Morphologic and histochemical characteristics of mammalian embryos during the pre-implantation period of development]

Submicroscopic embryo organization during preimplantation development was analyzed and its carbohydrate metabolism at these stages was studied. In the process of investigation the following regularities were revealed: progressive rearrangement of the nucleus (from compact to reticular); alterations in cytoplasmic structures (in multivesicular bodies, in autophagal vacuoles, in mitochondria, in granular cytoplasmic network, in lamellar structures); complication of intracellular interrelations (by means of microvilli and median bodies at early stages and during subsequent formation of cellular processes, contacts after "thorn--net" pattern, dense, fissure-like contacts and desmosomes). By means of cytochemical and electron histochemical methods, glycogen distribution in embryos at different stages of development was elucidated, and tendency to its decrease with the progress of the development was noted, with its least contents being at the stage of blastocyst.     

Biochemical characteristics of adhesion factors isolated from the retina of 8-day-old chick embryos

While isoelectrofocusing the extracts obtained after dissociation of the retina of eight day old chick embryos, two active protein fractions were found: AF I with pI 3 and AF II with pI below 3. The content of these fractions in the extracts depended on the method of dissociation: treatment with EDTA increased the content of AF II, whereas trypsin treatment increased the yield of AF I. After electrophoresis in the denaturating conditions both fractions were divided in two components. The effect of AF I on aggregation of retinal cells, unlike that of AF II, does not depend on the presence of bivalent cations in the medium and is realized within 1 h of interaction with the cells at 4 degrees. It is suggested that the retinal extracts of eight day old chick embryos contain substances providing two types of adhesiveness: Ca2+-independent (AF I) and Ca2+-dependent (AF II).     

Antitumor resistance induced by immunization with normal definitive tissues]

Immunization of C3HA mice with homogenates of various normal definitive tissues, obtained from syngeneic, allogeneic and xenogeneic donors, resulted in an increase of antitumor resistance, as evidenced by a considerable retardation of growth of the transplantable strain-specific hepatoma 22a. The effect may be due to sensibilization of animals to the antigens of normal definitive cells, and, in particular to the tissue-specific antigens characteristic of cells of the tumor in question.     

Adaptive characteristics of growth of human embryos and fetuses during the intrauterine life]

Under observations was the dynamics of growth of human embryos and fetuses in embryogenesis which is directly dependent on the month of the intrauterine life. Coefficients of correlation and regression were calculated and with their help the average growth of the general length of fetuses in each month of prenancy was determined. The enlargement of the fetus length was found to be more intensive in the embryonic and prefetal periods.     

Isolation and electrophoretic characteristics of 3 lymphocyte populations of normal human blood]

Cell electrophoresis allows separation of normal human blood lymphocytes into two main groups which are a function of their relative rates of migration, with regard to the reference speed (1 mum.sec.-1V-1.cm): the lymphocytes which have a greater mobility than this value seem to be T-lymphocytes (80,1 per cent for 42 healthy adults); on the contrary, B-lymphocytes have an inferior mobility (19,9 per cent). Two known methods are used for the selection of the lymphoid populations: spontaneous rosetting with sheep's red blood cells, which are characteristic of T lymphocytes, and adherence to nylon wool columns, which is dominant in the case of B-lymphocytes. This method confirms the fact that T-lymphocytes have a rapid migration and B-lymphocytes a slow migration. We have isolated a third population, having neither the T markers, nor the B markers. It has a very homogeneous migration, centered on the two classes 1,05 and 1,10 mum.sec.-1.V.-1.cm.     

Topology of chromosomes 18 and X in human blastomeres from 3- to 4-day-old embryos.

The positions of chromosomes 18 and X fluorescence in situ hybridization signals were analyzed in blastomeres generated from human in vitro fertilization 3- to 4-day-old embryos after preimplantation screening of aneuploidy of chromosomes 13, 16, 18, 21, 22, X, and Y. Fluorescent signal localization compared with a three-dimensional sphere model of random signal distribution revealed significant differences, providing evidence of peripheral localization of chromosome 18 in aneuploid (p=0.0013) and aneuploid/euploid blastomeres (p=0.0011). No differences were found in localization of chromosome 18 in euploid and in chromosome X in euploid and aneuploid blastomeres.     

Morphologic characteristics of bacteria treated with human blood serum and immunoglobulin

The electron microscopic study of ultrathin sections of bacterial cells treated with blood serum has revealed the presence of the immunoglobulin cover located on the cell wall and consisting of blood serum components. Staphylococci (strain Smith), studied with the use of ruthenium red, have been found to possess the capsule and the immunoglobulin cover located at its base. The immunoglobulin cover has proved to be more pronounced in gram-positive bacterial (actinomycetes, peptococci, staphylococci) and faintly pronounced in gram-negative bacteria (Bacteroides, Proteus, Pseudomonas, Escherichia).     

Microscopic assessment of pronuclear embryos is not definitive

The microscopic classification of embryos, especially unipronuclear embryos, is not very precise. A number of undocumented and unipronuclear embryos were determined to be diploid following karyotyping and fluorescence in situ hybridization (FISH). Accelerated and asynchronous pronuclear dismantling at the time of checking for embryo fertilization accounts for this disparity. Diploid embryos were also observed among tripronuclear embryos. However, not all embryos ascertained as diploid by FISH were karyotypically normal following full karyotype analysis. By taking into account the "background" abnormality rate, the rate of diploid embryo wastage was estimated to be about 40% among undocumented embryos and about 58% in total. A high percentage of misclassification infers an unintended loss of otherwise transferable embryos. Such a discrepancy is particularly important to older women who have fewer embryos. If these are a woman's only embryos, preimplantation genetic diagnosis might be applicable in determining those that are diploid and suitable for transfer. This could potentially reduce the number of wasted embryos and cycles. The present study has also shown that mosaicism is common but it is still unclear whether mosaicism is indicative of embryonic abnormality or is a fairly common phenomenon among healthy embryos. Bipronuclear embryos that present with abnormal or delayed cleavage are often chaotic in their chromosomal constitution. Such embryos should not be transferred.     

