Fetal-Maternal Interactions in the Synepitheliochorial Placenta Using the eGFP Cloned Cattle Model

Background: To investigate mechanisms of fetal-maternal cell interactions in the bovine placenta, we developed a model of transgenic enhanced Green Fluorescent Protein (t-eGFP) expressing bovine embryos produced by nuclear transfer (NT) to assess the distribution of fetal-derived products in the bovine placenta. In addition, we searched for male specific DNA in the blood of females carrying in vitro produced male embryos. Our hypothesis is that the bovine placenta is more permeable to fetal-derived products than described elsewhere. Methodology/Principal Findings: Samples of placentomes, chorion, endometrium, maternal peripheral blood leukocytes and blood plasma were collected during early gestation and processed for nested-PCR for eGFP and testis-specific Y-encoded protein (TSPY), western blotting and immunohistochemistry for eGFP detection, as well as transmission electron microscopy to verify the level of interaction between maternal and fetal cells. TSPY and eGFP DNA were present in the blood of cows carrying male pregnancies at day 60 of pregnancy. Protein and mRNA of eGFP were observed in the trophoblast and uterine tissues. In the placentomes, the protein expression was weak in the syncytial regions, but intense in neighboring cells on both sides of the fetal-maternal interface. Ultrastructurally, our samples from t-eGFP expressing NT pregnancies showed to be normal, such as the presence of interdigitating structures between fetal and maternal cells. In addition, channels-like structures were present in the trophoblast cells. Conclusions/Significance: Data suggested that there is a delivery of fetal contents to the maternal system on both systemic and local levels that involved nuclear acids and proteins. It not clear the mechanisms involved in the transfer of fetal-derived molecules to the maternal system. This delivery may occur through nonclassical protein secretion; throughout transtrophoblastic-like channels and/or by apoptotic processes previously described. In conclusion, the bovine synepitheliochorial placenta displays an intimate fetal-maternal interaction, similar to other placental types for instance human and mouse. © 2013 Pereira et al.

Cytotoxicity of adhesive systems of different hydrophilicities on cultured odontoblast-like cells

This study evaluated the cytotoxicity of experimental adhesive systems (EASs) on odontoblast-like cells. Paper discs (n=132) were impregnated with 10 μL of each EAS-R1, R2, R3, R4, and R5 (in an ascending order of hydrophilicity), followed by photoactivation. R1 and R2 are nonsolvated hydrophobic blends, R3 represents a simplified etch-and-rinse adhesive system, and R4 and R5 represent simplified self-etch adhesive systems. Discs were immersed in Dulbecco's modified Eagle's medium for 24 h to obtain eluates applied on MDPC-23 cell cultures. No material was applied on discs used as control (R0). Cell viability [3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay], total protein (TP) production, alkaline phosphatase (ALP) activity, type of cell death, and degree of monomer conversion Fourier transform infrared (%DC-FTIR) were evaluated. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (α=0.05). Considering R0 (control) as having 100% of cell viability, R1, R2, R3, R4, and R5 reduced the metabolic activity of cells by 36.4, 3.1, 0.2, 21.5, and 65.7%, respectively, but only R1 and R5 differed from R0. Comparing with R0, lower TP production was observed for R1, R4, and R5, while ALP activity decreased for R1 and R5. Necrotic cell death was predominant for all EASs, but only R1, R4, and R5 differed from R0. Only R5 presented a different apoptotic cell death ratio from R0. R1 presented the lowest %DC (ca. 37%), whereas R4 and R5 presented the highest (ca. 56%). In conclusion, R2 and R3 were not toxic to the MDPC-23 cells, suggesting that the degree of hydrophilicity or %DC of the EASs alone were not responsible for their cytopathic effects. © 2013 Wiley Periodicals, Inc.

Morphological and cytochemical changes in synganglion of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) female ticks from exposure of andiroba oil (Carapa guianensis)

Actually, the most used method to control ticks is synthetic acaricides with neurotoxic action. However, the use of these methods presents inconveniences, such as the contamination of the environment and risks to the host's health due to the residual effects. Thus, several studies have been developed aiming to find alternative ways to control these ectoparasites, such as the use of natural compounds with active ingredients, which act controlling some species of plagues in addition to presenting medicinal properties that are beneficial to humans. The present study aimed to analyze the action of andiroba oil (Carapa guianensis) on the synganglion of Rhipicephalus sanguineus semiengorged females through morphological and cytochemical techniques aiming to verify if this natural product have neurotoxic action as the numerous synthetic acaricides. The results showed that andiroba oil interferes in the synganglion through structural and enzymatic changes, which lead the nervous tissue to apoptotic death involving autophagy. Among these changes was observed the emergence of large empty spaces between the perineurium and the cortical region, vacuolated cortex cells and with cell swelling, neural cells with picnotic nuclei or in initial stage of chromatin margination and neuropile with high structural disorganization. Considering these data, it can be concluded that andiroba seed oil can be used as an alternative method in the control of R. sanguineus ticks due to its neurotoxic action. © 2013 Wiley Periodicals, Inc.

