ácido fólico y vitamina E

We aimed to investigate the potential harmful effects of maternal valproic acid (VPA) on fetal sciatic nerve, and the protective effects of vitamin E (Vit E) and folic acid (FA) on fetal rats. Valproic acid (400mg/kg), folic acid (400mg/kg) and vitamin E (250 mg/kg) were administered to rats on each of gestation days 8-10. All fetuses were collected on gestation day 20. With thin sections of biopsies, sciatic nerve of fetuses were stained with uranyl acetat and were examined under transmission electron microscope. The fetuses (n:36) were divided into five groups: control, vpa, vpa+fa, vpa+vit e and vpa+fa+vit e groups. In each group; drug procedure, surgical procedure and histological methods were performed. Later, weights and lengths of fetuses in each group were compared and analyzed by One-Way Anova test. Administration of single doses of valproic acid (400 mg/kg) resulted in weight and length loss between control and vpa group. However, length and weight differences between the other groups were not significant. The histopathological findings of control group was normal. In vpa group, it showed extensive degenerative changes especially in myelin coat. In addition, most prominent finding in this group was condensation of collagen fibers in extensively demyelinated samples, while moderately effected areas were relatively normal. Both vpa+fa and vpa+ vit e groups exhibited similar ultrastructural changes, reflecting minimal to moderate degenerative changes. In vpa+fa+vit e group had almost the normal structure. Administration of single doses of valproic acid (400 mg/kg) resulted in a deteriorative effect on sciatic nerve at ultrastructural level. Administration of FA and Vit E had a protective effect to prevent the degenerative changes to a certain degree. Combination of FA and Vit E together following VPA administration had a more potent protective effect. The objective of the present study is to analyze histopathologic changes which ...

El objetivo fue investigar los posibles efectos perjudiciales del ácido valproico (AVP) materno sobre el nervio ciático en fetos y los efectos protectores de la vitamina E (Vit E) y ácido fólico (AF) en fetos de ratas. Se administraron a ratas ácido valproico (400mg/kg), ácido fólico (400mg/kg) y vitamina E (250 mg/kg) en cada uno de los días de gestación 8-10. Todos los fetos fueron recogidos a los 20 días de gestación. Finas secciones de biopsias obtenidas de los nervios ciáticos de fetos fueron teñidos con acetato de uranilo y examinados bajo microscopio electrónico de transmisión. Los fetos (n: 36) fueron divididos en cinco grupos: control, avp, avp+af, avp+vit e y avp+fa+vit e. En cada grupo, se realizaron los procedimientos farmacológicos, quirúrgicos y los métodos histológicos. Los pesos y longitudes de los fetos de cada grupo fueron comparados y analizados usando la prueba One-Way Anova. La administración de dosis únicas de ácido valproico (400 mg / kg) resultó en la pérdida del peso la longitud entre el control y el grupo apv. Sin embargo, las diferencias en la longitud y el peso entre los otros grupos no fueron significativas. Los hallazgos histopatológicos del grupo control fueron normales. En el grupo avp, se mostró especialmente cambios degenerativos en la mielina que envuelve al nervio periféricamente. Además, predominatemente se encontró en las muestras de este grupo fibras colágenas condensadas y zonas ampliamente desmielinizadas, mientras que las zonas moderadamente afectadas eran relativamente normales. Ambos grupos avp+fa y avp+vit e exhibieron cambios ultraestructurales similares, lo que supone un mínimo o moderado cambio degenerativo. El grupo avp+fa+vit e tuvo casi una estructura normal. La administración de dosis únicas de ácido valproico (400 mg / kg) produjo un efecto sobre el deterioro del nervio ciático a nivel ultraestructural. La administración de la AF y vitamina E tienen un efecto protector, en cierta medida, ...

Effect of anti-basic fibroblast growth factor (anti-bFGF) on in vitro embryonic development in rat.

