Sites of serum alpha-fetoprotein synthesis in the human and in the rat.

Selected tissues from human embryos of 6 to 9 weeks' gestation, from rat fetuses of 15 days' gestation, and from rats 2 days of age were incubated with (14)C-labeled amino acids. Immunoelectrophoresis of the culture fluid after incubation, using rabbit antisera against human and rat fetal serum proteins, followed by radioautography revealed that: 1) Radioactive alpha-fetoprotein was present in cultures of human liver, rat liver, and rat yolk sac, but not in cultures of human or rat brain, lung, heart, kidney, intestines, skeletal muscle, skin, or placenta; human yolk sac was not studied. 2) Radioactive transferrin was also present in rat yolk sac cultures, and the same protein was found in rat liver cultures as well. 3) Rat liver and rat placenta cultures both produced radioactive serum Ralpha(2)-globulin. Serum alpha-fetoprotein concentrations in the rat declined abruptly after birth to approximately half of the prenatal level by 2 to 3 days of age, in accord with the loss of the fetal membranes at delivery; the alpha-fetoprotein level then remained relatively constant until the rat was 6 to 8 days of age, after which synthesis of the protein was increasingly suppressed.

Development of gamma G, gamma A, gamma M, beta IC-beta IA, C 1 esterase inhibitor, ceruloplasmin, transferrin, hemopexin, haptoglobin, fibrinogen, plasminogen, alpha 1-antitrypsin, orosomucoid, beta-lipoprotein, alpha 2-macroglobulin, and prealbumin in the human conceptus.

The synthesis of gammaG, gammaA, gammaM, beta(1C)/beta(1A), C'1 esterase inhibitor, ceruloplasmin, transferrin, hemopexin, haptoglobin, fibrinogen, alpha(1)-antitrypsin, orosomucoid, beta-lipoprotein, alpha(2)-macroglobulin, and prealbumin was studied in 15 normal human embryos and fetuses of 29 days to 18 wk gestation and in the yolk sacs of four embryos from 5.5 to 11.5 wk gestation using tissue culture in (14)C-labeled amino acids followed by radioimmunoelectrophoresis. The human embryo as early as 29 day gestation synthesized beta(1C)/beta(1A), C'1 esterase inhibitor, transferrin, hemopexin, alpha(1)-antitrypsin, beta-lipoprotein, alpha(2)-macroglobulin, and prealbumin in culture. At 32 days gestation ceruloplasmin and orosomucoid were also synthesized, but synthesis of fibrinogen was not observed before 5.5 wk. Synthesis of gammaM occurred as early as 10.5 wk gestation, and gammaG synthesis was found in cultures as early as 12 wk gestation; gammaA synthesis was not detected in any of the tissue cultures. With the exception of the gamma-globulins, each of the proteins studied was synthesized by the liver, but additional sites of synthesis for some of these proteins were also found. Synthesis of gammaG and gammaM occurred primarily in the spleen, but other sites of synthesis were noted as well. Changes in the concentrations of most of these proteins and plasminogen in embryonic and fetal serum from 5.5 to 41 wk gestation, in amniotic fluid from 6.5 to 38 wk gestation, and in the sera of neonates during the 1st 3 wk postpartum are described. Although gammaA, gammaM, ceruloplasmin, or haptoglobin were not detectable in some of the embryonic and fetal sera, gammaA and ceruloplasmin were both present as early as 6.5 wk gestation, haptoglobin by 9.5 wk gestation, and gammaM by 17 wk gestation. Each of the other proteins were present in all of the sera examined.

A protein from polymorphonuclear leukocytes (LEM) which affects the rate of hepatic amino acid transport and synthesis of acute-phase globulins.

A proteinaceous secretion from phagocytizing polymorphonuclear leukocytes, termed "leukocytic endogenous mediator" (LEM), has been shown to have marked effects on hepatic amino acid transport and RNA and protein synthesis. A single injection of LEM results in a marked accumulation of labeled nonmetabolizable model amino acids in the liver of normal rats. The LEM-stimulated uptake of amino acids by liver was observed in adrenalectomized, hypophysectomized, thyroidectomized, or diabetic rats and could not be duplicated by pharmacological doses of a large variety of hormones. In addition, LEM stimulated an increased uptake of alpha-aminoisobutyric acid by isolated livers during their perfusion in vitro. LEM also stimulated an increased incorporation of orotic acid into hepatic RNA of intact rats, especially into the bound ribosomal fraction. This increased synthesis of RNA preceded an enhanced hepatic production of a number of the acute-phase plasma globulins. LEM did not stimulate the adenylate cyclase-cAMP system in liver and was not found to utilize this system as a second messenger. Thus, the effects of LEM in stimulating hepatic amino acid transport appear to be direct, without mediation by other hormones, and to be independent of cAMP. On the other hand, the ability of LEM to stimulate RNA and acute phase globulin synthesis in liver may require the presence of physiological quantities of hormones such as adrenal corticoids.

Course of serum-Ig concentrations in B12 chickens of the UM line.

