Preimplantation development of mouse and human embryos biopsied at cleavage stages using a modified displacement technique

A modified embryo biopsy method was tested on four- and eight-cell stage mouse embryos and used on human embryos to obtain blastomeres for preimplantation genetic diagnosis. The biopsy method tested combines zona drilling and fluid displacement to force one or two cells through an opening in the zona pellucida of the cleavage-stage embryo. Rates of cell division and the percentage of mouse embryos forming blastocysts following biopsy at the eight-cell stage were not significantly different from those observed in unoperated control embryos. The percentage blastocyst formation was not significantly different in embryos biopsied at the four-cell stage and in control embryos, although cell division was significantly retarded following biopsy. 96% of the mouse blastomeres isolated at the eight-cell stage were recovered intact and 96% of those placed in culture underwent cell division. Survival and division of cells isolated at the four-cell stage were 92 and 84% respectively. Most of the cultured blastomeres cleaved several times and formed small trophoblast vesicles. Chromosomes were observed in 59% of blastomeres incubated in the presence of colcemid. In the initial use of this biopsy technique for human preimplantation genetic diagnosis, blastocyst formation was observed in 9 of 13 human embryos biopsied at the 7- to 10-cell stage. These findings support the use of this biopsy method as an alternative to aspiration techniques.      

Cytogenetic analysis of spermatozoa in the father of a child with a de- novo reciprocal translocation t(7;9) (q22;p23)

Analysis of sperm chromosomes was carried out in the father of a child with a de-novo reciprocal translocation t(7;9) (q22;p23) by G-banding and chromosome painting. Sperm metaphases were obtained using the zona- free hamster oocyte-human sperm fusion technique. A total of 138 complements were sequentially analysed by G-banding and fluorescence in- situ hybridization (FISH). The frequency of spermatozoa with structural chromosome abnormalities (5.1%) and the estimated conservative aneuploidy (1.4%) were within the range obtained in our control donors (6.9 and 4%). The sex ratio (45.3% X versus 54.7% Y) was not significantly different from the theoretical 1:1. A total of 309 sperm complements was analysed by FISH, 138 sequentially analysed by G- banding-FISH and another 171 analysed by FISH only. The frequencies of structural chromosome abnormalities for chromosomes 7 and 9 (0.6 and 0% respectively) were not significantly different from those obtained in our control donors (0.6 and 0.8%). No spermatozoa with the t(7;9) (q22;p23) were observed, showing no evidence for a germ-cell mosaicism. A statistically significant, positive association between sperm breakpoints and fragile sites (P = 0.0225) was observed. However, the coincidence between fragile sites and sperm breaks (80%) was not significantly different from that obtained in our control donors (79.2%). These results suggest that in this case the risk of structural chromosome abnormalities in further offspring is not increased, although an association between fragile sites and sperm chromosome breaks in the father does exist.      

C-cells of the thyroid gland in experimental osteoporosis

The population of thyroid C-cells of female rabbits with ovariectomy-induced osteoporosis was studied using immunohistochemical method with antibodies against calcitonin (CT) and morphometrically. The development of osteoporosis was confirmed using radiology and densitometry. 8 weeks after the operation, after the detection of hypercalcemia and hypercalciuria, some of the animals were given a correcting treatment with calcium-containing drug and vitamin D3 or with a combination of calcium drug with hormonal estrogen-containing drug. 4 weeks after the treatment was started, thyroid C-cells in animals that received the drugs, were significantly larger than in the cells of control and ovariectomized rabbits that received no treatment. Following 12 and 22 weeks, in the animals treated with hormonal drug, both calcium concentrations in blood and urine and C-cell dimensions were normalized. Large C-cells with an intensive reaction to CT were observed at weeks 12 and 22 (weeks 20 and 30 after the operation) in the thyroid gland of the animals that received no treatment and of the animals that received calcium with food. In the last case, the morphological changes of C-cells were more pronounced, despite the insignificant differences in blood calcium concentrations in both groups. These results permit to suggest that the intensity of CT synthesis by C-cells may be influenced by calcium concentrations not only in the blood, but also in the intestinal lumen.     

Changes in binding of calcium ions and in function of Na+, K+-ATPase in the erythrocytes in essential hypertension

More calcium was removed by EDTA from the membranes of erythrocytes of patients with hypertension than from the erythrocytes of persons with a normal blood pressure. By the use of an isotonic solution of MgCl₂, the quantity of calcium removed from the erythrocyte membrane was the same in both groups. Activity of Na, K-ATPase in the membrane of reconstituted erythrocytes of patients with essential hypertension increased its activity in healthy human erythrocytes.No. 4 Main Board, Ministry of Health of the USSR. (Presented by Academician of the Academy of Medical Sciences of the USSR E. I. Chazov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 7, pp. 41–44, July, 1977.     

Acute cerebrovascular accidents with severe ovarian hyperstimulation syndrome

Cerebrovascular complications are by far the most serious side-effects of ovarian hyperstimulation syndrome. We report a case in which the patient developed cerebral infarction with right sided hemiplegia as a result of severe hyperstimulation syndrome after using a gonadotrophin- releasing hormone agonist for intracytoplasmic sperm injection.