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[Pregnancy and embryotoxicity of Rhizoma pinelliae in rats].

This experiment was conducted with sexually mature, virgin female rats. The body weight of the female rats was 230-350 g, that of male rats was 300-400 g. The female and male rats were mated overnight (the ratio was 3:1), the pregnant rats were divided into 7 groups. All drugs were given p.o. from the 6th to the 15th days of gestation. Pregnant rats were weighted on the 0th, 6th, 10th, 13th and 16th days of gestation. Vaginal bleeding of pregnant rats was examined on the 8th, 10th, 12th, 14th and 16th days of gestation. The rats were killed on the 20th day of gestation so as to count the number of implantation, dead and living fetuses. The living fetuses of rats were removes, weighted and inspected for gross abnormalities; the internal and skeletal malformations of live fetuses were examined. The following results were obtained: 9 g/kg dose of the crude Rhizoma Pinelliae powder was found to have the action in increasing the number of pregnant individuals with vaginal bleeding and dead embryo significantly. Comparing with the control, 9 g/kg dose of the powder of processed Rhizoma Pinelliae did not show any toxicity. But 30 g/kg dose of the decoction of processed or crude Rhizoma Pinelliae was found to increase the number of pregnant individuals with vaginal bleeding and dead embryo significantly.

Abnormalities, Drug-Induced↗

Prevention of tilorone developmental toxicity with progesterone.

The immunomodulator tilorone hydrochloride was administered (gastric intubation) once to time-pregnant Upj:TUC(SD)spf (Sprague-Dawley) rats in four experiments. In experiment 1, tilorone (250 or 500 mg/kg) was administered on day 10 of gestation. The dams were killed 4 or 72 hr after dosing. Interferon-like activity and drug levels were determined in maternal blood, spleen, and thymus, as well as in the embryos. In experiment 2, the test groups received progesterone (2 mg/kg), or tilorone (200 or 400 mg/kg), or progesterone and tilorone. The dams from each group were killed 24 or 48 hr after receiving tilorone. Experiment 3 was similar to experiment 2, except that the dams were killed on gestation day 20. In experiment 4, tilorone (400 mg/kg) was administered on gestation day 17, 18, or 19, and the dams were killed 24 hr after dosing or on gestation day 20. In all four experiments, tilorone-related maternal toxicity (regardless of whether progesterone also was administered) was observed, as characterized by marked decreases in weight gain, the occurrence of clinical signs, and in experiment 1 by decreased thymus weights, 72 hr post-dosing. Dose-related increases in the mean number of dead embryos and in serum interferon titers occurred 72 hr postdosing. In experiment 2, there was an increase in the number of dams in the 400-mg/kg (tilorone only) group with dead embryos only, 24 hr postdosing; similar results occurred in both the 200- and 400-mg/kg groups, 48 hr postdosing. However, in the groups that also received progesterone, a partial prevention of such embryolethality was evident. In experiment 3, embryotoxicity again was observed in both tilorone-treated groups, whereas several of the dams that were also given progesterone through day 19 of gestation experienced at least a partial prevention of the embryolethal effects of tilorone. In experiment 4, no fetotoxicity was observed despite the severe maternal toxicity evident.

Animals↗

Teratologic potential of 2-methoxyethanol and transplacental distribution of its metabolite, 2-methoxyacetic acid, in non-human primates.

