Talin1 is highly expressed in the fallopian tube and chorionic villi to promote trophoblast invasion in tubal pregnancy / 南方医科大学学报

OBJECTIVE@#To investigate the expression of Talin1 in the fallopian tube and chorionic villi in patients with tubal pregnancy and its role in regulating invasion and migration of trophoblasts.@*METHODS@#Immunohistochemistry and Western blotting were used to detect the localization and expression level of Talin1 in the fallopian tube and chorionic villi in patients with tubal pregnancy and in women with normal pregnancy. In the cell experiment, HTR-8/SVneo cells was transfected with Talin1 siRNA and the changes in cell invasion and migration were assessed using scratch assay and Transwell assay. The expressions of MMP-2, MMP-9, N-cadherin and Snail in the transfected cells were detected by qRT-PCR and Western blotting.@*RESULTS@#Positive expression of Talin1 was detected in both normal fallopian tube tissues and tissues from women tubal pregnancy, and its expression was localized mainly in the cytoplasm of cilia cells. The expression level of Talin1 was significantly higher in both the fallopian tube and chorionic villi in women with tubal pregnancy than in normal fallopian tube and chorionic villi samples (P < 0.01). In HTR-8/SVneo cells, transfection with Talin1 siRNA significantly inhibited cell invasion (P < 0.01) and migration (P < 0.05), down-regulated the expression of N-cadherin, MMP-2 and Snail (P < 0.05), and up-regulated the expression of MMP-9 in the cells (P < 0.05).@*CONCLUSION@#The expression of Talin1 in the fallopian tube and chorionic villi is significantly increased in women with tubal pregnancy, suggesting the association of Talin1-regulated trophoblast cell invasion with the occurrence of tubal pregnancy.

Expresión de calreticulina en placenta canina / Calreticulin expression in canine placenta

La placenta es un anexo embrionario de los mamíferos que tiene por función principal el intercambio de nutrientes y gases y proteger al concepto de un potencial daño inmune provocado por diferencias alogénicas en los Complejos Principales de Histocompatibilidad paternos. Se han descrito diversas proteínas asociadas a su función, siendo Calreticulina una de ellas. Si bien existen estudios de la presencia de Calreticulina en placenta humana, no existen reportes de esta proteína en la placenta canina. Se obtuvieron muestras de placenta canina de las que se extrajo el contenido proteico total y se determinó la presencia de Calreticulina por western blot e inmunohistoquímica. Los resultados mostraron presencia de Calreticulina en placenta canina con un peso molecular aparente de 60 kDa, concordante con lo descrito para la molécula por otros autores. El análisis inmunohistoquímico mostró que Calreticulina canina está presente principalmente en el trofoblasto de las vellosidades, no existiendo diferencias en cuanto a su localización al compararla con placenta humana, pese a sus diferencias morfológicas e histológicas. Esta información permitirá establecer un protocolo estandarizado de extracción de Calreticulina desde placenta, así como orientar acerca de los posibles roles de esta molécula en la placenta.

The placenta is an embryonic organ present in mammals, whose main functions are the exchange of nutrients and gases and to protect the fetus from potential immune damage mediated by paternal and maternal allogeneic differences in the Major Histocompatibility Complex. Several proteins associated with its function have been described, being Calreticulin one of them. Although there are studies on the presence of Calreticulin in human placenta, there are no reports of this protein in canine placenta. Samples from canine placenta were obtained, proteins extracted and Calreticulin was subsequently detected by western blot and immunohistochemistry. The results showed the presence of Calreticulin in canine placenta with an apparent molecular weight of 60 kDa, in agreement with the results from other authors. The immunohistochemical analysis showed that canine Calreticulin is present mainly in the trophoblast of the villi, and there is no difference in its localization when compared with a blood-filled placenta such as human one, despite its morphological and histological differences. We also propose a standardized protocol for the extraction of Calreticulin from placenta, given its abundant expression in this organ. Future studies are aimed at elucidating possible roles of this protein in placenta.