In this study, we aimed at the in vitro effects of anti-fibroblast growth factor-2 (anti-FGF-2 or anti-bFGF) on embryo culture in rats. In vitro effects of anti-bFGF on total embryonic development were investigated in 40 rat embryos (which were divided into four groups) (obtained from five pregnant females) at 9.5 days of gestation that were cultured in whole rat serum (WRS), and in WRS+ 2.5, 5, and 10 microg/ml anti-bFGF. After 48 h of culturing, the embryos from each group were harvested to be analysed morphologically according to a morphological scoring system and biochemically to obtain the embryo protein content. The morphological score, embryo protein content, somite number and crown-rump length of embryos indicated that embryos cultured in WRS+ anti-bFGF had significant embryonic retardation. Mean morphological scores for the embryos grown in WRS, in the presence of 2.5, 5 and 10 microg anti-FGF-2 were 61.4 +/- 1.64, 46.3 +/- 8.42, 27 +/- 2.58 and 13.6 +/- 0.96 respectively. These results suggest that bFGF is very important for normal embryonic development and rat anti-bFGF neutralizes bFGF effect.

Investigation of direct toxic and teratogenic effects of anticoagulants on rat embryonic development using in vitro culture method and genotoxicity assay.

Heparin and low molecular weight heparins (LMWHs) are used to reduce the incidence of venous thromboembolism in pregnancy. Although, these agents have been shown to be safe when used during pregnancy, the studies about direct toxic and teratogenic effects of these drugs on embryonic development are limited. In this study, the effects of heparin and LMWHs on rat embryonic development were investigated by using in vitro embryo culture and micronucleus (MN) assay methods. Rat embryos were cultured in vitro in the presence of different concentrations of heparin (5-40 IU/ml), dalteparin (2.5-20 IU/ml), enoxaparin (25-100 microg/ml) and nadroparin (1-4 IU/ml). Effects of anticoagulants on embryonic developmental parameters were compared and embryos were evaluated for the presence of any malformations. After culturing the embryos, classic MN assay was performed. Anticoagulants significantly decreased all growth and developmental parameters dose-dependently. Dalteparin and enoxaparin were found to cause more developmental toxicity than heparin and nadroparin. Along with haematoma in general, heparin and nadroparin caused maxillary deformity, situs inversus and oedema most frequently, while neural tube defects were observed with dalteparin and enoxaparin. All agents also significantly induced MN formation in rat embryonic blood cells. These results indicate the possible genotoxic effects of anticoagulant agents on the developing rat embryo when applied directly.

Verificação do efeito do Lapachol sobre o sistema morfogenético em ratas prenhes / Assessment of lapachol's on the morphogenetic system of pregnant rats

Estudos prévios com o lapachol, administrados durante o período de organogênese, indicaram que o fitofármaco é embriotóxico. A exposição de ratas prenhes a xenobióticos por longo período pode causar toxicidade materna responsável por alterações morfológicas do concepto. No presente trabalho avalia-se o efeito do lapachol, administrado no nono dia de prenhez, sobre o organismo materno e o sistema morfogenético do concepto. Ratas Wistar foram distribuídas em quatro grupos: controle (1 ml de água destilada), veículo (1 ml de solção hidroalcoólica), lapachol 100 (L - 100) e 200 (L - 200) mg/kg peso corporal. Tratamento por via gástrica em dose única. Mediu-se peso corporal e consumo de ração. No 21º dia os animais foram sacrificados por exsanguinação total sob anestesia. No soro foram dosadas TGP e uréia. Procedeu-se á autópsia e á remoção de fígado, rins (pesados e fixados para análise anatomopatológica) e trato reprodutor materno. O vários foram pesados e os corpos lúteos contados. Nos cornos uterinos contaram-se implantes, reabsorções, e fetos mortos. Fetos e placentas foram pesados. Ratas tratadas com L - 200 tiveram peso de fígado e de ovários menor que os demais grupos; aumento da concentração de TGP e de uréia e 83% de mortes de conceptos. Ratas tratadas com L - 100 tiveram 32% de mortes de conceptos. os resultados indicam possibilidade de toxicidade materna com a dose maior de lapachol. Pode-se concluir que o lapachol ou seus produtos biotransformados, agem diretamente sobre o sistema morfogenético do embrião e doses elevadas podem ter seu efeito potencializado por toxicidade materna leve.

