Association of early cleavage, morula compaction and blastocysts ploidy of IVF embryos cultured in a time-lapse system and biopsied for genetic test for aneuploidy.

IVF embryos have historically been evaluated by morphological characteristics. The time-lapse system (TLS) has become a promising tool, providing an uninterrupted evaluation of morphological and dynamic parameters of embryo development. Furthermore, TLS sheds light on unknown phenomena such as direct cleavage and incomplete morula compaction. We retrospectively analyzed the morphology (Gardner Score) and morphokinetics (KIDScore) of 835 blastocysts grown in a TLS incubator (Embryoscope+), which were biopsied for preimplantation genetic testing for aneuploidy (PGT-A). Only the embryos that reached the blastocyst stage were included in this study and time-lapse videos were retrospectively reanalysed. According to the pattern of initial cleavages and morula compaction, the embryos were classified as: normal (NC) or abnormal (AC) cleavage, and fully (FCM) or partially compacted (PCM) morulae. No difference was found in early cleavage types or morula compaction patterns between female age groups (< 38, 38-40 and > 40 yo). Most of NC embryos resulted in FCM (≅ 60%), while no embryos with AC resulted in FCM. Aneuploidy rate of AC-PCM group did not differ from that of NC-FCM group in women < 38 yo, but aneuploidy was significantly higher in AC-PCM compared to NC-FCM of women > 40 yo. However, the quality of embryos was lower in AC-PCM blastocysts in women of all age ranges. Morphological and morphokinetic scores declined with increasing age, in the NC-PCM and AC-PCM groups, compared to the NC-FCM. Similar aneuploidy rates among NC-FCM and AC-PCM groups support the hypothesis that PCM in anomalous-cleaved embryos can represent a potential correction mechanism, even though lower morphological/morphokinetic scores are seen on AC-PCM. Therefore, both morphological and morphokinetic assessment should consider these embryonic development phenomena.

Concurrent X chromosome inactivation and upregulation during non-human primate preimplantation development revealed by single-cell RNA-sequencing.

In mammals, dosage compensation of X-linked gene expression between males and females is achieved by inactivation of a single X chromosome in females, while upregulation of the single active X in males and females leads to X:autosome dosage balance. Studies in human embryos revealed that random X chromosome inactivation starts at the preimplantation stage and is not complete by day 12 of development. Alternatively, others proposed that dosage compensation in human preimplantation embryos is achieved by dampening expression from the two X chromosomes in females. Here, we characterize X-linked dosage compensation in another primate, the marmoset (Callithrix jacchus). Analyzing scRNA-seq data from preimplantation embryos, we detected upregulation of XIST at the morula stage, where female embryos presented a significantly higher expression of XIST than males. Moreover, we show an increase of X-linked monoallelically expressed genes in female embryos between the morula and late blastocyst stages, indicative of XCI. Nevertheless, dosage compensation was not achieved by the late blastocyst stage. Finally, we show that X:autosome dosage compensation is achieved at the 8-cell stage, and demonstrate that X chromosome dampening in females does not take place in the marmoset. Our work contributes to the elucidation of primate X-linked dosage compensation.

Aspectos epidemiológicos e hematológicos de cães infectados com Ehrlichia sp e Anaplasma sp em Jataí - GO, Brasil / Epidemiological and hematological aspects of dogs with Ehrlichia sp and Anaplasma sp in Jataí - GO / Aspectos epidemiológicos y hematológicos de perros con Ehrlichia sp y Anaplasma sp en Jataí - GO

No Brasil, as infecções por hemoparasitas representam uma das principais causas de busca por assistência veterinária em cães. Infecções por bactérias da ordem Rickettsiales que incluem Ehrlichia sp. e Anaplasma sp.tem uma grande participação nessas buscas. Foi objetivo deste estudo elucidar aspectos epidemiológicos regionais e hematológicos de infecções por Anaplasma sp.e Ehrlichia sp, muitas vezes assintomáticas, em cães no município de Jataí-GO. Os dados deste estudo foram baseados em amostras de sangue de cães atendidos no Hospital Veterinário da Universidade Federal de Jataí, em Jataí, Goiás, entre os anos de 2005 e 2017. A observação microscópica da presença de mórulas de Ehrlichia sp. e Anaplasma sp. foi utilizada como critério de positividade. 5.015 animais foram atendidos nesse período e 691 (13,78%) deles foram positivos para a presença destes hemoparasitas. Entre esses animais, 9,23% (463/5015) possuíam mórulas de Ehrlichia sp., e 4,37% (219/5015) apresentavam mórulas típicas de Anaplasma sp.,enquanto 0,18% (9/5015) possuíam mórula de ambas as espécies. As fêmeas representaram 57,89% (400/691) dos animais positivos e os machos 42,11% (291/691). 35,02% (242/691) dos animais tinham menos de 1 ano de vida, 54,13% (374/691) tinham entre 1 e 8 anos, e 10,85% (75/691) tinham mais de 8 anos. As fêmeas, com menos de um ano de idade e coinfectadas, apresentaram as alterações mais significativas nos aspectos hematológicos, hematócrito (29%), contagem de hemácias (3 milhões/mm3), contagem de plaquetas (40.000/mm3) e contagem total de leucócitos (16.000 mil/mm3), evidenciando anemia e trombocitopenia. Animais infectados com Anaplasma sp.e Ehrlichia sp.,ao mesmo tempo, apresentaram trombocitopenia mais intensa, evidenciada em machos entre 1 e 8 anos (30.000/mm3). As infecções por Anaplasma sp. causaram uma diminuição severa na média da contagem de glóbulos vermelhos, evidenciada em machos (3 milhões/mm3) e fêmeas com menos de 1 ano (3,79 milhões/mm3).(AU)

In Brazil, infections by hemoparasites represent one of the main causes of seeking veterinary assistance in dogs. Infections with bacteria of the order Rickettsiales that include Ehrlichia sp. and Anaplasma sp. has a great participation in these assistances. The objective of this study was to elucidate regional epidemiologic and hematologic aspects of infections by Anaplasma sp. and Ehrlichia sp.,often asymptomatic, in dogs in the municipality of Jataí-GO. The data in this study were based on blood samples from dogs treated at the Veterinary Hospital of the Federal University of Jataí, in Jataí, Goiás, between the years 2005 and 2017. Microscopic observation of the presence of Ehrlichia sp. and Anaplasma sp. was used as a positivity criterion. 5,015 animals were treated during this period and 691 (13.78%) of them were positive for the presence of these hemoparasites. Among these animals, 9.23% (463/5015) had morulae from Ehrlichia sp., and 4.37% (219/5015) had morulae typical of Anaplasma sp., while 0.18% (9/5015) had morulae of both species. Females represented 57.89% (400/691) of positive animals and males 42.11% (291/691). 35.02% (242/691) of the animals were less than one year old, 54.13% (374/691) were between 1 and 8 years old, and 10.85% (75/691) were over 8 years old. Females, less than one year old and co-infected, showed the most significant changes in hematological, hematocrit (29%), red blood cell count (3 million / mm3), platelet count (40,000 / mm3) and total blood count leukocytes (16,000 thousand / mm3), showing anemia and thrombocytopenia. Animals infected with Anaplasma sp. and Ehrlichia sp., at the same time, presented more intense thrombocytopenia, evidenced in males between 1 and 8 years old (30,000 / mm3). Infections by Anaplasma sp. caused a severe decrease in the mean red blood cell count, evidenced in males (3 million / mm3) and females under 1 year (3.79 million / mm3).(AU)