Fisiopatologia da pré-eclâmpsia / Pathophysiology of preeclampsia

A pré-eclâmpsia é uma complicação da gravidez que ocorre no terceiro trimestre de gestação e é específica da gravidez humana. Ela é secundária a uma disfunção placentária, a qual é responsável pela liberação na circulação materna de diversas substâncias responsáveis por ativação e até lesão do endotélio materno, tendo como consequências hipertensão arterial, nefropatia glomerular e aumento da permeabilidade vascular. Trabalhos recentes têm permitido melhor compreensão da fisiopatologia desta patologia, com a esperança de desenvolver novos instrumentos de detecção e possíveis perspectivas terapêuticas. Este artigo ilustra os mecanismos fisiopatológicos da revascularização uterina defeituosa, da disfunção placentária e da disfunção endotelial que ocorrem no decurso da pré-eclâmpsia.

Estudio cariométrico de placentas de ratones con infección aguda por diferentes cepas de Trypanosoma cruzi / Karyometric study of placentas from mice with acute infection by different strains of Trypanosoma cruzi

El objetivo de este trabajo fue evaluar cariometricamente las alteraciones causadas por diferentes cepas de T. cruzi en la placenta del ratón. Ratones hembras de 60 días, grávidas, fueron inoculadas, intraperitonealmente, con 2 x 10(5) tripomastigotes sanguíneos de las cepas colombiana, Y, Solivia o RC del T. cruzi. Fueron observadas claras diferencias en las alteraciones cariométricas de las células trofoblásticas gigantes y de las células trofoblásticas de la zona esponjosa. Los resultados demostraron que las cepas colombiana y RC causan alteraciones tanto en las células trofoblásticas gigantes como en las células del trofoblasto esponjoso, mientras que las cepas Y y Bolivia provocan alteraciones solamente en las células trofoblásticas gigantes. Es posible concluir que cada cepa posee características propias y que, a pesar del tipo similar de transmisión, presenta matices diferenciales en el proceso de la patogénesis placentaria.

The objective of this work was to evaluate karyometrically the alterations caused by different strains of Trypanosoma cruzi in the mouse placenta. Pregnant mice, 60-day old, were intraperitoneally inoculated with 2 x 10(5) bloodstream trypomastigotes of the Colombian, Y, Bolivia or RC strain of T cruzi. There were observed clear differences in the karyometric alterations of the trophoblast giant cells and in the spongiotrophoblast cells. The results demonstrate that the Colombian and RC strains cause alterations both in the trophoblast giant cells and in the spongiotrophoblast cells, whereas the Y and Bolivia strains provoke alterations only in the trophoblast giant cells. It is possible concluding that each strain has its own characteristics and that, in spite of the similar type of transmission, it show differential nuances in the placental pathogenic process.

Apical Maxi-chloride channel from human placenta: 12 years after the first electrophysiological recordings

The Maxi-chloride channel was the first ion channel described by electrophysiological methods in placenta. Because it is difficult to access a complex epithelium such as the placenta for electrophysiological procedures, the studies of ion channels from placental membranes have been performed only very recently. It was only in 1993 that a direct demonstration of a high-conductance chloride channel in apical membranes of intact trophoblastic epithelium was mentioned, and two years later, the description of this channel was reported from purified placental apical membranes reconstituted into artificial lipid membranes suitable for patch-clamp recordings. This brief review comments on the work done with regard to the electrophysiological characterization and regulation of the large-conductance or "Maxi" chloride channel and its contribution to the development of a cellular model for syncytiotrophoblast ion transport.

