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Fetal proteins in various tumors.

Radial immunodiffusion assay was used to measure fetal hemoglobin (HbF) concentrations in 312 patients with various malignancies. In 305 of these, alpha-fetoprotein (AFP) was measured by radioimmunoassay. The concentration of HbF exceeded 3 SDs above the normal mean in 68 of 312 patients, most notably in patients with leukemia, multiple myeloma, lymphoma, bladder carcinoma and testicular tumors. HbF was correlated with total hemoglobin concentration and with serum AFP concentration in hepatoma and bladder carcinoma.

Carcinoma

A fetal protein in chromatin of Novikoff hepatoma and Walker 256 carcinosarcoma tumors that is absent from normal and regenerating rat liver.

Antibodies to chromatin proteins of Novikoff hepatoma cells formed precipitin bands in the double-diffusion immunoprecipitation assay with chromatin proteins of Novikoff hepatoma, Walker 256 carcinosarcoma, and 18-day fetal rat liver. The antigen used for preparation of antiserum was the chromatin proteins initially extracted with 3 M NaCl-7 M urea and soluble after dialysis to 0.14 M NaCl-0.35 M urea. The chromatin proteins used for analytical studies were extracted with 0.6 M NaCl containing 0.01 M Tris-HCl (pH 8) and 100 muM phenylmethylsulfonyl fluoride. Corresponding chromatin proteins of normal and 18-hr regenerating rat liver, heart, and kidney did not form precipitin bands. The antigen was purified from the chrmatin of Novikoff hepatoma cells by exclusion chromatography on Sephadex G-150 and preparative nondenaturing polyacrylamide gel electrophoresis. Its migration on denaturing sodium dodecyl sulfate-polyacrylamide gels corresponded to a molecular weight of 26,000. Amino acid analysis showed that the ratio of acidic to basic amino acids was 1.4 to 1.0. Evidence for its homogeneity included its migration as a single protein spot on two-dimensional polyacrylamide gel electrophoresis and its single lysine amino-terminal amino acid. This protein is a glycoprotein, as shown by the presence of 15 moles of galactosamine per mole of antigen. These studies demonstrate the presence of a fetal glycoprotein in the chromatin of two tumors that may have an important role in determining their gene products.

Animals

Serum alpha fetal protein in a three year old child with hepatoma.

A 3 year old child with primary hepatocellular carcinoma and high AFP concentrations is described. Following hemihepatectomy, a sharp decrease and return to normal of serum AFP concentrations indicated the completeness of the surgical procedure. Repeat-normal serum AFP concentrations (less than 19 ng/ml), found during a three year follow-up, correlated well with the absence of clinical, laboratory and x-ray evidence of tumor recurrence. The differential diagnosis of abnormal AFP concentrations in childhood is discussed, and the importance of the AFP assay in the follow-up of post-hemihepatectomy patients for the assessment of the completeness of the surgical procedure, the prognosis, and the early detection of tumor recurrence is stressed.

Carcinoma, Hepatocellular

Carcino-fetal proteins and gastric cancer: the site of alpha-fetoprotein synthesis in gastric cancer.

Although several investigators have reported serum alpha-fetoprotein positive gastric cancer with or without metastasis to the liver, its site of production is still unclear. We studied on the site of alpha-fetoprotein synthesis in our cases which showed remarkably high level of serum alpha-fetoprotein, and clarified the presence of alpha-fetoprotein producing gastric cancer by means of direct immunofluorescent technique. However, we also knew the hepatocytes adjacent ot the metastatic lesions could also synthesize alpha-fetoprotein, although these hepatocytes were not known whether under regeneration or degeneration. Through this study, the other carcinofetal proteins such as carcino-placental alkaline phosphatase and CEA were examined using the sera or tumor tissues. Our results will support the idea that cancer is the disease of cell differentiation.

Adenocarcinoma

[Study of soluble proteins from human amniotic fluid by fraction chromatography (author's transl].

