Histologically, immunohistochemically, ultrastructurally, and molecularly confirmed neosporosis abortion in an aborted equine fetus.

Neosporosis is a common cause of abortion in cattle worldwide but is rare in horses. Here, the first case of histologically, ultrastructurally, immunohistochemically, and molecularly confirmed equine abortion caused by neosporosis is reported. Samples of lung, heart, liver, skeletal muscle, tongue, brain, and the placenta from a female fetus aborted at 280 days of gestation were fixed in formalin and submitted for diagnosis. Histologically, there was disseminated neosporosis with severe lesions in lungs, liver and the heart. Protozoal tachyzoites in all tissues reacted with polyclonal anti-Neospora caninum rabbit antibodies. Transmission electron microscopic observation on lung tissue revealed tachyzoites consistent with Neospora, including many rhoptries. Polymerase-chain reaction (PCR) using primers designed to amplify the rRNA gene internal transcribed spacer 1 (ITS1) of the Sarcocystidae was performed on DNA extracted from fetal tissues. Comparison of the ITS1 amplified from the foal tissue to sequences available in GenBank revealed 100% sequence identity to the ITS1 from three isolates of Neospora hughesi.

Telomere length dynamics in early life: the blood-and-muscle model.

Telomere length (TL) trajectories in somatic tissues during human growth and development are poorly understood. We examined a blood-and-muscle model during early life, focusing on TL trajectories in leukocytes, representing the highly proliferative hematopoietic system, and skeletal muscle, a minimally proliferative tissue. Leukocyte TL (LTL) and skeletal muscle TL (MTL) were measured in 28 fetuses and 73 children. LTL and MTL were highly variable across individuals (sd: fetal LTL = 0.72 kb, MTL = 0.72 kb; children LTL = 0.81 kb, MTL = 0.82 kb) but were highly correlated within individuals (fetuses, r = 0.76, P < 0.0001; children, r = 0.87, P < 0.0001). LTL was shorter than MTL in fetuses (10.63 vs. 11.01 kb; P = 0.0004) and children (8.46 vs. 9.40 kb; <0.0001). The LTL-MTL gap was smaller in fetuses than children. TL in children was inversely correlated with body mass index (BMI) (LTL: -0.047 ± 0.016 kb/BMI, P < 0.005; MTL: -0.037 ± 0.017 kb/BMI, P = 0.03). We conclude that variations in TL across adults and differences in TL between somatic tissues are largely established in early life. Because TL plays a significant role in aging-related diseases, insight into the factors that fashion TL in somatic tissues during early development should contribute to an understanding of the relationship of TL with these disease and longevity in humans.-Sabharwal, S., Verhulst, S., Guirguis, G., Kark, J. D., Labat, C., Roche, N. E., Martimucci, K., Patel, K., Heller, D. S., Kimura, M., Chuang, D., Chuang, A., Benetos, A., Aviv, A. Telomere length dynamics in early life: the blood-and-muscle model.

Array comparative genomic hybridization and flow cytometry analysis of spontaneous abortions and mors in utero samples.

BACKGROUND: It is estimated that 10-15% of all clinically recognised pregnancies result in a spontaneous abortion or miscarriage. Previous studies have indicated that in up to 50% of first trimester miscarriages, chromosomal abnormalities can be identified. For several decades chromosome analysis has been the golden standard to detect these genomic imbalances. A major drawback of this method is the requirement of short term cultures of fetal cells. In this study we evaluated the combined use of array CGH and flow cytometry (FCM), for detection of chromosomal abnormalities, as an alternative for karyotyping. METHODS: In total 100 spontaneous abortions and mors in utero samples were investigated by karyotyping and array CGH in combination with FCM in order to compare the results for both methods. RESULTS: Chromosome analysis revealed 17 abnormal karyotypes whereas array CGH in combination with FCM identified 26 aberrations due to the increased test success rate. Karyotyping was unsuccessful in 28% of cases as compared to only two out of hundred samples with inconclusive results for combined array CGH and FCM analysis. CONCLUSION: This study convincingly shows that array CGH analysis for detection of numerical and segmental imbalances in combination with flow cytometry for detection of ploidy status has a significant higher detection rate for chromosomal abnormalities as compared to karyotyping of miscarriages samples.

[Molecular-cytogenetic study of the aborted fetuses in women with reproductive function disorders].

It is known that the frequency of chromosomal abnormalities among spontaneous miscarriages of the first trimester of pregnancy makes 50-60%. Research of karyotypes of chorionic villus cells of miscarriages has been conducted by combining the standard cytogenetic method and the FISH analysis on interphase nuclei of centromeric specific DNA samples by the tests to the chromosomes 13/21, 14/22, 15, 16, 18, X, Y. The described complex approach can be successfully applied for effective identification ofchromosomal abnormalities in the material of spontaneous miscarriages. The results specify the necessity of careful study of genomes of matrimonial pairs with the usual unmaturing in anamnesis and especially before treatment by IVF methods.

