The inheritance, replication and perpetuation of the sperm centriole in the early human embryo are reported. Both normal monospermic and abnormal dispermic embryos (n = 127) were examined by transmission electron microscopy. Centrioles were traced from fertilization to the hatching blastocyst stage. The sperm proximal centriole is introduced into the oocyte at fertilization and remains attached to the expanding spermhead during sperm nuclear decondensation, as it forms the male pronucleus. A sperm aster is initially formed after the centriole duplicates at the pronuclear stage. At syngamy, centrioles occupy a pivotal position on opposite spindle poles, when the first mitotic figure is formed. Bipolar spindles were found in the majority of embryos, while tripolar spindles were seen in four dispermic embryos at syngamy. Two single centrioles were detected at two poles of two tripolar spindles, while two additional centrioles were located on the sides of a bipolar spindle of a dispermic embryo. Sperm tails were detected near spindle poles at syngamy and in later embryos. Typical centrioles showing the characteristic pin-wheel organization of nine triplets of microtubules were evident. During centriolar replication, the daughter centriole grows laterally from the parent and gradually acquires pericentriolar material (PCM). The two centrioles are surrounded by a halo of electron-dense PCM, which nucleates microtubules, thus making it a typical centrosome. The usual alignment of diplosomes at right angles to each other was maintained. Centrioles were detected at all stages of embryonic cleavage from the 1-cell through 8-cell stages, right up to the hatching blastocyst stage. They were closely associated with nuclei at interphase, when they were often replicating, and were prominently located at spindle poles during the first four cell cycles. In blastocysts, they were detected in trophoblast, embryoblast and endoderm cells respectively. It is evident that the sperm centrosome is the functional active centrosome in human, while the female is inactive but may contribute some centrosomal material to the zygote centrosome. It is very likely that the paternal centriole is the ancestor of the centrioles in fetal and adult somatic cells.

Totipotent non-committed inner cell mass (ICM) cells from human blastocyts, if demonstrated to be capable of proliferating in vitro without differentiation, will have several beneficial uses, not only in the treatment of neurodegenerative and genetic disorders, but also as a model in studying the events involved in embryogenesis and genomic manipulation. Nine patients admitted to an in-vitro fertilization programme donated 21 spare embryos for this study. All 21 embryos were grown from the 2-pronuclear until blastocyst stages on a human tubal epithelial monolayer in commercial Earle's medium (Medicult, Denmark) supplemented with 10% human serum. The medium was changed after blastocyst formation to Chang's medium supplemented with 1000 units/ml of human leukaemia inhibitory factor (HLIF) and the embryos left undisturbed for 72 h to allow the hatched ICM and trophoblast to attach to the feeder monolayer. Nineteen of the 21 embryos from nine patients produced healthy ICM lumps which could be separated and grown in vitro. Two of the lumps differentiated into fibroblasts while the remaining 17 (eight patients) produced cells with typical stem cell-like morphology, were alkaline phosphatase positive and could be maintained for two passages. It was possible to retain the stem cell-like morphology, alkaline phosphatase positiveness and normal karyotype through the two passages in all of them using repeated doses of HLIF every 48 to 72 h. This is the first report on the successful isolation of human ICM cells and their continued culture for at least two passages in vitro.

A prospective study was conducted to compare the accuracy of visual estimation of blood loss (EBL) at delivery with laboratory determination of measured blood loss (MBL). It showed that EBL tends to be clouded by the conventional teaching that blood loss at delivery is usually between 200 to 300 mL. Women with MBL up to 150 mL were overestimated and the best correlation was in women with MBL between 150 to 300 mL. There was a tendency to underestimate blood loss when the MBL was between 301 to 500 mL. Of the 9 women with a primary postpartum haemorrhage, only one was correctly diagnosed as such and 3 women were estimated to have blood losses of at least 500 mL but the measured blood losses were all lower. It was concluded that visual estimation of blood loss is inaccurate, especially at the extremes of MBL and that primary postpartum haemorrhage is not detected by visual estimation of blood loss, unless there are associated signs of haemodynamic instability.