En Brasil, las infecciones por hemoparásitos representan una de las principales causas de búsqueda de asistencia veterinaria en perros. Infecciones por bacterias del orden Rickettsiales que incluyen Ehrlichia sp.y Anaplasma sp.tienen una gran participación en estas búsquedas. El objetivo de este estudio fue dilucidar los aspectos epidemiológicos regionales y hematológicos de las infecciones por Anaplasma sp.y Ehrlichia sp., muchas veces asintomáticas, en perros del municipio de Jataí-GO. Los datos de este estudio se basaron en muestras de sangre de perros tratados en el Hospital Veterinario de la Universidad Federal de Jataí, en Jataí, Goiás, entre los años 2005 y 2017. Observación microscópica de la presencia de Ehrlichia sp. y Anaplasma sp. se utilizó como criterio de positividad. Durante este período se trataron 5.015 animales y 691 (13,78%) de ellos resultaron positivos para la presencia de estos hemoparásitos. Entre estos animales, 9,23% (463/5015) tenía mórulas de Ehrlichia sp., y 4,37% (219/5015) tenían mórulas típicas de Anaplasma sp., mientras que 0,18% (9/5015) tenían mórulas de ambas especies. Las hembras representaron 57,89% (400/691) de los animales positivos y los machos 42,11% (291/691). 35,02% (242/691) de los animales tenían menos de 1 año, 54,13% (374/691) tenían entre 1 y 8 años y 10,85% (75/691) tenían más de 8 años. Las hembras, menores de un año y coinfectadas, mostraron los cambios más significativos en hematología, hematocrito (29%), recuento de glóbulos rojos (3 millones/mm3), recuento de plaquetas (40.000/mm3) y recuento sanguíneo total leucocitos (16.000 mil/mm3), mostrando anemia y trombocitopenia. Los animales infectados por Anaplasma sp. y Ehrlichia sp., al mismo tiempo, presentaron trombocitopenia más intensa, evidenciada en machos entre 1 y 8 años (30.000 /mm3). Infecciones por Anaplasma sp. causó una disminución severa en el recuento promedio de glóbulos rojos, evidenciada en machos (3 millones/mm3) y hembras menores de 1 año (3.79 millones/mm3).(AU)

Aspectos epidemiológicos e hematológicos de cães infectados com Ehrlichia sp e Anaplasma sp em Jataí - GO, Brasil / Epidemiological and hematological aspects of dogs with Ehrlichia sp and Anaplasma sp in Jataí - GO / Aspectos epidemiológicos y hematológicos de perros con Ehrlichia sp y Anaplasma sp en Jataí - GO

No Brasil, as infecções por hemoparasitas representam uma das principais causas de busca por assistência veterinária em cães. Infecções por bactérias da ordem Rickettsiales que incluem Ehrlichia sp. e Anaplasma sp.tem uma grande participação nessas buscas. Foi objetivo deste estudo elucidar aspectos epidemiológicos regionais e hematológicos de infecções por Anaplasma sp.e Ehrlichia sp, muitas vezes assintomáticas, em cães no município de Jataí-GO. Os dados deste estudo foram baseados em amostras de sangue de cães atendidos no Hospital Veterinário da Universidade Federal de Jataí, em Jataí, Goiás, entre os anos de 2005 e 2017. A observação microscópica da presença de mórulas de Ehrlichia sp. e Anaplasma sp. foi utilizada como critério de positividade. 5.015 animais foram atendidos nesse período e 691 (13,78%) deles foram positivos para a presença destes hemoparasitas. Entre esses animais, 9,23% (463/5015) possuíam mórulas de Ehrlichia sp., e 4,37% (219/5015) apresentavam mórulas típicas de Anaplasma sp.,enquanto 0,18% (9/5015) possuíam mórula de ambas as espécies. As fêmeas representaram 57,89% (400/691) dos animais positivos e os machos 42,11% (291/691). 35,02% (242/691) dos animais tinham menos de 1 ano de vida, 54,13% (374/691) tinham entre 1 e 8 anos, e 10,85% (75/691) tinham mais de 8 anos. As fêmeas, com menos de um ano de idade e coinfectadas, apresentaram as alterações mais significativas nos aspectos hematológicos, hematócrito (29%), contagem de hemácias (3 milhões/mm3), contagem de plaquetas (40.000/mm3) e contagem total de leucócitos (16.000 mil/mm3), evidenciando anemia e trombocitopenia. Animais infectados com Anaplasma sp.e Ehrlichia sp.,ao mesmo tempo, apresentaram trombocitopenia mais intensa, evidenciada em machos entre 1 e 8 anos (30.000/mm3). As infecções por Anaplasma sp. causaram uma diminuição severa na média da contagem de glóbulos vermelhos, evidenciada em machos (3 milhões/mm3) e fêmeas com menos de 1 ano (3,79 milhões/mm3).

In Brazil, infections by hemoparasites represent one of the main causes of seeking veterinary assistance in dogs. Infections with bacteria of the order Rickettsiales that include Ehrlichia sp. and Anaplasma sp. has a great participation in these assistances. The objective of this study was to elucidate regional epidemiologic and hematologic aspects of infections by Anaplasma sp. and Ehrlichia sp.,often asymptomatic, in dogs in the municipality of Jataí-GO. The data in this study were based on blood samples from dogs treated at the Veterinary Hospital of the Federal University of Jataí, in Jataí, Goiás, between the years 2005 and 2017. Microscopic observation of the presence of Ehrlichia sp. and Anaplasma sp. was used as a positivity criterion. 5,015 animals were treated during this period and 691 (13.78%) of them were positive for the presence of these hemoparasites. Among these animals, 9.23% (463/5015) had morulae from Ehrlichia sp., and 4.37% (219/5015) had morulae typical of Anaplasma sp., while 0.18% (9/5015) had morulae of both species. Females represented 57.89% (400/691) of positive animals and males 42.11% (291/691). 35.02% (242/691) of the animals were less than one year old, 54.13% (374/691) were between 1 and 8 years old, and 10.85% (75/691) were over 8 years old. Females, less than one year old and co-infected, showed the most significant changes in hematological, hematocrit (29%), red blood cell count (3 million / mm3), platelet count (40,000 / mm3) and total blood count leukocytes (16,000 thousand / mm3), showing anemia and thrombocytopenia. Animals infected with Anaplasma sp. and Ehrlichia sp., at the same time, presented more intense thrombocytopenia, evidenced in males between 1 and 8 years old (30,000 / mm3). Infections by Anaplasma sp. caused a severe decrease in the mean red blood cell count, evidenced in males (3 million / mm3) and females under 1 year (3.79 million / mm3).

