Compound heterozygosity for an apolipoprotein A1 gene promoter mutation and a structural nonsense mutation with apolipoprotein A1 deficiency.

Apolipoprotein (apo) A1 plays a central role in the metabolism of HDL. We describe a novel genetic variant of the apoA1 gene identified in a patient with low concentrations of plasma HDL cholesterol. The proband, a 12-year-old Japanese boy, exhibited markedly low levels of both plasma apoA1 and HDL cholesterol. Genomic DNA sequencing of apoA1 genes of the patient showed a compound heterozygosity for an A to C substitution at 27 bp upstream of the transcription start site of 1 apoA1 allele, and a C to T substitution in another allele at residue 84 resulting in aberrant termination. The point mutation at nucleotide position -27 changed ATAAATA of the putative TATA box signal sequence to ATACATA. In addition to this mutation, the patient was heterozygous for a G to A substitution at position -75. Immunoblotting of an isoelectric focusing electrophoresis gel of the proband's plasma showed a trace amount of normal apoA1. No measurable plasma apoA1 and HDL cholesterol in a patient with homozygosity for nonsense mutation at residue 84 has been reported previously. To determine the effects of substitution either at position -27 or -75, plasmids containing the 5'-flanking region of the human apoA1 promoter fused to the CAT reporter gene were constructed and transfected in HepG2 cells. A construct with the A to C substitution at position -27 showed 41. 8+/-4.2%, and G to A substitution at position -75 showed 72.8+/-15. 2% (means+/-SD, n=3) of CAT activities, compared with the wild-type promoter sequence. A construct with the double substitutions at positions -27 and -75 showed only 22.8+/-1.3% (mean+/-SD, n=3) activity relative to the wild type. Our patient is the first case with a TATA box mutation etiologically related to lipoprotein disorders.

Observation of composition in surface monolayers by X-ray scattering spectra caused by crystal truncation and interferences.

X-ray crystal-truncation-rod (CTR) scattering measurements using synchrotron radiation and an imaging plate can reveal the composition of a surface monolayer. Even when the composition is changed in only one atomic layer on the top surface, the X-ray CTR spectrum can change due to the differences in composition. X-ray CTR spectra are greatly enhanced by X-ray interference when a sample is designed properly. In this paper, it is shown by theoretical calculations and experiments for AlAs/GaAs samples grown by MBE that a 1 ML (monolayer)-thick AlAs layer embedded under 10 ML below the surface can enhance the modulation of an X-ray CTR spectrum.

Prenatal diagnosis of congenital lipoid adrenal hyperplasia.

BACKGROUND: There are no published reports of prenatal diagnosis of congenital lipoid adrenal hyperplasia, which is the rarest form of congenital adrenal hyperplasia. CASE: Congenital lipoid adrenal hyperplasia was diagnosed prenatally based on the existence of one affected sibling in the family, the presence of an amniotic fluid cell karyotype of 46,XY, the appearance of normal female genitalia on ultrasonography, relatively low amniotic fluid concentration of 17 alpha-hydroxyprogesterone, low maternal plasma and urinary concentrations of estriol, and a positive response to the dehydroepiandrosterone sulfate loading test. CONCLUSION: Congenital lipoid adrenal hyperplasia can be diagnosed prenatally. Treatment in early infancy can lead to normal mental and physical development.

A case of myasthenia gravis complicated with hyperthyroidism and thymic hyperplasia in childhood.

We report here a case of myasthenia gravis complicated with hyperthyroidism and thymic hyperplasia. The patient was a 13-year-old girl with struma and hyperthyroidism which began at age 12. Two weeks following the initiation of treatment against hyperthyroidism she developed left blepharoptosis, diplopia, and dysphagia, which responded promptly to edrophonium administration. An increase of the anti-acetylcholine receptor antibody was found in the serum. A chest CT showed a large soft tissue mass in front of the ascending aorta, which was proven histopathologically as thymic hyperplasia. The patient underwent an extensive thymectomy and was placed on combination therapy with an anti-thyroid drug, glucocorticosteroid, and an anti-cholinesterase drug. Her symptoms and signs have been well controlled by this treatment. Coexistence of myasthenia gravis, hyperthyroidism, and thymic hyperplasia in childhood have never been documented in literature.

[A case of subacute sclerosing panencephalitis developing 8 years after immunosuppressive treatment for acute lymphocytic leukemia].

A 13-year-old girl developed subacute sclerosing panencephalitis (SSPE) with atypical absence attacks as an initial symptom. Eight years earlier she had been treated for acute lymphocytic leukemia with cytotoxic treatment and radiotherapy, which had resulted in complete remission. She was first treated with an anticonvulsant because the atypical absence attacks and the presence of epileptic discharges on an EEG suggested epilepsy. However, with this mode of treatment the epileptic discharges did not disappear, but periodic high-voltage slow-wave complex discharges were revealed on subsequent EEGs. The antibody titer for measles virus in the cerebrospinal fluid and serum was elevated, confirming the diagnosis of SSPE. SSPE may arise, though rarely, in an individual in an immunosuppressive state due to congenital immunodeficiency or various kinds of malignancies, and also may arise several years after the contraction of measles infection. Our patient, however, lacked a past history of measles infection or immunization, suggesting the possibility that she had contracted measles during or shortly after the course of treatment for ALL.