Serum beta human chorionic gonadotropin levels and preeclampsia

Preeclampsia is one of the most common complications of pregnancy. It is a trophoblastic disorder. Since human chorionic gonadotropin [hCG] is secreted by trophoblast, the purpose of the study was to determine the levels of beta [hCG] in preeclampsia and normal cases. This study was conducted on 40 term pregnant women with preeclampsia [case group] and 40 normal pregnant cases [control group] in the Department of Obstetrics and Gynecology, Medical University, Babol, Iran, from 2004 to 2005. Serum beta-hCG in all cases were measured by radioimmunoassay. The mean beta-hCG level in preeclampsia [mild and severe] was compared with normal cases. The maternal serum, beta-hCG level in patients with preeclampsia [39840 +/- 24630 IU/L] was higher than in the control group [27460 +/- 25862 IU/L] [p=0.031]. In mild preeclampsia [34 cases], the mean serum beta-hCG level was 31991 +/- 16758 IU/L while in severe preeclampsia [6 cases] 84312 +/- 9257 IU/L [p<0.0001]. The mean serum beta-hCG level in mild and severe preeclampsia was higher compared to the normal cases. The results of this study show that the levels of beta-hCG in severe form of preeclampsia is higher compared to the milder ones

Comportamiento clínico y seguimiento de la enfermedad del trofoblasto gestacional abril-1996-dicembre-1999 en el Hospital Bertha Calderón

El presente estudio es de tipo descriptivo decorte longitudinal realizado en el período de abril de 1996 a diciembre de 1999. Se estudiaron 118 pacientes que se ingresaron al servicio de complicaciones I y Oncología: fueron ingresadas por presentar cualquier entidad clínica de la Enfermedad Trofoblastica gestacional; a estas pacientes se les dió seguimiento durante el tiempo que duró el estudio en el área de consulta externa del Hospital Berta Calderón. Se analizó cada uno de los casos tomando como parámetros, el abordaje clínico y de laboratorio. La enfermedad de Trofoblasto afecta especialmente a las mujeres jóvenes en edad reproductiva, menor de 25 años no existiendo diferencia respecto a la aparición de la enfermedad con la gestación. En su mayoría consultaron por sangrado transvaginal asociado con dolor bajo vientre o hiperemesis gravídica. Acudieron con el útero de mayor tamaño de acuerdo a su edad gestacional. El 77 porciento de las pacientes fueron diagnósticadas por ultrasonido pélvico. La cuantificación de la Hormona Gonodotropina Coriónica Humana en el 64 porciento de los casos no fue realizada; siendo la causa principal el factor económico. El 97 porciento de los casos se evacuó por legrado por aspiración eléctrico, confirmándose histopatológicamente como Mola Hidatidiforme completa en el 86 porciento de los casos 2.5 como coriocarcino, 1.6 porciento como Mola Parcial y Mola Invasiva respectivamente. En cuanto a la estadificación de riesgo se encontró 7 casos de bajo riesgo, y 3 de alto riesgo. Se trato a las pacientes de bajo riesgo con mono quimioterapia (Methodrexate o Actinomicina D). Los altos riesgos 3 caso; 2 de ellos se trataron con poli quimoterapia (MAC o EMACO), que además se asoció con histerectomía abdominal, una por sangrado profuso y otra por que el foco tumoral In útero. El tercero caso se le indico la poli quimoterapia, pero no se le aplicó por malas condiciones de la pacientes y falleció antes de aplicarse el tratamiento. Se considera que el tratamiento indicado fue satisfactorio en las pacientes que acudieron a su consulta y que los niveles de Hormona Gondotropina Coriónica Humana llegaron a niveles normales. El 54 porciento de las pacientes estudiadas acudieron por lo menos a un control de seguimiento en la consulta externa; el 6 porciento de éstos acudieron a sus 8 controles...

Modulation of bilayer fluidity by lipid peroxidation of human placental syncytiotrophoblast membranes during embryogenesis.