En Brasil, las infecciones por hemoparásitos representan una de las principales causas de búsqueda de asistencia veterinaria en perros. Infecciones por bacterias del orden Rickettsiales que incluyen Ehrlichia sp.y Anaplasma sp.tienen una gran participación en estas búsquedas. El objetivo de este estudio fue dilucidar los aspectos epidemiológicos regionales y hematológicos de las infecciones por Anaplasma sp.y Ehrlichia sp., muchas veces asintomáticas, en perros del municipio de Jataí-GO. Los datos de este estudio se basaron en muestras de sangre de perros tratados en el Hospital Veterinario de la Universidad Federal de Jataí, en Jataí, Goiás, entre los años 2005 y 2017. Observación microscópica de la presencia de Ehrlichia sp. y Anaplasma sp. se utilizó como criterio de positividad. Durante este período se trataron 5.015 animales y 691 (13,78%) de ellos resultaron positivos para la presencia de estos hemoparásitos. Entre estos animales, 9,23% (463/5015) tenía mórulas de Ehrlichia sp., y 4,37% (219/5015) tenían mórulas típicas de Anaplasma sp., mientras que 0,18% (9/5015) tenían mórulas de ambas especies. Las hembras representaron 57,89% (400/691) de los animales positivos y los machos 42,11% (291/691). 35,02% (242/691) de los animales tenían menos de 1 año, 54,13% (374/691) tenían entre 1 y 8 años y 10,85% (75/691) tenían más de 8 años. Las hembras, menores de un año y coinfectadas, mostraron los cambios más significativos en hematología, hematocrito (29%), recuento de glóbulos rojos (3 millones/mm3), recuento de plaquetas (40.000/mm3) y recuento sanguíneo total leucocitos (16.000 mil/mm3), mostrando anemia y trombocitopenia. Los animales infectados por Anaplasma sp. y Ehrlichia sp., al mismo tiempo, presentaron trombocitopenia más intensa, evidenciada en machos entre 1 y 8 años (30.000 /mm3). Infecciones por Anaplasma sp. causó una disminución severa en el recuento promedio de glóbulos rojos, evidenciada en machos (3 millones/mm3) y hembras menores de 1 año (3.79 millones/mm3).

In Vitro Development and Mitochondrial Gene Expression in Brown Brocket Deer (Mazama gouazoubira) Embryos Obtained by Interspecific Somatic Cell Nuclear Transfer.

The genetic diversity of Neotropical deer is increasingly jeopardized, owing to declining population size. Thus, the formation of cryobanking of somatic cells is important for the preservation of these species using cloning. The transformation of these cells into viable embryos has been hampered by a lack of endangered species oocytes. Accordingly, the aim of this study was to produce brown brocket deer embryos by interspecific somatic cell nuclear transfer (iSCNT), using goat or cattle oocytes as cytoplasts, and to elucidate embryo mitochondrial activity by measuring the expression levels of ATP6, COX3, and ND5. Cattle embryos produced by in vitro fertilization (IVF) were used as a control. There were no differences in the development of embryos produced by traditional SCNT and iSCNT when using either the goat cytoplasts (38.4% vs. 25.0% cleaved and 40.0% vs. 50.0% morula rates, respectively) or cattle cytoplast (72.8% vs. 65.5% cleaved and 11.3% vs. 5.9% blastocyst rates, respectively). Concerning the gene expression, no significant difference was observed when goat oocytes were used as cytoplasts. However, when using cattle oocytes and 16S as a reference gene, the iSCNT upregulated COX3, when compared with SCNT group. In contrast, when GAPDH was used as a reference gene, all the evaluated genes were upregulated in the iSCNT group, when compared with the IVF group. When compared with the SCNT group, only the expression of ATP6 was statistically different. In conclusion, it was demonstrated that interspecific nuclear transfer is a potentially useful tool for conservation programs of endangered similar deer species.

Sleep deprivation decreases the reproductive capacity by affecting the arrival of morulas in the uterus.

A previous animal study by our group found that sleep deprivation during preimplantation was associated with decreased pregnancy maintenance. Given its impact on human society, we aimed in the current study to assess whether sleep deprivation affects blastocyst gene expression and/or the implantation process. For this, pregnant mice (gestational day 0 [GD 0]) were assigned into paradoxical sleep deprivation (SD, 72 hr; multiple platform method) and, a control (CT) group. Animals were euthanized on GD 3.5 and blood, uterus (embryos) and fallopian tube were collected. Then, 89% of CT presented blastocysts in the uterus versus 25% from SD group. Compared to CT, SD presented lighter relative uterus weight, increased plasma concentrations of corticosterone and testosterone, decreased concentrations of progesterone and luteinizing hormone, but no statistical differences in plasma concentrations of 17ß-estradiol and follicle stimulating hormone. There were no differences in uterus and blastocyst gene expression related to embryo implantation and development, and no alteration in blastocysts global DNA methylation. Considering this, the decreased pregnancy maintenance after sleep deprivation seems not to be associated with implantation losses or developmental problems related to the blastocysts. It is likely that complications in morula development and/or its movement through the fallopian tubes affect the pregnancy rate, since only 25% of SD females presented a blastocyst on the GD 3.5. In fact, three out of four females without blastocysts in the uterus presented morula in the fallopian tubes due to a phase delay. Additionally, we suggest that the observed hormonal changes may play a role in this outcome.

Pregnancy rate after fixed-time transfer of cryopreserved embryos collected by non-surgical route in Lacaune sheep.

This study investigated the feasibility of applying fixed-time (cryopreserved) embryo transfer in ewes. Embryos (n = 106) were non-surgically recovered from superovulated donors (n = 39) on day 6-7 after oestrus. Straws containing one or two embryos (morulae and/or blastocysts) subjected to either slow freezing (SF, n = 62) or vitrification (VT, n = 44) were randomly used within fixed-time embryo transfer on Day 8.5. Recipient ewes were nulliparous (n = 58) bearing corpora lutea after synchronous oestrous induction protocol. The pregnancy rate was higher (p = .03) in SF (39.4%) than VT (16.9%) and survival rate tended (p = .08) to be higher in SF than in VT (25.8% vs. 15.9%). Lambing rates were similar (p = .13) between SF (20.9%) and VT (15.9%). Embryos recovered by non-surgical route after cervical dilation treatment and later cryopreserved by either slow freezing or vitrification produced reasonable pregnancy rates after FTET.

Embryonic development in Zungaro jahu.