[A case of systemic lupus erythematosus associated with meningitis, myelitis, and bilateral optic neuritis].

A 16-year-old girl was admitted to our hospital in August 23, 1986, for headache, nausea and low grade fever. Marked increases in immunoglobulin indices were found in the cerebrospinal fluid. When she was 13, she was diagnosed as having SLE and lupus nephritis. On September 9, 1986, she complained of urinary retention, and pathological reflexes were elicited bilaterally. On September 13, she complained of a sudden loss of vision (count fingers) in the right eye which worsened to a visual acuity of light perception over the next 48 hours. A visual evoked response potentials (VEP) to flash stimulation gave a loss of amplitude and an increase in latency. On September 16, she complained of a similar loss of vision in the left eye. Leakage of dye around the left optic disc was found by a fluorescein angiogram on September 26. These results indicated a diagnosis of bilateral optic neuritis. Both visual acuity returned rapidly over the following month following oral prednisolone treatment. Optic neuritis is an exceedingly rare complication in SLE. Although the visual prognosis have been fairly good in the reported cases, some have resulted in various states of blindness. As for etiology of optic neuritis in our patient, ischemic change of optic nerves due to microvasculopathy as well as slight demyelinating process were speculated by the VEP pattern.

The molecular heterogeneity of nonspecific cross-reacting antigen synthesized by tumor cells and granulocytes.

The molecular heterogeneity of nonspecific cross-reacting antigen (NCA) was examined. Metabolically-labeled glycoproteins were precipitated from cell lysates of human tumor cell lines and of normal peripheral granulocytes with antibodies specific for NCA, and analyzed by SDS-PAGE. NCA components synthesized by three tumor cell lines, QGP-1 (pancreas), HLC-1 (lung) and CAOV-2 (ovary) showed slightly different migration patterns on SDS-PAGE, but the molecular weights of their unglycosylated peptides synthesized in the presence of tunicamycin were all found to be 35K. On the other hand, two molecular species of NCA were identified in normal granulocytes: an 80K mature form derived from a 69K precursor peptide and a 58K mature form from a 41K precursor peptide. Upon SDS-PAGE, the migration pattern of the unglycosylated NCA peptides from tumor cells was affected by the presence of 2-mercaptoethanol, while that of the peptides of granulocytes was not. All the NCAs identified in this study possessed antigenic determinants common to carcinoembryonic antigen as well as those unique to NCA. These results suggest that the molecular heterogeneity of NCA observed thus far resulted from diverse glycosylation of the three fundamental molecular forms of unglycosylated peptides: one with a molecular weight of 35K produced by tumor cells and two with molecular weights of 69K and 41K produced by granulocytes.

Chilaiditi's syndrome in schizophrenic patients.

The condition that a part of the digestive tract interposes between the liver and diaphragm has been called Chilaiditi's syndrome. Although it has been reported that this condition had a higher rate of incidence in patients of mental hospitals than in other individuals, there are hardly any enlightening papers of this condition in the domain of psychiatry. Three schizophrenic patients who had this condition are described in this paper. Many factors such as meteorism, the medication of antipsychotic drugs and negative symptoms of schizophrenic patients are considered as risk factors that bring about this condition in them. Psychiatrists should pay more attention to this condition in the chest roentgenograms of patients because this state can possibly develop into more serious conditions such as ileus.

Antigenic alterations of carcinoembryonic antigen involved in the secretion from various human tumor cell lines.

Antigenic variations of carcinoembryonic antigens (CEAs) produced by 8 human tumor cell lines of various organ origins--4 derived from colonic, one from gastric, one from pancreatic, and 2 from lung cancers--were investigated on the basis of reactivity with several monoclonal antibodies recognizing either the peptide or the carbohydrate moiety of the CEA molecule by using a sandwich-type solid-phase radioimmunoassay (RIA). CEAs from all the cell lines revealed nearly uniform reactivities with the monoclonal antibodies recognizing the peptide epitopes, and no difference was observed between the reactivities of CEAs released into culture medium (m-CEAs) and of those extracted from cells (c-CEAs). On the other hand, with each of 4 monoclonal antibodies recognizing the carbohydrate epitopes, the reactivities of CEAs were found to be highly variable from one cell line to another, and a marked difference was detected between the reactivities of m-CEA and of c-CEA in every cell line. In general, c-CEA reacted much more strongly than m-CEA with these antibodies. The reactivities of some of the CEAs with 2 out of 4 antibodies to sugar epitopes increased significantly after treatment of CEAs with neuraminidase. These results suggest that distinct structural changes in the carbohydrate moiety may occur during the releasing process of CEA from cells, and that, in some cases, some epitopes may be masked by attachment of sialic acid residues.