The nature and origin of soluble proteins from human amniotic fluid have been investigated by gel filtration chromatography on Sepharose 6B and ion-exchange chromatography on Ecteola-cellulose. Each fraction has been studied by immunoelectrophoresis. Specific antisera against serum proteins, amniotic fluid proteins, fetal proteins and salivary proteins have been used. These various antisera showed that the amniotic fluid contains maternal proteins, but also more specific proteins from fetal origin such as beta2-microglobulin, urinary mucopolysaccharides, salivary proteins and carcinoembryonic antigen. These results not only confirm that amniotic proteins are essentially from maternal origin, but prove the important fetal contribution and emphasize the importance of the fetoplacental unit in the monitoring of high risk pregnancies.

Amniotic Fluid

Relationship of maternal amino acids and proteins to fetal growth and mental development.

The relationship of maternal amino acids, proteins, and certain other factors to fetal development (weight and cranial volume at birth and mental and motor scores at 8 months of age) was investigated in 129 gravidas. Maternal prepregnancy weight was significantly related to birth weight and cranial volume in the newborn. Among amino acids, glycine, lysine, and total amino acids were positively correlated with birth weight, whereas valine and threonine showed a negative correlation. Cranial volume at birth was significantly related to the level of glycine and alpha1 globulin. Other protein and amino acids which contributed to cranial volume included threonine, histidine, glutamic acid, and beta globulin, negatively correlated, whereas glutamine was positively correlated. Alpha1 globulin (negatively) and glycine (positively) made significant contributions to motor development. For mental development, alpha 1 globulin and histidine showed a negative correlation, whereas isoleucine was positively correlated. Findings indicated that the concentrations of certain maternal amino acids and proteins in the third trimester of pregnancy correlated significantly with fetal growth and development. This study suggests: (1) specific proteins or amino acids may be responsible for different developmental measures; (2) injudicious dietary restrictions in pregnancy should be avoided; (3) the determination of alpha1 globulin and a few amino acids such as glycine, lysine, and histidine in late pregancy may be used as predictors of fetal growth and development.

Alpha-Globulins

The ontogenesis of calcium-binding protein in fetal rat kidney.

The presence of calcium-binding protein (CaBP) in the nucleus and cytoplasm of epithelial cells of renal tubules varies with the physiologic state. As part of a study to determine whether or not intranuclear CaBP precedes intracytoplasmic CaBP in the same cell, we used peroxidase-labeled antibody against human renal CaBP to localize CaBP in fetal rat kidney tubules. This paper reports examination of kidneys from rats on each day of gestation from the 10th to term (21 days) and on each of the first seven postnatal days. CaBP was first detected in fetal rat kidneys on the 19th gestational day. The histochemical staining reaction that revealed the CaBP was less intense than that produced in kidneys from adult animals, but its distribution was like that in adults, with some cells having no CaBP, others having it in the cytoplasm only, in the nucleus only, or in both. By the seventh postnatal day the staining reaction was similar to the adult patterns in all respects.

Animals

Fetal specific serum proteins of the Indian buffalo.

Fetal specific serum proteins of the Indian buffalo were investigated using rabbit antibuffalo fetal serum, absorbed with lyophilised adult buffalo serum, and subjected to gel diffusion and immunoelectrophoretic analysis. AFP was found to be the major fetal specific serum protein of the Indian buffalo. A close antigenic relationship between the fetal specific serum proteins of the buffalo, sheep and goat and a distant relationship between these proteins of ruminants and man was found by immunological cross reaction studies.

Animals

The use of alpha fetoprotein in prenatal diagnosis.

Maternal or blood levels of alpha fetal protein are elevated in the presence of certain fetal anomalies, particularly those of the neural tube in which protein from the cerebrospinal fluid may leak into the amniotic fluid. Since this is a nonspecific elevation, other diseases are potentially screened for by this determination. Alpha fetal protein levels should be measured on any amniotic fluid samples obtained during the middle trimester of pregnancy.