The aim of this study was to characterize the embryonic development of Zungaro jahu, a fresh water teleostei commonly known as 'jaú'. Samples were collected at pre-determined times from oocyte release to larval hatching and analysed under light microscopy, transmission electron microscopy and scanning electron microscopy. At the first collection times, the oocytes and eggs were spherical and yellowish, with an evident micropyle. Embryo development took place at 29.4 ± 1.5°C and was divided into seven stages: zygote, cleavage, morula, blastula, gastrula, organogenesis, and hatching. The differentiation of the animal and vegetative poles occured during the zygote stage, at 10 min post-fertilization (mpf), leading to the development of the egg cell at 15 mpf. From 20 to 75 mpf, successive cleavages resulted in the formation of 2, 4, 8, 16, 32 and 64 blastomeres. The morula stage was observed between 90 and 105 mpf, and the blastula and gastrula stage at 120 and 180 mpf; respectively. The end of the gastrula stage was characterized by the presence of the yolk plug at 360 mpf. Organogenesis followed, with differentiation of the cephalic and caudal regions, elongation of the embryo by the cephalo-caudal axis, and somitogenesis. Hatching occurred at 780 mpf, with mean larval total length of 3.79 ± 0.11 mm.

Effect of DNase treatment on RNA extraction from preimplantation murine embryos.

The quality of RNA is crucial when performing microarray experiments. This is particularly important when dealing with preimplantation embryos, from which a minimum yield of RNA of good quality can be produced. We report the optimization of several RNA extraction methods applied to preimplantation embryos at different stages of development. The quality of the samples was confirmed using a microarray and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis. A total of 30 cultured two-cell stage embryos of ICR mice were pooled at the 8-cell, morula, and blastocyst stages. The embryos were divided into two groups comprising DNase-treated and non-DNase-treated RNA samples. Total RNA was extracted using a Pico Pure RNA Isolation Kit following the manufacturer protocol, with some modifications. Lysed samples were bound to a silica-based filter, treated with deoxyribonuclease I (DNase I), and washed several times before elution. RNA concentration and integrity were evaluated using an Agilent 2100 Bioanalyzer and an RNA 6000 Pico Assay kit. Although concentrations of non-DNase-treated RNAs were higher than DNase-treated RNA, DNase-treated RNA gave a higher RNA integrity number compared with non-DNase-treated RNA. Inclusion of DNase treatment in the RNA extraction procedure gave the best quality RNA samples from preimplantation embryos, as validated by microarray and RT-qPCR quality control.

Supplementation of fetal bovine serum alters histone modification H3R26me2 during preimplantation development of in vitro produced bovine embryos / Suplementação com soro fetal bovino altera a modificação de histona H3R26me2 durante o período pré-implantacional em embriões bovinos produzidos in vitro

In vitro production (IVP) of bovine embryos is not only of great economic importance to the cattle industry, but is also an important model for studying embryo development. The aim of this study was to evaluate the histone modification, H3R26me2 during pre-implantation development of IVP bovine embryos cultured with or without serum supplementation and how these in vitro treatments compared to in vivo embryos at the morula stage. After in vitro maturation and fertilization, bovine embryos were cultured with either 0 or 2.5% fetal bovine serum (FBS). Development was evaluated and embryos were collected and fixed at different stages during development (2-, 4-, 8-, 16-cell, morula and blastocyst). Fixed embryos were then used for immunofluorescence utilizing an antibody for H3R26me2. Images of stained embryos were analyzed as a percentage of total DNA. Embryos cultured with 2.5% FBS developed to blastocysts at a greater rate than 0%FBS groups (34.85±5.43% vs. 23.38±2.93%; P<0.05). Levels of H3R26me2 changed for both groups over development. In the 0%FBS group, the greatest amount of H3R26me2 staining was at the 4-cell (P<0.05), 16-cell (P<0.05) and morula (P<0.05) stages. In the 2.5%FBS group, only 4-cell stage embryos were significantly higher than all other stages (P<0.01). Morula stage in vivo embryos had similar levels as the 0%FBS group, and both were significantly higher than the 2.5%FBS group. These results suggest that the histone modification H3R26me2 is regulated during development of pre-implantation bovine embryos, and that culture conditions greatly alter this regulation...

A produção in vitro (PIV) de embriões de bovinos não é apenas de grande importância econômica para a pecuária, mas é também um importante modelo para estudar o desenvolvimento embrionário. O objetivo deste estudo foi avaliar a modificação de histona, H3R26me2 durante o desenvolvimento pré-implantacional em embriões bovinos produzidos in vitro, cultivados com ou sem suplementação de soro fetal bovino (SFB), bem como comparar essa modificação específica entre mórulas produzidas in vitro e in vivo. Após a maturação in vitro e fertilização, embriões foram cultivados com suplementação de 0 ou 2,5% SFB. O desenvolvimento embrionário foi avaliado e embriões foram coletados e fixados em diferentes fases durante o desenvolvimento (2, 4, 8 e 16 células, mórula e blastocisto). Os embriões fixados foram avaliados por imunofluorescência utilizando um anticorpo para H3R26me2. Imagens de embriões corados foram analisadas baseadas na porcentagem do DNA total. Embriões cultivados com 2,5% SFB tiveram uma taxa de desenvolvimento ao estágio de blastocisto maior que o grupo que não recebeu suplementação com SFB (34.85±5,43% vs 23.38±,93%; P<0,05). Níveis de H3R26me2 variaram para ambos os grupos ao longo do desenvolvimento. No grupo 0% SFB, a marcação para H3R26me2 foi mais intensa nos estágios de 4 células (P<0,05), 16 células (P<0,05) e mórula (P<0.05). No grupo 2.5% SFB, apenas os embriões de 4 células tiveram marcação significativamente maior que todas as outras fases (P<0,01). Mórulas produzidas in vivo apresentaram níveis de H3R26me2 semelhantes ao grupo 0% SFB, e ambos foram significativamente maiores que o grupo 2.5% SFB. Estes resultados sugerem que a modificação de histona H3R26me2 é regulada durante o desenvolvimento pré-implantacional de embriões bovinos, e que as condições de cultura alteram de maneira importante esta regulação...

