Sulphonated phthalocyanine induced caudal malformative syndrome in the chick embryo.

Sulphonated phthalocyanine (Pht.) has been tested for its possible noxious effect on the developing chick embryo. When injected into the subembryonic cavity of 40-45 hours incubated chick embryos (mainly 10-20 somite pairs), Pht. induces a highly reproducible caudal malformative syndrome (trunk and taillessness, various anomalies of the limbs). The main effect is--in about 15% of the malformed specimens--associated with unilateral microphthalmy and, less frequently, with coelosomy. Microscopically developmental disturbances of the caudal axial organs, of the mesonephros and of the limbs are observed. The initial pathological changes, at microscopic level, are necrosis and hemorrhages in the caudal axial and paraxial area. The allantois is poorly developed or even absent. Skeletal changes involve anomalies of the ribs and of the vertebral column and total or partial absence of the pelvic girdle bones. The high mortality, mainly during the first week, is due--first of all--to the developmental disturbances including the poor development or absence of the allantois. Control experiments with CuCl2 suggest the ethiological role of Cu. Pathogenetic aspects are discussed.

The role of the cerebrospinal fluid during embryonic development. A biochemical study.

The total protein content and the protein fractions were determined in the blood serum and the cerebrospinal fluid (CSF) of chick embryos and foetuses between the 4th and 20th day of incubation and of 2-day-old chickens (by Lowry's method and by polyacrylamide gel electrophoresis respectively). The main results were the following: The total protein content of the CSF increased from 1.8 mg/ml on day 4 to 3.1 mg/ml on day 10. After a slight decrease to 2.6 mg/ml on day 12 no further changes were detected. In the blood serum the total protein content was 4.3 mg/ml on day 4 and 4.7 mg/ml on day 9. Beginning with day 10 it increased constantly to 23.2 mg/ml on day 2 after hatching (daily rate 1-1.2 mg/ml). It results that the "protein barrier" between blood and CSF begins to function after 10-12 days of incubation. Of the protein fractions determined, the light fraction I in the CSF represents 50 per cent of the similar blood serum fraction (on day 10). As to the heavy fractions XI-XIV, their concentration in the CSF represents, beginning with day 10, 51 per cent of the similar blood serum fraction (until the end of the period investigated). It results that the "protein barrier" has an active, selective character. Some comparative aspects with literature data are discussed.

[Dynamics of the action of cyclophosphamide. Studies in the chick embryo]. / Uber die Wirkungsdynamik von Cyclophosphamid. Untersuchungen am Hühnerembryo.

The dynamics of the Cyclophosphamide (CPA) effect was investigated in the chick embryo by a vital fluorochroming method worked out previously. In the 4-day-old embryo the morphological changes (at the level of light microscopy) in the limbs and in the mesencephalon induced by 100 microgram CPA appeared after a period of latency of about 10 hours. Successive blood control in the same embryo, treated with 50 microgram CPA, revealed three consecutive modifications of the blood cells: a great number of abnormal mitoses, the appearance of a great number of multinucleated, nonviable cells and the recovery of the lost cells. No difference could be detected between the effect of freshly prepared and stored CPA solutions. The fate of CPA (a bifunctionally alkylating agent) in contact with living tissues is discussed.

[Vital fluorochroming showing the effect of cyclophosphamide on the embryo]. / Vitalfluorochromierung als Mittel zur Auswertung der Cyclophosphamidwirkung beim Embryo.

Besides the usual methods, vital fluorochroming may globally show the extent of the cytotoxic effect of Cyclophosphamide (CP). By intravital treatment with highly diluted fluorochromes (acridine-orange, etc.) necrobiotic and dead cells, cell debris and phagosomes appear electively fluorescent. Glycosaminoglycans and various mucopolysaccharides show a somewhat weaker fluorescence. Autopods and the mesencephalon were selected for comparative assessment of the effect of other cytotoxic agents. Important results were also obtained by the repeated examination of blood (erythrocytes) and of hemopoetic foci. Cytotoxic effect within the embryo, teratogenesis and chemotherapeutic action are strongly correlated.

[Perichordal necroses. Contribution to the understanding of the spinal column morphogenesis]. / Perichordale nekrosen. Ein Beitrag zum Verständnis der Morphogenese der Wirbelsäule.

In 12--15 days old rat embryos and 12 days old mouse embryos vitally stained by acridin orange, a perichordal necrosis appears, showing metameric intensifications at the level of the dense sclerotomites. Similar aspects were detected in serially sectioned rabbit and human embryos (6--7 mm c.r. length). The authors correlate the necrotic zones mentioned with the inductive functions of the notochord.

[Fluorescence microscopy on embryonic and fetal cell degeneration]. / Fluoreszenzmikroskopische Untersuchung embryo-fötaler Zelldegenereszenzen.

Fluorochromes (acridin orange, quinakrin, koriphosin, auramin) introduced into the embryonic or foetal blood stream induce in the normal areas of cell death an elective fluorescence, as attested by experiments carried out on chick and rat embryos and foetuses. This vital method may serve as an experimental model for investigations concerning the transfer from the mother to the embryo or foetus of teratogenic substances as well as of their persistance and distribution--under certain experimental conditions--within the organism of the embryo or foetus.

Researches on the macrophage reaction in embryonic necrotic foci by means of cell cultures.

Macrophage transformation of the fibroblasts and the attraction of macrophages in cell cultures of chick embryo have been investigated. Cultures were treated with choline hydrochloride, DNA, lactalbumine hydrolysate, lysine, peptone and ATP. An intense agglomeration of macrophages in the contact area with the agar gel containing choline could be observed. Cultures treated with DNA have a great number of macrophages together with a numerous fibroblast population. In cultures treated with the other substances, macrophages also appear, yet their number is not so great. The results obtained plead for a triggering of the macrophage reaction and of the necrosis in embryo tissues and organs by substances identical or similar to those used in our cultures.