Dynamics of bone healing after osteotomy with piezosurgery or conventional drilling - histomorphometrical, immunohistochemical, and molecular analysis

Background: Piezosurgery is an osteotomy system used in medical and dental surgery. Many studies have proven clinical advantages of piezosurgery in terms of quality of cut, maneuverability, ease of use, and safety. However, few investigations have tested its superiority over the traditional osteotomy systems in terms of dynamics of bone healing. Therefore, the aim of this study was to evaluate the dynamics of bone healing after osteotomies with piezosurgery and to compare them with those associated to traditional bone drilling.Methods: One hundred and ten rats were divided into two groups with 55 animals each. The animals were anesthetized and the tibiae were surgically exposed to create defects 2 mm in diameter by using piezosurgery (Piezo group) and conventional drilling (Drill group). Animals were sacrificed at 3, 7, 14, 30 and 60 days post-surgery. Bone samples were collected and processed for histological, histomorphometrical, immunohistochemical, and molecular analysis. The histological analysis was performed at all time points (n = 8) whereas the histomorphometrical analysis was performed at 7, 14, 30 and 60 days post-surgery (n = 8). The immunolabeling was performed to detect Vascular Endothelial Growth Factor (VEGF), Caspase-3 (CAS-3), Osteoprotegerin (OPG), Receptor Activator of Nuclear Factor kappa-B Ligand (RANKL), and Osteocalcin (OC) at 3, 7, and 14 days (n = 3). For the molecular analysis, animals were sacrificed at 3, 7 and 14 days, total RNA was collected, and quantification of the expression of 21 genes related to BMP signaling, Wnt signaling, inflammation, osteogenenic and apoptotic pathways was performed by qRT-PCR (n = 5).Results: Histologically and histomorphometrically, bone healing was similar in both groups with the exception of a slightly higher amount of newly formed bone observed at 30 days after piezosurgery (p < 0.05). Immunohistochemical and qRT-PCR analyses didn't detect significant differences in expression of all the proteins and most of the genes tested.Conclusions: Based on the results of our study we conclude that in a rat tibial bone defect model the bone healing dynamics after piezosurgery are comparable to those observed with conventional drilling. © 2013 Esteves et al.; licensee BioMed Central Ltd.

Bovine herpesvirus type 5 infection regulates Bax/BCL-2 ratio

Bovine herpesvirus 5 (BoHV-5) is an α-herpesvirus that causes neurological disease in young cattle and is also occasionally involved in reproductive disorders. Although there have been many studies of the apoptotic pathways induced by viruses belonging to the family Herpesviridae, there is little information about the intrinsic programmed cell death pathway in host-BoHV-5 interactions. We found that BoHV-5 is able to replicate in both mesenchymal and epithelial cell lines, provoking cytopathology that is characterized by cellular swelling and cell fusion. Viral antigens were detected in infected cells by immunofluorescence assay at 48 to 96 h post-infection (p.i.). At 48 to 72 h p.i., anti-apoptotic BCL-2 antigens were found at higher levels than Bax antigens; the latter is considered a pro-apoptotic protein. Infected cells had increased BCL-2 phenotype cells from 48 to 96 h p.i., based on flow cytometric analysis. At 48 to 96 h p.i., Bax mRNA was not expressed in any of the infected cell monolayers. In contrast, BCL-2 mRNA was found at high levels at all p.i. in both types of cells. BoHV-5 replication apparently modulates BCL-2 expression and gene transcription, enhancing production of virus progeny. © FUNPEC-RP.

Ultrastructural characteristics of the spermatogenesis during the four phases of the annual reproductive cycle of the black myotis bat, Myotis nigricans (Chiroptera: Vespertilionidae)

Myotis nigricans is an endemic species of vespertilionid bat, from the Neotropical region, that resembles temperate zone bats in their reproductive cycle; presenting an annual reproductive cycle with two periods of testicular regression, which are not linked to the apoptotic process and seems to be not directly linked to any seasonal abiotic variation. Thus, this study aimed to ultrastructurally evaluate their reproductive cycle. The process of testicular regression could be divided into four periods: active; regressing; regressed and recrudescence; with all presenting distinct characteristics. The active period was similar to that of other bats, presenting the complete occurrence of spermatogenesis, with three main types of spermatogonia (Ad, Ap, and B) and 12 steps in spermatid differentiation; however, it differed in having the outer dense fibers 1, 5, 6, and 9 larger than the others. These three types of spermatogonia undergo considerable morphologic changes from regressing to the regressed period, and in the recrudescence, they return to the basic morphology, which reactivates spermatogenesis. In conclusion, our study described the process of spermatogenesis, the ultrastructure of the spermatozoa and the distinct morphologic variations in the ultrastructure of the testicular cells of M. nigricans during the four different periods of its annual reproductive cycle. Microsc. Res. Tech., 76:1035-1049, 2013. © 2013 Wiley Periodicals, Inc.

Avaliação da atividade tripanocida do ácido gálico e seus ésteres contra formas epimastigotas de Trypanosoma cruzi: Rogério Andréo. -

Pós-graduação em Biotecnologia - IQ

CD95 (FAS) e CD178 (FASL) induzem apoptose em células CD4+ E CD8+ do sangue periférico e esplênicas em cães naturalmente infectados por Leishmania spp: Kathlenn Liezbeth Oliveira Silva. -

Pós-graduação em Ciência Animal - FMVA

Intervenção dietética com suco de laranja sobre o estado nutricional e oxidativo em pacientes com hepatite C crônica

Pós-graduação em Alimentos e Nutrição - FCFAR

Atividade citotóxica, pró-apoptótica, genotóxica e mutagênica de nitensidina B em células tumorais cervicais humanas

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Avaliação da atividade apoptótica de substância pura isolada de Cryptocarya mandioccanna em células de carcinoma cervical imortalizadas pelo papilomavirus humano (HPV)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Papilomavírus humano e a carcinogênse de mucosa oral: avaliação imunohistoquímica das protínas p27, mdm2 E catepsina B

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação da prliferação e apoptose com a utilização de arranjos em matriz de amostra tecidual (Tissue Microarray - TMA) em mastocitomas cutâneos caninos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeito do soro urêmico de cães com insuficiência renal sobre o metabolismo oxidativo e apoptose dos polimorfonucleares

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo da ação de nanopartículas magnéticas de ferro no câncer oral

Pós-graduação em Genética - IBILCE