Supplementation of fetal bovine serum alters histone modification H3R26me2 during preimplantation development of in vitro produced bovine embryos / Suplementação com soro fetal bovino altera a modificação de histona H3R26me2 durante o período pré-implantacional em embriões bovinos produzidos in vitro

In vitro production (IVP) of bovine embryos is not only of great economic importance to the cattle industry, but is also an important model for studying embryo development. The aim of this study was to evaluate the histone modification, H3R26me2 during pre-implantation development of IVP bovine embryos cultured with or without serum supplementation and how these in vitro treatments compared to in vivo embryos at the morula stage. After in vitro maturation and fertilization, bovine embryos were cultured with either 0 or 2.5% fetal bovine serum (FBS). Development was evaluated and embryos were collected and fixed at different stages during development (2-, 4-, 8-, 16-cell, morula and blastocyst). Fixed embryos were then used for immunofluorescence utilizing an antibody for H3R26me2. Images of stained embryos were analyzed as a percentage of total DNA. Embryos cultured with 2.5% FBS developed to blastocysts at a greater rate than 0%FBS groups (34.85±5.43% vs. 23.38±2.93%; P<0.05). Levels of H3R26me2 changed for both groups over development. In the 0%FBS group, the greatest amount of H3R26me2 staining was at the 4-cell (P<0.05), 16-cell (P<0.05) and morula (P<0.05) stages. In the 2.5%FBS group, only 4-cell stage embryos were significantly higher than all other stages (P<0.01). Morula stage in vivo embryos had similar levels as the 0%FBS group, and both were significantly higher than the 2.5%FBS group. These results suggest that the histone modification H3R26me2 is regulated during development of pre-implantation bovine embryos, and that culture conditions greatly alter this regulation.(AU)

A produção in vitro (PIV) de embriões de bovinos não é apenas de grande importância econômica para a pecuária, mas é também um importante modelo para estudar o desenvolvimento embrionário. O objetivo deste estudo foi avaliar a modificação de histona, H3R26me2 durante o desenvolvimento pré-implantacional em embriões bovinos produzidos in vitro, cultivados com ou sem suplementação de soro fetal bovino (SFB), bem como comparar essa modificação específica entre mórulas produzidas in vitro e in vivo. Após a maturação in vitro e fertilização, embriões foram cultivados com suplementação de 0 ou 2,5% SFB. O desenvolvimento embrionário foi avaliado e embriões foram coletados e fixados em diferentes fases durante o desenvolvimento (2, 4, 8 e 16 células, mórula e blastocisto). Os embriões fixados foram avaliados por imunofluorescência utilizando um anticorpo para H3R26me2. Imagens de embriões corados foram analisadas baseadas na porcentagem do DNA total. Embriões cultivados com 2,5% SFB tiveram uma taxa de desenvolvimento ao estágio de blastocisto maior que o grupo que não recebeu suplementação com SFB (34.85±5,43% vs 23.38±,93%; P<0,05). Níveis de H3R26me2 variaram para ambos os grupos ao longo do desenvolvimento. No grupo 0% SFB, a marcação para H3R26me2 foi mais intensa nos estágios de 4 células (P<0,05), 16 células (P<0,05) e mórula (P<0.05). No grupo 2.5% SFB, apenas os embriões de 4 células tiveram marcação significativamente maior que todas as outras fases (P<0,01). Mórulas produzidas in vivo apresentaram níveis de H3R26me2 semelhantes ao grupo 0% SFB, e ambos foram significativamente maiores que o grupo 2.5% SFB. Estes resultados sugerem que a modificação de histona H3R26me2 é regulada durante o desenvolvimento pré-implantacional de embriões bovinos, e que as condições de cultura alteram de maneira importante esta regulação.(AU)

Structural analysis of embryogenesis of Leiarius marmoratus (Siluriformes: Pimelodidae).

Embryological studies in fish species are useful to the understanding of their biology and systematics. The available biological data in Leiarius marmoratus are scarce and additional information about its reproductive biology is needed, mainly because this species has been commercially exploited and used in production of hybrid lineages. In order to evaluate the temporal-morphological embryonic modifications in L. marmoratus, samples of nearly 200 embryos were collected at random at different stages of development, starting from fecundation (time zero). Embryos were fixed in modified Karnovsk's solution and 2.5% glutaraldehyde, processed and analysed under optic and electron microscopy. The incubation period of L. marmoratus was equal to 14.42 h at a mean temperature of 28.3 ± 0.07°C. The following stages of embryonic development were established: zygote, cleavage, gastrula, organogenesis and hatching. These stages were divided into phases, as follows: cleavage - phases of 2, 4, 8, 16, 32 and 64 cells and morula; gastrula - phases of 25, 50, 75 and 90% of epiboly and blastopore closure; and organogenesis - neurula, segmentation and pre-larval phases. The embryogenesis of L. marmoratus was typical of neotropical teleosteans, with peculiarities in species development.

Cryotolerance of Day 2 or Day 6 in vitro produced ovine embryos after vitrification by Cryotop or Spatula methods.

This study was conducted to evaluate the cryotolerance of in vitro produced ovine embryos submitted to vitrification at different developmental stages using two methods of minimum volume and rapid cooling rate. Embryos were vitrified at early stage (2 to 8-cells) on Day 2 or at advanced stage (morulae and blastocysts) on Day 6 after in vitro fertilization. Vitrification procedure consisted of the Cryotop (Day 2, n=165; Day 6, n=174) or the Spatula method (Day 2, n=165; Day 6, n=175). Non vitrified embryos were maintained in in vitro culture as a control group (n=408). Embryo survival was determined at 3h and 24h after warming, development and hatching rates were evaluated on Day 6 and Day 8 after fertilization, and total cell number was determined on expanded blastocysts. Embryo survival at 24h after warming increased as the developmental stage progressed (P<0.05) and was not affected by the vitrification method. The ability for hatching of survived embryos was not affected by the stage of the embryos at vitrification or by the vitrification method. Thus, the proportion of hatching from vitrified embryos was determined by the survival rate and was lower for Day 2 than Day 6 vitrified embryos. The percentage of blastocysts on Day 8 was lower for the embryos vitrified on Day 2 than Day 6 (P<0.05), and was lower for both days of vitrification than for non-vitrified embryos (P<0.05). No interaction of embryo stage by vitrification method was found (P=NS) and no significant difference was found in the blastocyst cell number among vitrified and non-vitrified embryos. In conclusion, both methods using minimum volume and ultra-rapid cooling rate allow acceptable survival and development rates in Day 2 and Day 6 in vitro produced embryos in sheep. Even though early stage embryos showed lower cryotolerance, those embryos that survive the vitrification-warming process show high development and hatching rates, similar to vitrification of morulae or blastocysts.

Expression pattern of JMJD1C in oocytes and its impact on early embryonic development.

Cell reprogramming mediated by histone methylation and demethylation is crucial for the activation of the embryonic genome in early embryonic development. In this study, we employed quantitative real-time polymerase chain reaction (qRT-PCR) to detect mRNA levels and expression patterns of all known histone demethylases in early germinal vesicle stage and in vitro-matured metaphase II (MII) oocytes (which are commonly used as donor cells for nuclear transfer). On screening, the Jumonji domain containing 1C (JMJD1C) gene had the highest level of expression and hence was used for subsequent experiments. We also found that JMJD1C was primarily expressed in the nucleus and showed relatively high levels of expression at the 2-cell, 4-cell, 8-cell, 16-cell, morula, and blastocyst stages of embryos developed from MII oocytes fertilized in vitro. Further, we knocked down the JMJD1C gene in MII oocytes using siRNA and monitored the cleavage of zygotes and development of early embryos after in vitro fertilization. The results showed that the zygote cleavage and blastocyst rates of the transfection group were reduced by 57.1 ± 0.07 and 50 ± 0.01% respectively, which were significantly lower than those of the negative control group (P < 0.05). These data suggest that JMJD1C plays a key role in the normal development of early bovine embryos. Our results also provide a theoretical basis for the investigation of the role and molecular mechanism of histone demethylation in the early development of bovine embryos.