The photometric characteristics of the nuclei of normal human and bovine lymphocytes and in pathology]

Chromatin structure has been studied with a method of optico-structural machine analysis for interphase nuclei of peripheral blood lymphocytes in healthy donors, a girl with the Shereshevski?-Turner syndrome (45, XO) and her parents, and for peripheral blood lymphocytes of a cow, Bos taurus L., in the last trimester on the background of preeclamptic syndrome and with normal cow pregnancy of compared terms. A significant change was revealed both in human heteroploidy and in developing preeclamptic syndrome in animals as concerns such indexes as the nucleus area, integral optical density, the amount and area of condensed, decondensed chromatin. The profile of histograms, according to the given parameters, is distinctly changing. The analysis based on constructing a two-measured field in the given coordinate system showed changes in the lymphatic population structure with the increase in the rate of "heavy", enriched DNA cells.     

Tecto-thalamo-telencephalic visual system of the brain in 13-day-old chick embryos

Light and electron microscopic studies have been made of the nervous tissue in three parts of the tecto-thalamo-telencephalic visual system--i.e. tectum opticum, nucleus rotundus of thalamus and ectostriatum of telencephalon--of 13-day chick embryos. Neuroblasts and neurones at various stages of differentiation were described together with various types of synaptic and nonsynaptic intercellular contacts in the neuropil of these brain structures. Heterochronous maturation of these parts of the visual system in embryogenesis was noted which reflects the level of their phylogenetic maturity. Being phylogenetically more ancient structures, tectum opticum and nucleus rotundus reveal differentiation earlier than ectostriatum which is phylogenetically younger.     

Ratio of concentration of ions of uni- and bivalent metals in the tissues of developing chick embryos and 1-day-old chickens]

The ratio of concentrations Na+/K+ decreases in the brain and liver and increases in the cardiac muscle during the chick embryogenesis. The maximum concentration of Ca2+ was noted in the tissues under study on 14th day of chick embryonic development and its content decreased reliably at the subsequent stages. The concentration of Mg2+ in the tissues under study decreased during the embryogenesis as well.     

Estradiol secretion by the ovary of 19-day-old hypophysectomized and sham-operated chick embryos

The amounts of estradiol released into culture media by ovaries from 19-day-old hypophysectomized (decapitated) and sham-operated chick embryos were determined by RIA. Per single ovary, ovaries from decapitated embryos secreted slightly less estradiol than ovaries from sham-operated ones during a 4-h culture period (837 +/- 413 vs 935 +/- 378 pg), but this difference was not significant. On an ovarian weight basis, ovaries from decapitated embryos secreted slightly more estradiol than ovaries from sham-operated ones (1.15 vs 0.91 ng), but this difference was not significant either. It is concluded from these results that the hypophysis does not control estradiol secretion by the ovary in the 19-day-old chick embryo.     

Morphologic and enzymatic characteristics of healthy human kidney epithelial tubular cells in culture

The present study defines morphological and enzymatic characteristics of human healthy renal tubular cells in primary culture, and during three passages. The results confirm the ability to culture human renal tubular cells and to quantitatively evaluate the activity of specific tubular enzymes in the cells and in the culture medium. Such determinations can be compared to similar enzyme activity research in renal biopsies or in urines, to investigate human renal cell functions, in clinical, pharmacological or toxicological approaches.     

Synthetic profiles of polypeptides of human oocytes and normal and abnormal preimplantation embryos

There is considerable variation in the rate of development in vitro of individual preimplantation human embryos. The relationship between the rate of development and patterns of polypeptide synthesis in individual embryos was examined using SDS-PAGE and autoradiography. After incubation in [35S]methionine, 19 polypeptide bands were identified that change between fertilization and the morula stage. Although changes in two of the bands occurred in embryos that were developing normally and in ageing oocytes, and are thus independent of fertilization, the changes identified in the remaining 17 bands occurred only after fertilization. In embryos that were developing abnormally, as assessed by delayed cleavage, cleavage arrest or extensive fragmentation, the alteration in polypeptide synthetic profiles increased with increasing abnormality.     

A cytogenetical study of 7-day-old bovine embryos of poor morphological quality

Seven-day-old embryos were collected from Canadian Holstein and Ayrshire heifers after superovulation with pregnant mare's serum gonadotropin (PMSG) or follicle-stimulating hormone (FSH). A total of 103 morphologically abnormal (type C) and 23 morphologically normal (type A) embryos were cytogenetically analyzed after 4, 20-24, or 44-48 h of culture in enriched phosphate-buffered saline or Eagles minimum essential medium. Twenty-one of 23 (91.3%) type A and 75 of 103 (72.8%) type C embryos had cells in metaphase. Among the 21 type C embryos produced by PMSG stimulation, 17 (80.9%) could be analyzed: 6 were mixoploid (two 2n/3n, three 2n/4n, one 2n/6n), 2 were aneuploid (61 XXY), and 9 were diploid. Among the 82 type C embryos produced by FSH stimulation, 58 (70.7%) could be analyzed: 6 were mixoploid (one n/2n, one 2n/3n, three 2n/4n, one 2n/4n/8n), 2 were polyploid (4n), and 50 were diploid. No abnormalities were observed in the type A embryos.