Placental syncytiotrophoblast while regulating the passage of nutrients from maternal blood to the fetal circulation exposes itself to the risk of oxidative attack by the oxygen free radicals. Extent of lipid peroxidation (LPO) investigated in placental brush border membrane (BBM) and basal membrane (BM) revealed a decreasing trend with gestational progress. Steady-state fluorescence anisotropy measurement of 1,6-diphenyl-1,3,5-hexatriene (DPH)-labeled malondialdehyde treated placental membrane vesicles suggested modulation of BBM and BM fluidity by lipid peroxidation in all gestational ages. alpha-tocopherol content in both the placental membranes which increased as gestation progressed has been proposed to play a significant role in decreasing LPO of placental membranes during intrauterine development.

Biologically active steroid and thyroid hormones stimulate secretion of sex hormone-bending globulin by human term placenta in culture

In this study, the influence of steroid and thyroid hormones and epidermal growth factor on the productin of SHBG by placenta tissue explants was investigated. Explants of trophoblastic tissue obtained from normal term placentas were cultured for 48 h in serum free culture medium, and then for an additional 24 h period in the presence or absence of various concentrations of either estradiol (0.25 - 5 nM), testoterone (0.5 - 500 nM), triiodothyronine (0.01 - 100 nM) or EGF (2-40 µM), respectively. Human SHBG concentration in culture media was estimated on each day by specific two-site time-resolved fluoroimmunometric assay and the results expressed as pmol/mg tissue protein. Binding characteristics and molecular structure of secreted SHBG were determined by [3H]5a-DHT binding assays and Western blot analysis, espectively. Estradiol and triidothyronine but not testoterone increased significantly (p<0.05 vs. control) the secretion of SHBG into the culture media. Addition of EGF did not significantly change the production of SHBG at the various concentrations studied. [3H]5a-DHT binding assays and Western blot analysis of placental SHBG resulted in identical binding affinities (Kd 2.0 ñ 0.16 x 10-9M= and molecular structure to those obtained in serum from normal pregnant women. These findings support and extend previous observations by our laboratory indicating that SHBG gene is expressed in the placenta and provide further evidence on the hormonal regulatory characteristics of this steroid-binding protein in cultured placenta

Reactive oxygen species and the phagocytosis process of hemochorial trophoblast

The establishment of the hemochorial placenta requires the trophoblast to invade the endometrium and its vascular network to reach the abundant supply of maternal blood during embryo implantation. To facilitate the invasion steps and/or to provide the nutritional source for embryo development before the completion of the placenta, the trophoblast exhibits a temporally limited and very intense phagocytic activity. Despite the importance of this critical event during pregnancy, full understanding of the molecular mechanisms involved is still to be achieved. Recent findings of our laboratory show that mouse trophoblast cells are able to produce and release reactive oxygen species (ROS) to the surrounding medium, in a manner similar to activated inflammatory phagocytic cells. The aim of the present communications, is to discuss the results obtained by our group regarding the involvement of ROS in the trophoblast phagocytosis, with special attention to the underlying physiological mechanisms related to the embryo implantation.

Effects of dipyrone on the placenta of albino rats: morphological and biochemical aspects

The effect of dipyrone (N-2,3-dimethyl-5-oxo-1-phenyl-3-pyrazolin-4-yl)-methyllamino methanesulfonate sodium monohydrate) on the placenta of 2 BAW albino rats was studied through Karyometry of trophoblastic giant cells and DNA, RNA and total protein determinations. The animals received a single daily dose of 50 mg/Kg body weight during different periods of pregnancy: from the 9th to the 12th, 11th to the 14th, 13th to the 16th, 15th to the 18th and 17th to the 20 thday. Control animals received a single daily dose of 0.5ml distilled water at the same time. Karyometric results showed a statistically significant increase in nuclear volumes of placental cells of rats receiving dipyrone during the first three periods, when compared to control groups. In the two groups that received the drug nearer to term there was no significant difference. Regarding DNA, RNA and total protein determinations, there was a statistically significant difference, for all of them, in the rats that received the drug from the 9th to the 12 th day of pregnancy when compared to the control group. There was no significant difference in the groups that received the drug after that period. The results show that dipyrone had a blocking effect on cell division and that this effect happens mainly in the initial period of placental development