H3K27me3 may be associated with Oct4 and Sox2 in mouse preimplantation embryos.

As a core member of polycomb repressive complex 2, the transcription and enzyme activity of enhancer of zeste homolog 2 (Ezh2) is directly involved in the trimethylation of lysine 27 on histone H3. In this study, the fluorescence intensity of H3K27me3 in mouse in vivo morulae and blastocysts was compared by indirect immunofluorescence staining. We found that demethylation of H3K27me3 occurred during the blastocyst stage. Real-time polymerase chain reaction was performed to investigate Ezh2 expression in oocytes and in preimplantation embryos. Ezh2 expression peaked during the zygote stage and gradually decreased from the 2-cell stage, exhibiting an inverse pattern when compared with Oct4 and Sox2 mRNA in mouse preimplantation embryos. To understand the role of development-related genes on the transcription of mouse Ezh2, a promoter assay was performed in NIH/3T3 cells. Ezh2 expression was markedly suppressed by Oct4 and Sox2 alone in a dose-dependent manner, while Ezh2 promoter activity in co-transfection with Nanog, Klf-4, and c-Myc groups showed no significant change as compared with the control. Our data suggest that the demethylation of H3K27me3 is caused by the degressive expression and activity of Ezh2 in blastocysts, leading to increased expression of developmentally important transcription factors. We also observed negative effects of Oct4 and Sox2 on the transcription of Ezh2 and identified Oct4 and Sox2 as novel negative regulators of Ezh2 at the post-translation level in a mouse preimplantation embryo.

The seasonal and ovarian status effects on in vitro production of domestic cat embryos between Equator and Tropic of Capricorn / Efeitos da sazonalidade e da condição ovariana sobre a produção in vitro de embriões de gatos domésticos em uma região entre o Equador e o Trópico de Capricórnio

From the Tropic of Capricorn to Equator, the seasonality of domestic cat is known to be absent, i.e., these animals are considered non-seasonal breeders at these regions. We hypothesized that this particularity might have some influence on in vitro embryo production. The aim of this experiment was to determine the percentage of cleavage and morulae and blastocyst formation produced from oocytes recovered from queen ovaries of three distinct status - follicular, luteal or inactive - during two different reproductive seasons experienced by cats in southeast of Brazil (22°53'09" S and 48°26'42" W) - non breeding season (NBS), comprehending January to March; and breeding season (BS), August to October. Thirty queens were neutered. [...] During NBS, from a total of 272 (inactive), 162 (luteal) and 134 (follicular) fertilized oocytes, the percentage of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 24.63, 16.54 and 8.09 respectively; for those derived from luteal ovaries, the percentage was 21.6, 12.96 and 8.64, and for those from follicular ovaries, they were 24.62, 16.41 and 8.21. Considering BS, from a total of 102 (inactive), 198 (luteal) and 86 (follicular) fertilized oocytes, the relative frequency (%) of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 64.7, 41.17 and 23.53 respectively; for those derived from luteal ovaries, the percentage was 64.14, 40.41 and 23.73, and for those from follicular ovaries, they were 63.95, 39.54 and 24.41. The results of this experiment demonstrate that no statistically significant difference (P<0.05) was verified in the frequency of cleaved embryos and morulae and blastocyst formation when comparing the three ovarian conditions in the same season. However the breeding season presented better results considering cleavage and morulae and blastocyst formation.

Do Trópico de Capricórnio ao Equador, sabe-se que a sazonalidade no gato domestico é ausente, i.e., estes animais são considerados reprodutores não sazonais nestas regiões. [...] O objetivo deste experimento foi determinar a porcentagem de clivagem e formação de mórulas e blastocistos produzidos a partir de oócitos recuperados de ovários de gatas em três condições - folicular, lútea ou inativa - durante duas estações reprodutivas pelas quais gatas passam na região sudeste do Brasil (22°53'09" S e 48°26'42" O) - estação não reprodutiva (ENR), que compreende os meses de janeiro a março; e estação reprodutiva (ER), agosto à outubro. Trinta gatas foram castradas. [...] Durante a ENR, de um total de 272 (inativo), 162 (lútea) e 134 (folicular) oócitos fertilizados, a porcentagem de clivagem de zigotos, formação de mórulas e de blastocistos derivados de ovários inativos foi 24,63, 16,54 e 8,09 respectivamente; para aqueles oriundos de ovários na condição lútea, a porcentagem foi de 21,6, 12,96 e 8,64, e para aqueles provenientes de ovários na fase folicular, foi de 24,62, 16,41 e 8,21. Considerando a ER, de um total de 102 (inativo), 198 (lútea) e 86 (folicular) oócitos fertilizados, a frequência relativa (%) de zigotos clivados, mórulas e blastocistos derivados de ovários na condição inativa foi de 64,7, 41,17 e 23,53 respectivamente; para aqueles oriundos de ovários na condição lútea, a porcentagem foi de 64,14, 40,41 e 23,73, e para aqueles provenientes de ovários na fase folicular, foi de 63,95, 39,54 e 24,41. Os resultados deste experimento demonstraram que não houve diferença estatística significante (P < 0.05) na frequência de embriões clivados e na formação de mórulas e blastocistos quando comparadas as três condições ovarianas dentro da mesma estação. Entretanto, a ER apresentou resultados melhores considerando as taxas de clivagem e formação de mórula e de blastocisto se comparada à ENR.

The seasonal and ovarian status effects on in vitro production of domestic cat embryos between Equator and Tropic of Capricorn / Efeitos da sazonalidade e da condição ovariana sobre a produção in vitro de embriões de gatos domésticos em uma região entre o Equador e o Trópico de Capricórnio

From the Tropic of Capricorn to Equator, the seasonality of domestic cat is known to be absent, i.e., these animals are considered non-seasonal breeders at these regions. We hypothesized that this particularity might have some influence on in vitro embryo production. The aim of this experiment was to determine the percentage of cleavage and morulae and blastocyst formation produced from oocytes recovered from queen ovaries of three distinct status - follicular, luteal or inactive - during two different reproductive seasons experienced by cats in southeast of Brazil (22°53'09" S and 48°26'42" W) - non breeding season (NBS), comprehending January to March; and breeding season (BS), August to October. Thirty queens were neutered. [...] During NBS, from a total of 272 (inactive), 162 (luteal) and 134 (follicular) fertilized oocytes, the percentage of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 24.63, 16.54 and 8.09 respectively; for those derived from luteal ovaries, the percentage was 21.6, 12.96 and 8.64, and for those from follicular ovaries, they were 24.62, 16.41 and 8.21. Considering BS, from a total of 102 (inactive), 198 (luteal) and 86 (follicular) fertilized oocytes, the relative frequency (%) of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 64.7, 41.17 and 23.53 respectively; for those derived from luteal ovaries, the percentage was 64.14, 40.41 and 23.73, and for those from follicular ovaries, they were 63.95, 39.54 and 24.41. The results of this experiment demonstrate that no statistically significant difference (P<0.05) was verified in the frequency of cleaved embryos and morulae and blastocyst formation when comparing the three ovarian conditions in the same season. However the breeding season presented better results considering cleavage and morulae and blastocyst formation.(AU)