Amniotic Fluid

Glucocorticoid receptors in lung. Comparison between nonactivated and activated forms of the cytoplasmic glucocorticoid binding protein and their relationship to the nuclear binding protein of fetal rabbit lung.

In the absence of salt the cytoplasmic glucocorticoid receptor of fetal rabbit lung sediments at 7 S while the nuclear receptor sediments at 4 S. However, if nuclear extracts are mixed with receptor-depleted cytosol preparations in dilute buffer solutions without added salt, the nuclear 4 S receptor sediments as a 7 S species similar to that observed for the cytoplasmic form under the same conditions suggesting an interaction of the nuclear receptor with other cytosol proteins rather than with itself. In addition, both cytoplasmic and nuclear receptors sediment at 4 S in 0.4 M KCl and a major fraction of the nuclear receptor has an agarose elution profile identical to that of the cytoplasmic receptor. Thus a major fraction of the nuclear receptors is indistinguishable from the cytoplasmic receptors by the methods used. Since the cytoplasmic receptor sediments at 4 S in 0.15 M KCl, it is suggested that in vivo the glucocorticoid receptor may exist as a 4 S species and that the 7 S form described previously may result from an interaction of the 4 S component with other cytosol proteins in hypotonic media. About 25% of the receptor present in nuclear extracts has an agarose elution profile different from that of the cytoplasmic receptor in 0.4 M KCl. This suggests that either the nuclear receptor associates with itself or other nuclear proteins or that more than one form of nuclear receptor exists. Earlier observations suggested that in the absence of hormone the glucocorticoid receptor is localized exclusively in the cytoplasm of lung cells and that the nuclear receptor is formed by a transfer of the cytoplasmic steroid-receptor complex into the nucleus. A prerequisite for this transfer seems to be a modification of the receptor to an active form which can bind to nuclei. This receptor transfomration, referred to in this paper as activation of the receptor, can occur in the absence of nuclei and is highly dependent on temperature and ionic strength. Cytoplasmic receptors activated either by heating or by exposure to high ionic strength are indistinguishable from nonactivated receptors by sucrose density gradient analysis or by agarose gel filtration in solutions containing 0.4 M KCl. Simiarly, no significant difference in the absence of salt is observed after activation by heating. These results suggest that activation of the cytoplasmic glucocorticoid receptor involves conformational changes which favor its transfer and/or binding to nuclear sites rather than conversion of a 4 S species to a faster-sedimenting form by dimerization or by addition of another protein unit as has been proposed for the activation of the estrogen receptor of the rat uterus.

Animals

Light microscopic and ultrastructural observations in advanced stages of induced exencephaly and spinal bifida.

This investigation was performed to demonstrate the morphologic basis of the elevation of fetal proteins in the amniotic fluid of fetuses with neural tube defects. Pregnant rats were treated with hypervitaminosis. A to induce exencephaly or with trypan blue to produce spina bifida aperta. The malformations were studied on days 15-20. On day 15 of gestation, edema developed in the primitive nervous tissue. This was followed by the appearance of quickly expanding hemorrhages throughout the ventricular and intermediate zones. Some capillaries did not rupture but collapsed and showed degenerative changes of the endothelium, probably due to lack of blood perfusion. The ventricular layer in exencephaly and spina bifida aperta was exposed to the amniotic cavity due to non closure of the neural tube. On day 17, this superficial lining of the primitive nervous tissue was disrupted by the expanding hemorrhages and subsequent necrosis. As a result vast amounts of fetal blood and cell debris were extruded into the amniotic fluid. During days 18 to 20, the degeneration of the nervous tissue proceeded rapidly. This process showed the same features in the ventricular cells, the primitive neurons and the neurons. Initially it was characterized by condensation of the nuclear chromatin and the cytoplasm, irregular outlines and breakdown of the plasma membrane. Only part of the cell debris was phagocytozed by macrophages. It is concluded that the leakage of fetal serum and cell debris causes the elevation of fetal protein levels in the amniotic fluid of fetuses with open neural tube defects.

Abnormalities, Drug-Induced