The University of Munich chicken line ("UM") is isogenic in respect to the MHC and divides into normogammaglobulinemic, permanent and transient dysgammaglobulinemic individuals. Hence the immune defect is independent of the MHC. Continual analysis of the immunoglobulins until the 50th week of life revealed: one group of dysgammaglobulinemic individuals showed an initial IgG peak between the third and sixth week of life. Unusually high IgM and IgA levels occur in permanent and transient dysgammaglobulinemic individuals previous to the appearance of the IgG deficit and previous to a possible initial IgG peak. These high levels remain throughout the life of the chicken, possibly due to a missing negative feedback mechanism. Transient dysgammaglobulinemic chickens also exhibited increased IgM and IgA values after IgG normalization. Based upon our results, we postulate that the dysgammaglobulinemia defect is already preprogrammed during late embryonic development. The prevalence of a B or T-cell defect is still under discussion.

[Biochemical study on the 1st case of analbuminemia in France]. / Etude biochimique du premier cas d'analbuminémie en France

A new case of analbuminemia was described as follows for a six month's old child of Algerian origin. The discovery of the disease was made by chance, the clinical signs were limited to small oedema. The serum albumin concentration was 64 mg/1 and its immunochemical action was identical to that of normal albumin. The system reacted by an increase of the synthesis of globulins. For the subject, the alpha1-antitrypsin, ceruleoplasmin, hatoglobin, alpha2-macroglobulin, transferrin, immunoglobulins M contents were three times higher than the standard figures. The analysis of the distribution of non esterified fatty acids ususally carried by albumin was normal. On the other hand, it was possible to show that the presence of free bilirubin independant from proteins could be detected for a concentration of 17 micronmol/l. A study of the family showed a standard repartition of albumin and globulins. The genetic origin observed in the symptoms was confirmed by the consanguinity of the parents.

[Use of molecular biology approaches in studying hereditary diseases]. / Primenenie molekuliarno-biologicheskikh podkhodov pri izuchenii nekotorykh nasledstvennykh boleznei.

The application of some molecular biological methods for studying the causes of hereditary diseases induced by quantitative changes of the normal protein synthesis is discussed. The group of hereditary anemiae in humans (alpha- and beta-thalassaemia) taken as an example, possible defects at various stages of the protein synthesis control and modern methods of the analysis of these defects are considered and promises offered by such approaches are shown.

[Biosynthesis and cellular localization of betal-g-globulin in the human placenta]. / Izuchenie biosinteza i kletochnoi lokalizatsii betal-g-globulina v platsente cheloveka

Pregnancy-specific beta1-globulin (beta1-GP) described by Tatarinov and Masyukevich, 1970) was shown by means of immunodiffusion in agar gel to be similar to specific pregnancy beta1-glycoprotein of Bohn (1971) as well as to pregnancy associated plasma protein-C of Lin et al (1974). It was found that in immature placenta (6-12 weeks of pregnancy) incorporation of 14S-amino acids into beta1-GP took place. Studies on immunofluorescence showed that beta1-GP was localized in cytoplasma of trophoblastic cells of placenta chorion. Beta1-GP production apparently begins in Langhans cells and proceeds in syncytiotrophoblastic cells, but at lower activity and secretory scale.

Vascular permeability factor of Pseudomonas aeruginosa.

The production of vascular permeability factor (PF) by certain strains of Pseudomonas aeruginosa has been demonstrated in rabbits injected intradermally with culture filtrates followed by intravenous injection with Pontamine Sky Blue 6BX. The dose-response curve was found to be rectilinear when lesion diameters, within the range of 10 to 20 mm, were plotted against log dose. Thus, PF in test filtrates can be measured with reasonable accuracy by the concomitant testing of a reference PF. In contrast to the titers of PF obtained with Vibrio cholerae cultures, those with strains of P. aeruginosa were rather low. Thus far, PF has been demonstrated only in shallow still cultures of P. aeruginosa and not in shake cultures. A variety of commercial media were tested for the production of PF, but none was satisfactory. A synthetic medium that gave more reproducible and higher yields of PF was developed. Cultivation at 30 C generally gave higher yields of PF than at 37 C. PF was destroyed by heating at 60 C for 30 min or by digesting with trypsin or Pronase. Strains producing larger amounts of PF appeared to have greater virulence when inoculated onto the surface of burns in mice than those yielding little or no PF.

Electrophoretic pattern of proteins in lepromatous leprosy.

Serum total proteins and the various protein fractions were studied in fifty cases of lepromatous leprosy and in eleven cases of lepromatous leprosy with lepra reaction. The study revealed a significant increase in serum total proteins in both lepromatous leprosy and lepra reaction groups, when compared with normal healthy subjects. The percentage rise was found to be 14.5% and 22.95% for lepromatous leprosy and lepra reaction respectively. The globulin fraction showed a significant elevation, while albumin showed a decrease. Thus a reversal of A/G ratio was observed in both the disease groups. Alpha-1 and Alpha-2 globulins were found to be significantly increased in both the disease groups. Beta globulins did not reveal any significant alteration. It was interesting to note the presence of an additional globulin fraction in seventeen patients of lepromatous leprosy and two cases of lepra reaction. Gammaglobulin showed a significant rise in lepromatous leprosy (56.16%) and in lepra reaction (60.72%). The significance of the above findings are discussed in the light of available literature.