Do Trópico de Capricórnio ao Equador, sabe-se que a sazonalidade no gato domestico é ausente, i.e., estes animais são considerados reprodutores não sazonais nestas regiões. [...] O objetivo deste experimento foi determinar a porcentagem de clivagem e formação de mórulas e blastocistos produzidos a partir de oócitos recuperados de ovários de gatas em três condições - folicular, lútea ou inativa - durante duas estações reprodutivas pelas quais gatas passam na região sudeste do Brasil (22°53'09" S e 48°26'42" O) - estação não reprodutiva (ENR), que compreende os meses de janeiro a março; e estação reprodutiva (ER), agosto à outubro. Trinta gatas foram castradas. [...] Durante a ENR, de um total de 272 (inativo), 162 (lútea) e 134 (folicular) oócitos fertilizados, a porcentagem de clivagem de zigotos, formação de mórulas e de blastocistos derivados de ovários inativos foi 24,63, 16,54 e 8,09 respectivamente; para aqueles oriundos de ovários na condição lútea, a porcentagem foi de 21,6, 12,96 e 8,64, e para aqueles provenientes de ovários na fase folicular, foi de 24,62, 16,41 e 8,21. Considerando a ER, de um total de 102 (inativo), 198 (lútea) e 86 (folicular) oócitos fertilizados, a frequência relativa (%) de zigotos clivados, mórulas e blastocistos derivados de ovários na condição inativa foi de 64,7, 41,17 e 23,53 respectivamente; para aqueles oriundos de ovários na condição lútea, a porcentagem foi de 64,14, 40,41 e 23,73, e para aqueles provenientes de ovários na fase folicular, foi de 63,95, 39,54 e 24,41. Os resultados deste experimento demonstraram que não houve diferença estatística significante (P < 0.05) na frequência de embriões clivados e na formação de mórulas e blastocistos quando comparadas as três condições ovarianas dentro da mesma estação. Entretanto, a ER apresentou resultados melhores considerando as taxas de clivagem e formação de mórula e de blastocisto se comparada à ENR.(AU)

In vitro development of murine embryos in presence of campylobacter fetus / Desarrollo de embriones murinos in vitro en presencia de campylobacter fetus

Bovine campylobacteriosis caused by Campylobacter fetus is associated with reproductive losses. The knowledge about the mechanisms of bacterial pathogenesis is limited, then a murine experimental model is proposed. BALB/c females and males were used. Two-cell embryos were cultured in Ham-F10 as control group (CG). Treatment groups were constituted by the addition of Cfv 1 and 3, or Cff 2 and 5. Morulae were placed in Ham-F10 (CG); treatment groups were constituted by the addition of Cfv27, CFF (cell-free filtrate) and Brucella broth (BB). Blastocysts were cultured in MEM (CG); challenge group were constituted by the addition of Cfv 27. Differentiation, hatching, hatched, adhesion and expansion were evaluated. Results were analyzed by Chi2 test. In two-cell embryo, the differentiation rate was not modified when the study strains were added (p > 0.05). The differentiation rate at 24 h for embryos at the morula stage was lower for BB, Cfv, and CFF, compared with CG (p < 0.05). After 48 h culture, no differences were observed in blastocyst formation for Cfv and BB, compared to CG (p > 0.05). However, the differentiation rate for the CFF group was lower than for CG (p < 0.05). At 48 and 72 h, the hatching rate was higher in CFF and Cfv groups than in CG (p < 0.05). Differences were not detected in blastocyst cultures. In conclusion, under these experimental conditions, Cf was not detrimental to the development of murine embryos. Efforts will be intensified to establish in vitro infection models that reproduce their pathogenicity.

La campilobacteriosis bovina caudada por Campylobacter fetus produce pérdidas reproductivas existiendo poca información de los mecanismos de patogenicidad de dicha bacteria, por lo cual se propone un modelo utilizando ratones BALC/c. Embriones de dos células fueron cultivados en Ham-F10: grupo control (GC), los grupos experimentales fueron adicionados con las cepas Cfv 1, Cfv 3, Cff 2 y Cff 5. Mórulas fueron cultivadas en Ham-F10 (GC); los grupos tratados recibieron Cfv27, CFF (filtrado libre de células) y caldo Brucella (BB). Blastocistos fueron cultivados en MEM (GC) y MEM más Cfv 27 (grupo desafiado). Se evaluó: diferenciación, "hatching", "hatched", adhesión y expansión. Los resultados fueron analizados por Chi2. En embriones de dos células, la diferenciación no fue modificada por acción de las cepas evaluadas (p > 0,05). Para embriones en estadío de mórula, la diferenciación a las 24 h de cultivo fue menor para BB, Cfv, y CFF, comparado con el GC (p < 0,05). Luego de 48 h de cultivo, no hubo diferencias entre Cfv, BB, y CG (p > 0,05), no obstante para el grupo CFF la diferenciación fue menor al CG (p < 0,05). El porcentaje de "hatching" (48 y 72 h de cultivo), fue mayor en los grupos CFF y Cfv comparado con el GC (p < 0,05). La adición de Cfv 27 no modificó el desarrollo de blastocistos. En el modelo propuesto, Cf no afectó negativamente el desarrollo embrionario. Futuros trabajos serán necesarios para establecer un modelo de infección in vitro en pos de reproducir su patogenicidad.

Carcinoma de endometrio: revisión clínico-patológica de 6 años en relación con los factores pronósticos / Endometrial carcinoma: Clinico-pathological revision after 6 years, related to prognostic factors