Production kinetics and immunochemical properties of carcinoembryonic antigen and nonspecific cross-reacting antigen synthesized by various human tumor cell lines.

The production kinetics and immunochemical properties of carcinoembryonic antigen (CEA) and nonspecific cross-reacting antigen (NCA) in various human tumor cell lines were studied. By radioimmunoassay (RIA), five CEA-producing tumor cell lines tested--2 derived from colonic (M7609 and CCK-81), one from pancreatic (QGP-1) and 2 from lung (HLC-1 and KNS-62) carcinomas--were found to produce NCA simultaneously. The cellular contents of CEA and NCA and the amounts of both antigens released into the culture medium were highly variable among the cell lines. It was a distinct contrast that one cell line (CCK-81) released very large amounts of CEA and NCA into the medium while having the smallest amounts of both antigens in the cells, whereas the others contained much larger amounts of the antigens in the cells as compared with the amounts released into the medium. For most of the cell lines, the production of both CEA and NCA increased in the stationary phase of growth as compared with the exponential phase. The production kinetics of both CEA and NCA appeared to be parallel with each other in all the cell lines, though the amount ratio of CEA to NCA produced was variable. By means of a double immunodiffusion test with polyclonal antibodies, antigenic uniformity with no unique organ-specificity was confirmed for all the CEA preparations from spent media of the cell lines, though some differences in the sugar moiety of CEA were detected by RIA using monoclonal antibodies. No antigenic differences among NCA preparations were observed. Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), molecular heterogeneity was observed among CEA or NCA preparations isolated from cell lysates.

Production of immunoreactive calcitonin and some other tumor markers by established human carcinoma cell lines.

Out of seven human carcinoma cell lines (M7609, CCK-81, FCC-1, RPMI#4788, QGP-1, HLC-1, and KNS-62), 4 cell lines were found to produce immunoreactive calcitonin (ICT), a potential tumor marker for various malignancies. During a 7-day culture, 1.4 X 10(5) QGP-1, RPMI#4788, HLC-1, and KNS-62 cells secreted 7,000 pg, 500 pg, 400 pg, and 400 pg of ICT in the medium, respectively. The production of ICT by QGP-1 cells was increased by addition of pentagastrin or calcium gluconate. Three different components of ICT (peak I, molecular weight greater than 40,000; peak II, 14,000-18,000; peak III, 3,400) were detected by gel filtration of the QGP-1 spent medium. In a competitive inhibition-type radioimmunoassay of serial dilutions of each ICT component, peak III component showed very similar immunoreactivity to synthetic calcitonin. However, the other two components gave clearly different immunoreactivities from the peak III component and showed very similar immunoreactivities to each other. All the cell lines were further screened for synthesis of 7 other tumor markers, carcinoembryonic antigen, nonspecific cross-reacting antigen, CA19-9, tissue polypeptide antigen, alpha-fetoprotein, beta 2-microglobulin and ferritin. Every cell line produced 2 to 6 markers concomitantly, and various combinations of positive markers were found among the cell lines.

Identification and partial characterization of the unglycosylated peptide of carcinoembryonic antigen synthesized by human tumor cell lines in the presence of tunicamycin.

Unglycosylated peptide backbones of carcinoembryonic antigen (CEA) synthesized by human tumor cell lines in the presence of tunicamycin were identified and analyzed by SDS-polyacrylamide gel electrophoresis. Three tumor cell lines, QGP-1 (pancreas), FCC-1 (colon) and KNS-62 (lung) were found to produce CEA molecules of 180,000-190,000 mol. wts labeled with both [3H]leucine and [14C]glucosamine under conventional culture conditions. In contrast, in the presence of tunicamycin, the native CEA molecules disappeared, and a new component that was precipitated with anti-CEA antibodies and labeled only with [3H]leucine but not with [14C]glucosamine was identified in each cell line. Monoclonal antibodies each directed to different major antigenic determinants on the native CEA molecules also reacted with this unglycosylated peptide. The apparent mol. wts of the naked CEA peptides from QGP-1 and FCC-1 were equally about 78,000, whereas that from KNS-62 was somewhat larger than the other two, suggesting some differences in the peptide structure of the CEA molecules.

Coproduction of carcinoembryonic antigen and nonspecific cross-reacting antigen by a continuous cell line from a human pancreatic tumor.

A simultaneous production of nonspecific cross-reacting antigen (NCA) and carcinoembryonic antigen (CEA) by the same individual cells of an established human pancreatic cell line (QGP-1) was demonstrated by the immunoperoxidase method. Kinetics of cell proliferation and production of CEA and NCA were analyzed, and active synthesis of both antigens was found to be accompanied with the active proliferation of cultured cells. Both antigens in culture medium were purified by immunoadsorption and gel filtration. Immunochemical studies confirmed that CEA and NCA produced by the QGP-1 cells had properties identical to those of authentic CEA derived from metastatic colorectal carcinoma and to those of NCA from normal lungs, respectively.