The embryotoxic effects of 2-methoxyethanol (2-ME) were studied in non-human primates to better assess the risk for women of child-bearing age exposed to this agent. Macaca fascicularis females were treated daily throughout the organogenetic phase of pregnancy (days 20-45) by gavage and the fetuses collected at day 100 by Caesarean section. At the highest dose (0.47 mmole/kg), all eight pregnancies ended in death of the embryo. One of these dead embryos was abnormal, missing a digit on each forelimb. At the middle dose (0.32 mmole/kg), three of 10 pregnancies ended in embryonic death, presumably due to 2-ME exposure and three of 13 pregnancies met a similar fate at the low dose (0.16 mmole/kg). In each of these two groups, an additional pregnancy was lost to abortion, but both were thought to be spontaneous, which usually occurs in 10-20% of untreated macaque pregnancies. These results indicate that 2-ME is a potent toxin to the developing primate embryo and thereby furthers the concern about exposure of pregnant women to this agent, although maternal toxicity was evident in nearly all treated pregnancies and was especially severe in the high-dosage animals. Distribution of the major metabolite of 2-ME, 2-methoxyacetic acid (2-MAA), indicated a long half-life (ca. 20 h), resulting in accumulation of metabolite in maternal serum after repeated daily dosing. Transplacental studies revealed uniform distribution in the embryo and extraembryonic fluids at a concentration similar to that in maternal serum. The yolk sac, on the other hand, accumulated a very high concentration of 2-MAA, but the embryotoxic significance of this observation is unknown.

Acetates↗

Transplacental infection following exposure of gilts to porcine reproductive and respiratory syndrome virus at the onset of gestation.

Twenty-five gilts without measurable porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) serum antibody titres were used for this experiment. All of them were randomly assigned to one of the treatment groups at the time of artificial insemination. Twelve gilts were exposed to PRRSV, of these, six were slaughtered on day 10 after exposure and constituted group A. The remaining six were slaughtered on day 20 after infection and constituted group C. Thirteen gilts were used as controls, six of these were slaughtered on day 10 after treatment and constituted group B. The remaining seven were slaughtered on day 20 after treatment and constituted group D. The infected gilts were inoculated with PRRSV intranasally and intravenously in the ear vein. They were observed for clinical signs of infection and the effects on conception and fertilization rates were studied, while the gilts and their embryos were tested for PRRSV and homologous antibodies. The infected animals developed signs of PRRS associated with anorexia and slight pyrexia. Infection was verified by reisolation of the virus from serum and other tissue samples and also by seroconversion. Ten out of 12 infected gilts and 10 out of 13 controls were pregnant at the time of slaughter and the ratio of embryos to corpora lutea was the same in both, infected and control groups (0.75). Therefore, infection with PRRSV at the onset of gestation did not appear to interfere with conception and fertilization rates and subsequent pregnancy. The PRRSV was not isolated from any of the embryos collected at day 10 postexposure, but was present in 20-day-old embryos of group C gilts. In this group, 60% of litters were infected prenatally, with 16% of embryos infected. The proportion of dead embryos was three times greater than in a control group D (35.4% and 9.8%, respectively). The results of this report indicate that exposure of susceptible gilts to PRRSV at the onset of gestation has no significant effect on conception and fertilization rates. However, although infection does not appear to have any effect on the embryos before implantation, it can result in transplacental infection and embryo death.

Animals↗

Do pregnant lizards resorb or abort inviable eggs and embryos? Morphological evidence from an Australian skink, Pseudemoia pagenstecheri.

Although pregnant viviparous squamates are sometimes claimed to be able to resorb inviable eggs and embryos from the uterus, definitive evidence for such resorption is not available. After placing pregnant female Pseudemoia pagenstecheri into conditions under which embryonic development is terminated, we periodically harvested the gravid oviducts and examined them histologically. Females contained abnormal and degenerating eggs and embryos that had died in various stages of development. Dead embryos had undergone extensive cytolysis, dissolution, and aseptic necrosis and vitelline masses showed signs of deterioration and passage down the oviduct. The uterine mucosa lay in direct contact with the vitelline material, with no intact shell membrane intervening between them. Yolk was sometimes displaced into the exocoelom and allantoic cavity due to rupture of the extraembryonic membranes. Histological examination revealed no evidence of the uptake of yolk by the uterine epithelium or its accumulation in the subepithelial connective tissue. In many specimens, the uterine epithelium showed minuscule, apical granules. The position, appearance, and staining properties of the granules suggests them to be secretory, a manifestation of placentotrophy. Our observations indicate that P. pagenstecheri females retain dead eggs and embryos for several weeks or longer, yet do not resorb them during that period. This lizard is the second placentotrophic skink species in which resorption has been suspected, but in which abortive eggs appear to be retained or extruded instead of being resorbed by the oviducts. Researchers should not assume that squamates can digest and resorb oviductal eggs without definitive morphological evidence.