Los carcinomas de endometrio se dividen en dos categorías mayores (I y II), según los datos clínico-patológicos y las alteraciones genéticas. Los de tipo I se asocian con hiperestimulación estrogénica, obesidad, tratamiento hormonal exógeno y reconocen como lesión precursora a la hiperplasia endometrial con desarrollo de carcinomas endometrioides (CE). Los de tipo II predominan en mujeres postmenopáusicas, de subtipos histológicos más frecuentes serosos y de células claras. El objetivo es realizar una revisión anátomo-clínica de los carcinomas de endometrio, estudiados en nuestra institución entre el periodo comprendido entre 1/01/2005 y 31/12/2010. Se realizó estudio retrospectivo y descriptivo de 234 casos de pacientes con diagnóstico de carcinoma endometrial. La edad promedio de las pacientes fue de 68,6 años. El motivo de la consulta en la mayoría de los casos fue metrorragia de la postmenopausia (91,2 por ciento). El factor de riesgo más frecuente fue hipertensión arterial (22 pacientes). El 67,5 por ciento de las pacientes tenían antecedentes de biopsia previa por videohisteroscopía realizada en nuestra institución, observándose concordancia diagnóstica en un 91 por ciento. Se clasificaron según la OMS en CE (79 por ciento), adenocarcinoma seroso (9 por ciento) y mixtos (12 por ciento). El grado histológico (GH) FIGO fue en el 86 por ciento tipo I (exclusión de carcinomas serosos) y el estadío más frecuente fue el I ( 65 por ciento). La metaplasia (Me) más frecuente observada en estas neoplasias fue la mucinosa (35 por ciento). Las mórulas se presentaron en 2 casos comprobados por IHQ con positividad para CD 10 y CDX-2 y negatividad para p63. Se evidenciaron cambios reactivos en el 26 por ciento. El patrón MELF (glándulas microquísticas, elongadas y fragmentadas), de infiltración en la pared miometrial, se identificó en 3 casos. El CE fue el tipo histológico más frecuente y su presentación (mayoritariamente en estadios tempranos de enfermedad), se asoció en un alto porcentaje con las metaplasias mucinosa y tubaria...

Endometrial carcinomas are divided into two major categories (I and II), according to clinicopathologic and genetic alterations. The type I is associated with estrogen hyperstimulation, obesity, exogenous hormonal treatment, and endometrial hyperplasia is recognized as a precursor lesion, with the consecuent endometroid carcinoma. The type II predominate in post-menopausal women, most common histologic subtypes are serous and clear cell. The aim is to review the anatomical and clinical characteristic of endometrial carcinomas, studied at our institution between 01/01/2005 and 31/12/2010. We performed a retrospective and descriptive study of 34 cases of patients with endometrial carcinoma. The average age was 68.6 years. The reason for consultation in most cases was post-menopausal's metrorrhagia (91.2 percent) Hypertension (22 patients) was the more prevalent risk fctor. 67.5 percent of the patients had a history of prior biopsy (histeroscopy) performed at our institution; diagnostic concordance was observed in 91 percent. Were classified according to the WHO in endometrioid carcinomas (79 percent), serous adenocarcinoma (9 percent) and mixed (12 percent). FIGO histologic grade was 86 percent in type I (excluding serous carcinomas) and stage I was the most frequent (65 percent). Mucinous metaplasia was the most frequently observed (35 percent). The morulae were presented in 2 cases (IHC positive for CDX-2 and CD10, and negative for p63). Reactive changes were in 26 percent. The MELF pattern of myometrial infiltration was identified in 3 cases. Endometrioid carcinoma was the most common histological type and presentation (mostly in early stages of disease). Was associated with a high percentaje of tubal and mucinous metaplasia (this would indicate malignancy attenuated). Not yet known meaning of morulae in carcinomas. While it must be record horns infiltration, the presence or not of the same does not change the treatment.

Carcinoma de endometrio: revisión clínico-patológica de 6 años en relación con los factores pronósticos / Endometrial carcinoma: Clinico-pathological revision after 6 years, related to prognostic factors

Los carcinomas de endometrio se dividen en dos categorías mayores (I y II), según los datos clínico-patológicos y las alteraciones genéticas. Los de tipo I se asocian con hiperestimulación estrogénica, obesidad, tratamiento hormonal exógeno y reconocen como lesión precursora a la hiperplasia endometrial con desarrollo de carcinomas endometrioides (CE). Los de tipo II predominan en mujeres postmenopáusicas, de subtipos histológicos más frecuentes serosos y de células claras. El objetivo es realizar una revisión anátomo-clínica de los carcinomas de endometrio, estudiados en nuestra institución entre el periodo comprendido entre 1/01/2005 y 31/12/2010. Se realizó estudio retrospectivo y descriptivo de 234 casos de pacientes con diagnóstico de carcinoma endometrial. La edad promedio de las pacientes fue de 68,6 años. El motivo de la consulta en la mayoría de los casos fue metrorragia de la postmenopausia (91,2 por ciento). El factor de riesgo más frecuente fue hipertensión arterial (22 pacientes). El 67,5 por ciento de las pacientes tenían antecedentes de biopsia previa por videohisteroscopía realizada en nuestra institución, observándose concordancia diagnóstica en un 91 por ciento. Se clasificaron según la OMS en CE (79 por ciento), adenocarcinoma seroso (9 por ciento) y mixtos (12 por ciento). El grado histológico (GH) FIGO fue en el 86 por ciento tipo I (exclusión de carcinomas serosos) y el estadío más frecuente fue el I ( 65 por ciento). La metaplasia (Me) más frecuente observada en estas neoplasias fue la mucinosa (35 por ciento). Las mórulas se presentaron en 2 casos comprobados por IHQ con positividad para CD 10 y CDX-2 y negatividad para p63. Se evidenciaron cambios reactivos en el 26 por ciento. El patrón MELF (glándulas microquísticas, elongadas y fragmentadas), de infiltración en la pared miometrial, se identificó en 3 casos. El CE fue el tipo histológico más frecuente y su presentación (mayoritariamente en estadios tempranos de enfermedad), se asoció en un alto porcentaje con las metaplasias mucinosa y tubaria...(AU)

Endometrial carcinomas are divided into two major categories (I and II), according to clinicopathologic and genetic alterations. The type I is associated with estrogen hyperstimulation, obesity, exogenous hormonal treatment, and endometrial hyperplasia is recognized as a precursor lesion, with the consecuent endometroid carcinoma. The type II predominate in post-menopausal women, most common histologic subtypes are serous and clear cell. The aim is to review the anatomical and clinical characteristic of endometrial carcinomas, studied at our institution between 01/01/2005 and 31/12/2010. We performed a retrospective and descriptive study of 34 cases of patients with endometrial carcinoma. The average age was 68.6 years. The reason for consultation in most cases was post-menopausals metrorrhagia (91.2 percent) Hypertension (22 patients) was the more prevalent risk fctor. 67.5 percent of the patients had a history of prior biopsy (histeroscopy) performed at our institution; diagnostic concordance was observed in 91 percent. Were classified according to the WHO in endometrioid carcinomas (79 percent), serous adenocarcinoma (9 percent) and mixed (12 percent). FIGO histologic grade was 86 percent in type I (excluding serous carcinomas) and stage I was the most frequent (65 percent). Mucinous metaplasia was the most frequently observed (35 percent). The morulae were presented in 2 cases (IHC positive for CDX-2 and CD10, and negative for p63). Reactive changes were in 26 percent. The MELF pattern of myometrial infiltration was identified in 3 cases. Endometrioid carcinoma was the most common histological type and presentation (mostly in early stages of disease). Was associated with a high percentaje of tubal and mucinous metaplasia (this would indicate malignancy attenuated). Not yet known meaning of morulae in carcinomas. While it must be record horns infiltration, the presence or not of the same does not change the treatment.(AU)