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Saline extracts from human lenses were studied during prenatal eye development. Using immunodiffusion and immunoelectrophoresis with a specific antiserum against total extract of human adult lenses and monospecific antisera against the individual crystallin classes, it was established that alpha and beta crystallins were present in extracts from eye lenses from 5-week-old embryos while gamma crystallins could be detected in extracts from embryonic lenses at the beginning of the 6th gestational week. When analyzed by isoelectric focussing all alpha and beta crystallin components were found in extracts from 8-week-old embryos, while those of gamma crystallins were shown to appear successively within the period between the 8th and 14th gestational week. Some changes in the relative proportion of the different crystallin classes and subclasses were observed in the course of prenatal eye lens development between the 8th and 26th gestational week, as the percentage of alpha, high molecular weight and gamma crystallins was increased while the proportion of beta crystallins was gradually lowered.

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Serum samples from healthy subjects of different ages (within the age range 1-75 years) were tested for the presence of antielastin IgG, IgM, IgA and IgD by an enzyme-linked immunosorbent assay, utilizing insoluble human aortic elastin. All the tested sera showed detectable levels of antielastin antibodies of the four classes with the following regularities in changing their level with age. Antielastin IgG and IgM showed relatively high levels in the serum of children, growing even higher in the serum of subjects 18-20 years old. Then their levels were stabilized in the serum of 30-60 year olds for IgG and of subjects 30-50 years old for IgM and gradually decreased thereafter. The antielastin IgA showed non-significant changes with age up to the age of 40 and then its level gradually increased. The antielastin IgD showed statistically non-significant changes with age and a tendency to decrease after the age of 60.

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The acquired immunodefficiency syndrome (AIDS) is characterized by a gross defect in the cell-mediated immune response. However, infection with human immunodeficiency virus (HIV), which is the generally accepted etiological factor of AIDS, cannot explain by itself the following problems: why do not some of the seropositive subjects develop AIDS or AIDS-related complex; why are some of the patients with AIDS seronegative for HIV and its corresponding antibodies; what is the reason why some of the healthy seronegative subjects from groups at a high risk for AIDS (homosexuals, hemophiliacs and drug abusers) have low T-helper to T-suppressor ratios. We suggest that some additional factor is necessary for the development of AIDS. We propose that the factor needed is a 'partial functional thymectomy'. We suspect that slow fungal infections, producing thymotoxic metabolites, may be a major cause for the latter.

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The conjugates beta-amanitin-concanavalin A and phallacidin concanavalin A were tested for direct cytotoxicity on L1210 lymphocytic leukemia cells by a combined in vitro-in vivo bioassay. Both conjugates exerted strong direct cytotoxicity on the tumour cells.

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A cytogenetic investigation was carried out of 55 workers from the petroleum-processing industry and of 30 control subjects. The frequencies of both structural and numerical chromosome aberrations and of sister chromatid exchanges were determined simultaneously and the relationship among these cytogenic indices were analyzed. The incidence of chromosome aberrations and sister chromatid exchanges among the petroleum workers was higher than that among the controls. The cytogenic indices demonstrated a clear dependence on the working environment of the subjects and a correlation with the existence of some reproductive failures in the families of the exposed workers. Our results indicate the presence of some mutagenic risk in the working environment of some of the divisions in the petroleum-processing industry due to the genotoxic influence on the somatic cells of the exposed workers at a cytogenetic level. This potential risk is mainly associated with the presence of some heavy oil-fractions and polycyclic aromatic hydrocarbons in the working environment.

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Alpha-elastin of human aorta was investigated by an enzyme-linked immunosorbent assay (ELISA) on the material isolated from aborted human fetuses and healthy subjects killed by accident, assigned to 7 age groups. Samples up to the age of 55 were taken only from regions without detectable changes in the arterial wall, in the 60-75-year age group--both from normal areas and atherosclerotic plaques of the same aortas. An immune serum against alpha-elastin isolated from the normal aortic areas of a 61-year-old subject was produced in sheep. Testing with this serum showed the existence of some antigenic changes in the elastins from different age groups. A prevalence of the species-specific antigenic determinants was observed in alpha-elastin from fetal aorta while in alpha-elastin from the atherosclerotically altered human aorta the cross-reacting antigenic determinants prevailed in its antigenic structure.

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Upon simultaneous administration lysine-orotate increases (about 40-fold) the toxicity in mice of a crude Amanita phalloides extract. This effect, though less prominent, is also observed if these two compounds are injected 1 hr apart. The potentiating effect of lysine-orotate is dose dependent and neither L-lysine nor orotic acid exert any effect on the toxicity of the crude A. phalloides extract. Lysine-orotate increases the toxicity of amatoxins (alpha-amanitin) only, not affecting the toxicity of phalloidins. Thin layer chromatography on silica gel plates and column chromatography on Sephadex LH-20 has proven the formation of a relatively stable complex of amanitin and lysine-orotate. The results demonstrate that lysine-orotate should not be used as a hepatoprotective agent in cases of Amanita intoxication.

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Healthy subjects aged between 25 and 60 (20 cases) and between 61 and 65 (5 cases), and diabetic patients with vascular damage, aged between 24 and 62 (6 cases), were tested by a new method for the detection and identification of elastin-antielastin circulating immune complexes (CIC) in human sera. Such immune complexes were found in all patients' sera and only in one of the controls (at the age of 65). Among different patients, the elastin-antielastin CIC varied in size and elastin content, showing some correlation between these two characteristics and the existence of microvascular complications, as proved by the clinical and paraclinical investigation of the patients.

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The level of the circulating elastin-derived peptides (CEDP) in the serum is believed to reflect the activity of the degradation of the elastic structures. This paper reports a new method, based on the 'sandwich' version of the enzyme-linked immunosorbent assay (ELISA), for the detection and quantification of CEDP in human serum. By this method we investigated the age-related changes in their level among healthy subjects within the age range of 1 and 75 years and among atherosclerotic subjects aged 50 to 75 years. The highest level of CEDP was found in the serum of the atherosclerotic patients, and the lowest, among the healthy subjects between 18 and 50 years of age.

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A modified version of the enzyme-linked immunosorbent assay (ELISA), utilising human insoluble aortic elastin, was used for determination of anti-elastin antibodies in serum from normal and atherosclerotic subjects. The age-related changes in their level among healthy persons were investigated. Anti-elastin antibodies were found in all the tested human sera, showing the highest level at the age of 18-20 and the lowest at the age over 60 and especially among atherosclerotic patients. The possible role of the immune system in the turnover of elastin is discussed.

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