We demonstrate the presence of centrioles in fertilized human oocytes at syngamy. Single or double centrioles within centrosomes were detected by transmission electron microscopy at one pole of the first cleavage spindle in normal and dispernic embryos (25-26 hr after insemination). Sperm centrioles were also closely associated with the male pronucleus (16-20 hr after insemination) in pronuclear stage embryos. A tripolar spindle derived from a tripronuclear embryo is also demonstrated with two centrioles at one pole. The data provide evidence that human centrioles, as those in most other animals, and unlike the mouse, are paternally derived, thus supporting Boveri's dassical theory. Furthermore, this study provides insihts to the proposed mechanisms of aberrant cleavage patterns of dispermic human embryos.It is widely believed that mature mammalian oocytes and early cleavage stage embryos do not have centrioles (1-6). Most cells, however, do possess centrosomes, which are microtubule (MT) organizing centers at spindle poles (3). In his classical theory of fertilization, Boveri in 1900 (7) stated that unfertilized eggs derive their centrosomes from male gametes, and this has subsequently been shown to be the case in a number of animal species, including the sea urchin (2), where centrioles associated with centrosomes organize mitotic bipolar spindles (3). On the contrary, in mice, centrosomes are maternally derived (2) and this has been proposed to be true for other mammals.Meiotic spindles of mammalian oocytes are anastral, barrel shaped, and composed of numerous MTs (1, 2, 4). The structure of the human meiotic spindle has already been described to conform to the mammalian pattern (5,6,8,9). Mammalian meiotic spindles have centrosomes but no centrioles. Centrosomes and centrioles are both self-reproducing organelles and centrioles merely advertise the presence of centrosomes (3). After fertilization, the mitotic spindle of the sea urchin embryo is organized by paternally inherited centrioles and centrosomes (10, 11). In the sea urchin, each sperm carries two centrioles associated with centrosomes (12), which duplicate and separate to form a bipolar spindle during the first mitosis and are the ancestors of these organelles in all cells during subsequent development (2, 10, 11). In a fashion similar to sea urchin sperm, human sperm also have centrioles. A well-defined proximal centriole is present next to the basal plate of the sperm head (13-15), while the distal centriole (which is a remnant) gives rise to the sperm tail axoneme during spermiogenesis. The proximal centriole consists of nine triplets of MTs showing the typical 9 + 0 organization and is associated with osmiophilic centrosomal material. After gamete fusion, the sperm midpiece and tail are invariably incorporated into the ooplasm, and the centriolar region often remains attached to the decondensing sperm nucleus and persists after male pronuclear formation (16-18). This study demonstrates the presence of centrioles associated with centroso...

Placenta accreta is a rare condition and is associated with considerable maternal morbidity and mortality. Though the surgical approach of hysterectomy is a definitive therapy, there are occasions when conservation of the uterus is desired by the patient. We report a case of placenta accreta successfully treated with intravenous methotrexate. After 2 weeks of treatment no signs of placenta could be visualized on ultrasound examination of the uterus. The patient was discharged after 15 days and has since been well. Such therapy may be useful in exceptional cases in institutions with adequate facilities for careful monitoring and management of the patient.

No abstract

Cultured human, ampullary, epithelial cells obtained from fertile women undergoing hysterectomy were evaluated for the support of human embryonic cleavage and growth in vitro. Twelve patients provided 23 embryos for co-culture with subcultured ampullary cells grown in T6 + 15% patient's serum and 18 embryos for growth in T6 + 15% patient's serum alone (controls). Of embryos co-cultured with ampullary cells, 78% cleaved to the compacted embryo stage and 69% cavitated as compared with 50 and 33% respectively for controls (P less than 0.01). Only 30% of co-cultured embryos reached the expanded blastocyst and 26% underwent hatching as compared with 28% for both stages in controls. At the 2 - 4- and 6 - 8-cell stages, 91 and 87% of co-cultured embryos showed an absence or slight fragmentation as compared with 72 and 61% respectively for embryos grown in medium alone (P less than 0.01). None of the co-cultured embryos showed unequal-sized blastomeres while 22% of controls showed unequal cleavage. Embryos grown with ampullary cells cleaved slightly faster than controls. Scanning electron micrographs showed that ampullary cells collected from co-cultures were all of the secretory type with several microvilli and apical protrusions. It is clear that subcultured human ampullary cells support human embryonic cleavage and yield a reasonable number of good quality embryos up to the cavitation stage. Development past the expanded blastocyst and hatching stages seems to involve another critical phase with its own specific requirements.

During a period of 8 years (1985-92), 100 fetuses were diagnosed to have non-immune hydrops on the basis of ultrasonographic findings and absence of rhesus isoimmunization. Both the mother and the fetus were thoroughly evaluated by a set protocol that included a detailed fetal abnormality scan with echocardiography and fetal blood sampling. A cause for non-immune hydrops could be identified in 81% of the fetuses. Cardiovascular abnormalities (23%) and alpha(1)-thalassemia (22%) were almost equally common etiological factors in the South-East Asian population under investigation. A chromosomal abnormality was detected in 10% of the fetuses with non-immune hydrops. Twenty-six fetuses were found to be suitable for in utero therapy. In utero therapy included one or more of the following: (1) fetal intravascular blood transfusion; (2) direct fetal drug therapy; and (3) fetal pleuroamniotic shunting. Eighteen of the 26 babies (69.2%) were alive and well at 1 month after delivery. It is concluded that in well-selected cases appropriate in utero fetal therapy can lead to significant improvement in fetal salvage.