Palatal abnormalities in the developing rat induced by retinoic acid

Palatogenesis is a complex developmental processthat requires two main events: elevation andthen fusion of the palatal shelves. These processesare disrupted by teratogens such as retinoicacid (RA) and genetic defects, resulting in variousmalformations (including cleft palate). Usinghistological and immunohistochemical techniques,the effects of different isomers of RA, administeredin various concentrations to pregnantrats on different gestational days (GD), were assessedfrom observations of the state of palataldevelopment on GD 18 in foetuses without exencephaly.Varying degrees of clefting of thepalate were observed, from failure of elevationof the palatal shelves to failure of fusion in themidline. This study shows that all-trans-RA isthe most teratogenic RA isomer in terms of ratpalatal abnormalities. It also supports previousfindings that the timing of administration of alltrans-RA is more critical than the concentration,with treatment between GD 10 and 10.5 havingthe most severe effects. Previous histologicalstudies also suggested that RA is associatedwith the appearance of ectopic cartilages withinthe developing palate of foetuses showing exencephaly.In this investigation, immunohistochemicallabelling of the foetal material with antibodiesthat recognise epitopes present in linkproteins 1, 2, and 3 (8A4), chondroitin-4-sulphatestubs (2B6), and G1 and chondroitin sulphateattachments (7D1) present in aggrecan (associatedwith hyaluronan in cartilage) showedno signs of ectopic cartilage formation within the palate at GD18. Internal controls of the cartilagesof the nasal septum, vomeronasal cartilage, andMerkel’s cartilage labelled intensely and appearedmorphologically normal (AU)

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Functional interaction between neuropeptide Y receptors and modulation of calcium channels in the rat hippocampus.

We investigated the functional interaction between neuropeptide Y (NPY) receptors using nerve terminals and cultured rat hippocampal neurons, and we evaluated the involvement of voltage-gated Ca(2+) channels (VGCCs) in NPY receptors-induced inhibition of Ca(2+) influx and glutamate release. The KCl-evoked release of glutamate from hippocampal synaptosomes was inhibited by 1 microM NPY and this effect was insensitive to either BIBP3226 (Y1 receptor antagonist) or L-152,804 (Y5 receptor antagonist), but was sensitive to BIIE0246 (Y2 receptor antagonist). We could also pharmacologically dissect the NPY receptors activity by using Y1, Y2 and Y5 receptor agonists ([Leu(31),Pro(34)]NPY, NPY13-36, NPY (19-23)-(Gly(1),Ser(3),Gln(4),Thr(6),Ala(31),Aib(32),Gln(34))-pancreatic polypeptide (PP), respectively), and in all the cases we observed that these agonists could inhibited the KCl-induced release of glutamate. However, the selective and specific co-activation of both Y1 and Y2 or Y2 and Y5 receptors resulted in non-additive inhibition, and this effect was prevented in the presence of the Y2 antagonist, but was insensitive to the Y1 or Y5 receptor antagonist. Moreover, as we previously showed for Y1 receptors, we also observed that the activation of Y5 receptors inhibited the glutamate release in the dentate gyrus and CA3 subregion, without significant effect in the CA1 subregion of the hippocampus. The same qualitative results were obtained when we investigated the role of NPY Y1 and Y2 receptors in modulating the changes in [Ca(2+)](i) due to KCl depolarisation in cultured hippocampal neurons. The inhibitory effect of nitrendipine (L-type VGCC blocker) or omega-conotoxin GVIA (omega-CgTx; N-type VGCC blocker) was not potentiated by the simultaneous activation of Y1 or Y2 receptors. Moreover, the exocytotic release of glutamate was inhibited by omega-agatoxin IVA (omega-Aga; P-/Q-type VGCC blocker), and this VGCC blocker did not potentiate Y1, Y2 or Y5 receptor-mediated inhibition of glutamate release. Also, the effect of ionomycin in inducing the exocytotic release of glutamate from hippocampal synaptosomes was insensitive to the activation of NPY receptors. In the present paper, we identified a role for NPY Y1, Y2 and Y5 receptors in modulating the exocytotic release of glutamate and the [Ca(2+)](i) changes in the rat hippocampus. In conditions of co-activation, there appears to exist a physiological cross-talk between Y1 and Y2 and also between Y2 and Y5 receptors, in which Y2 receptors play a predominant role. Moreover, we also show that Y1 and Y2 receptors exert their inhibitory action by directly modulating L-, N-, and P-/Q-type VGCCs, whereas the inhibition of glutamate release mediated by the Y5 receptors seems to involve P-/Q-type VGCCs.