Animals↗

Pathological and immunological study of goose embryos and goslings inoculated with an attenuated strain of Derzsy's disease virus.

An attenuated Derzsy's disease virus strain, designated BAV, was studied in goose embryos. A total of 248 embryonated goose eggs, coming from a susceptible laying flock with no yolk-derived immunity (group I) and from a vaccinated laying flock (group II) were used. The eggs were inoculated into the allantoic cavity with 10(1.9), 10(2.9) or 10(3.9) EID 50/0.2 ml virus on day 12 or day 20 of incubation. Embryos were killed at 5-day intervals. The dead embryos and the hatched goslings (up to 2 weeks of age) were examined by gross and histopathological methods. Reisolation of the virus from the organs was attempted, serum samples were tested for the presence of antibodies, and lymphocytes separated from the circulating blood were used in the lymphocyte stimulation and immunorosette formation tests. Embryos of both groups I and II, inoculated at either time of incubation, showed a body mass gain inferior to that of the controls. Sixteen (group I) and 12 (group II) of the embryos inoculated on day 12 of incubation died. Some (group I: 15, group II: 6) of the embryos inoculated on day 20 of incubation failed to hatch. The pathomorphological changes seen in the embryos killed between days 17 and 22 of incubation were of degenerative character. In embryos killed later (between days 23 and 58 of incubation) the degenerative changes were accompanied by infiltration by inflammatory cells. Reisolation of the virus strain was mostly successful between postinoculation (PI) days 5 and 10. Specific virus-neutralizing antibodies and cellular immune response were demonstrable already at hatching.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Development of preimplantation mouse embryos after exposure to a 50 Hz magnetic field in vitro.

Effect of sinusoidal 50 Hz magnetic field (MF) on development of preimplantation CBA/S mouse embryos in vitro was studied. Superovulated and in vivo fertilized preimplantation embryos were collected at one cell stage and divided to control and MF-exposed groups. Sinusoidal 50 Hz MF with field strength of 10 A/m r.m.s., corresponding a flux density of 13 microT r.m.s., was used to expose the embryos in culture at 37 degrees C in a CO2-incubator. The developmental stage and abnormalities were recorded twice daily except once daily during weekends. The vitality and developmental stages of the embryos were similar in both groups although slightly more dead embryos were found during the 1st day in MF-exposed group (P<0.05) and the development of MF-exposed embryos was slightly impaired. In conclusion, the exposure to sinusoidal 50 Hz MF at field strength of 10 A/m did not significantly disturb the development of the mouse embryos in vitro up to the blastocyst stage.

Animals↗

[Effect of being fed gamma-irradiated fish on embryogenesis and the chromosomes of white rats].

Experiments on Wistar rats were made to study potential embryotoxic and cytogenetic effects of fresh and smoked fish subjected to gamma-irradiation. Embryotoxic action of irradiated fish was studied in 611 embryos obtained from 52 females. The number of alive and dead embryos, their mass, length and the presence of external and internal developmental abnormalities were determined. the animals' bone marrow cells were subjected to cytogenetic examination with the use of the metaphasic method. A total of 2426 cells from 35 rats were examined for the occurrence of chromosome aberrations. The examination has revealed no embryotoxic or cytogenetic action of both fresh and smoked fish subjected to gamma-irradiation.

Animals↗

Aerobic bacterial flora from dead-in-shell chicken embryos from Nigeria.