Morphological and ultrastructural evaluation of cultured frozen-thawed human fetal ovarian tissue.

OBJECTIVE: To investigate a defined culture condition for the culture of frozen-thawed human ovarian tissue. DESIGN: Prospective laboratory study. SETTING: Reproductive biology laboratories in university hospitals. PATIENT(S): Fetal ovarian tissue from elective termination of pregnancy. INTERVENTION(S): Culture of frozen-thawed fetal ovarian tissue for up to 63 days. MAIN OUTCOME MEASURE(S): Morphology, morphometry, and survival of follicles in relation to culture times. RESULT(S): The proportion of primordial, early primary, and primary follicles in frozen-thawed (day 0) ovarian tissue was 77.5%, 21.7%, and 0.8%, respectively. Pronounced degeneration was found in all cell types, and < or =36% of the follicles had signs of atresia at days 7-14, but this figure improved with culture time to <20% of the total follicular population. After 7-14 and 21-35 days of culture, the relative proportion of the follicles in the different classes remained nearly stable. Morphometric examination of healthy follicles showed a significant increase in both follicle and oocyte diameter compared with control. A few follicles had developed to the early secondary stage. Ultrastructural analysis demonstrated well-preserved morphological integrity of healthy primordial and early primary follicles. Immunohistochemical localization of proliferating cell nuclear antigen was positive in proliferating follicular cells at days 7-14 and 21-35 of culture. CONCLUSION(S): The present culture condition leads to good survival and progressive follicular growth and differentiation that is comparable to the physiological pattern of early folliculogenesis.

Emergence of a glomus-like body in the human yolk sac: a microanatomical analysis of its structure.

Since the last decade the Yolk sac (YS) has been a topic of increasing interest due to the growing use of high-resolution sonography in early determination of pregnancy. Human YS shape and diameter are indicators of viability of pregnancy during the early embryonic period. Nevertheless, the major interest concerns the vital function it plays in early embryo growth and development. Two compartments are recognized in this organ: the yolk sac proper and the vitelline stalk. In this study we report the identification and partial characterization of a glomus-like body in the wall of the secondary YS in humans. A detailed structural description is also presented on the time course of formation of this new structure, at precisely sequential stages between 4-8 wk post-conception. The significance of this new compartment on the YS function is analyzed. Light and scanning electron microscopy were used to investigate the microstructure of the YS and the vitelline stalk during the first 8 wk of development. Ten YSs were collected from embryos (aged between 24-50 days) obtained from emergency salpingectomies due to tubal ectopic pregnancy. From 5 wk onward a new structure was observed in the YS located near the apex of the pear-shaped yolk vesicle and closed to the connecting stalk. We designate this differentiation as glomus-like body. This structure is 1-1.5 mm long and merged from a pocket-like structure of the extraembryonic splanchnic mesoderm of the YS wall. It likely represents an area of convergence of the vascular network of the YS wall. Our findings underline the remarkable complexity of the human secondary yolk sac during early development. The detailed description of the microanatomy of this vital organ is of theoretical and practical interest in order to unravel the mechanisms used by the yolk sac to transport nutrients to the embryo.

Microscopical survey of the development and differentiation of the epithelium of the uterine tube and uterus in the human fetus.

The development and differentiation of the coelomic epithelium lining the paramesonephric ducts in human fetus, that gives rise to the female genital organs, have been ultrastructurally examined. The epithelium appeared pseudostratified, consisting of basal, microvillous and ciliated cells. In younger fetuses (12th gestational week) ciliogenic elements could be detected mainly on the developing tubal fimbriae, but most of the cells showed microvilli and often single cilia. In the subsequent phases of development, morphodynamics of cell renewal were documented by aspects of apoptosis. Fully ciliated cells were numerous on the fimbriae and at the utero-tubal junction, but not in the uterus; however, these were less abundant than those showing microvillous. In older fetuses (31st gestational week) microapocrine secretion by microvillous cells, in the form of droplets contacting cilia, could be observed. In the same fetuses the ectocervix was covered by a mature squamous epithelium, made up of polygonal flat desquamating cells, showing labyrinthine surface microplicae. Our observations demonstrated that ciliation in the human female genital organs, like that of other systems, is neither simultaneous nor uniform, and ciliated cells are gathered preferentially in strategic sites, to mediate germ cell migration and blastocyst implantation in adult life. These ultrastructural data seem to indicate that the female genital tract epithelium, at least in its general features, is sketched since fetal life, and cell morphodynamics, including microvillous and ciliated cell differentiation, as well as the secretory activity, are the morphological expression of the complex molecular mechanisms, involved in developmental biology and reproductive physiology.