Dead-in-shell chicken embryos from two commercial hatcheries in Anambra State of Nigeria were investigated for isolation of aerobic bacteria. For this purpose, 79 pooled samples containing 632 dead-in-shell chicken embryos were cultured. From these samples, 23 isolates of Escherichia coli and 25 isolates of Staphylococcus aureus were recorded. Other bacterial species isolated included Micrococcus sp. (fifteen isolates), Klebsiella sp. (thirteen isolates), Pseudomonas sp. (nine isolates), and Proteus sp. (seven isolates). Salmonella, Streptococcus, and Mycoplasma spp. could not be isolated. A high incidence of pathogenic strains of bacteria from dead-in-shell chicken embryos was observed. This suggests that the isolates may have contributed to the embryonic mortality and reduced hatchability recorded in the farms investigated.

Animals↗

Abnormal development and transport and increased sister-chromatid exchange in preimplantation embryos following superovulation in mice.

Both sister-chromatid exchange (SCE) response and embryonic development and transport in preimplantation embryos were evaluated on day 3 of gestation (vaginal plug = 1) of superovulated Swiss mice. Superovulation was found to have significant effects on number of preimplantation embryos (increase), embryo localization (accelerated transport), cleavage rate (advanced development) and abnormality rate (misshaped, fragmented, dead embryos). Superovulated 4- and 8-cell embryos collected from oviducts and uteri and incubated in vitro with 5-bromodeoxyuridine (BrdU) displayed up to 4 times higher SCE frequency than spontaneously ovulated embryos. This increase is independent of stage of development and location at the time of embryo collection. The results indicate that superovulated embryos may have induced DNA lesions.

Abnormalities, Drug-Induced↗

Vitamin E in early stages of sea bass (Dicentrarchus labrax) development.

This study reports titration of vitamin E levels in the sea bass (Dicentrarchus labrax) using high-pressure liquid chromatography. The first part of the work is devoted to vitamin E detection in: (1) plasma of maturing females and males characterized by different body sizes; (2) seminal fluid and eggs; and (3) developing embryos of sea bass fed with vitamin E. In the second part of the study, variations of vitamin E levels during larval development are analyzed. The results show a direct correlation between plasma vitamin E content and body size for both adult male and female sea bass. High vitamin E levels were found in seminal fluid, in eggs before and after fertilization, and in embryos during development and at hatching, whereas vitamin E level was low in dead embryos and in embryos with limited survival. During larval development, the vitamin E content decreased slowly but steadily during the first four days of larval growth; subsequently, it progressively increased from day 9 to day 40. In teratogenic larvae, vitamin E content was significantly higher than in normal larvae. This study provides evidence on how vitamin E exerts an antioxidant defense in sea bass reproduction.

Animals↗

Identification and isolation of embryonic stem cells in reproductive endocrinology: theoretical protocols for conservation of human embryos derived from in vitro fertilization.

BACKGROUND: Embryonic stem cells (ESC) are pluripotent cells obtained from the inner cell mass (ICM) of blastocysts derived from in vitro culture associated with reproductive endocrinology therapy. Human ESCs are regarded as highly significant since they retain the capacity to differentiate into any of approximately 200 unique cell types. Human ESC research is controversial because to acquire such cells, the ICM of human blastocysts must be manipulated in a way that renders embryos nonviable and unsuitable for transfer in utero. Techniques to yield competent ESCs with conservation of source blastocysts would satisfy many objections against ESC research, but at present such approaches remain largely untested. RESULTS AND DISCUSSION: We contrast experimental culture of single blastomeres obtained by 1) non-destructive biopsy of embryos destined for transfer, and 2) isolation of karyotypically normal blastomeres from disaggregated ("dead") embryos considered unsuitable for transfer, and evaluate these approaches with regard to production of ESCs. Pluripotency was confirmed by morphological criteria and by quantification of divergent homeodomain proteins specific to undifferentiated cell development. Following ESC isolation and identification, assessment was conducted according to a novel ESC grading system, also proposed here. CONCLUSION: The role of reproductive endocrinology in ESC research remains paramount. In this report, we hypothesize new and expand on existing strategies having the potential to enhance human ESC isolation, identification and in vitro maintenance.

Animals↗