RESUMO
We have recently reported a new oxidant- and calcium-inducible mRNA, adapt78, from hamster HA-1 cells. The adapt78 mRNA is induced in HA-1 cells under conditions where a protective adaptive response is observed and contains a translatable open reading frame whose protein product shows strong homology to a human sequence. Computer analysis of the predicted Adapt78 protein sequence also revealed a stretch of amino acids homologous to a portion of the glucose-regulated protein78 (Grp78). Based on this homology, we tested the hypothesis that adapt78 may be a new member of the grp gene family. Toward this, we assessed the modulation of adapt78 mRNA by stress agents known to induce grp78. In HA-1 cells, adapt78 mRNA was induced by the calcium ionophore A23187, 2-deoxyglucose, brefeldin A, tunicamycin, thapsigargin, and cyclopiazonic acid, with thapsigargin being the most potent inducer (7.3-fold). As expected, grp78 mRNA was also induced by these agents in our model system. In contrast, heat shock treatment produced little if any modulation of either grp78 or adapt78. Differences were also observed, as adapt78 mRNA but not grp78 mRNA was induced by 160 microM hydrogen peroxide, and adapt78 demonstrated earlier induction kinetics for certain agents compared with grp78. adapt78 mRNA was also found to be induced in several different human cell lines. A23187 had the strongest effect on adapt78 mRNA levels in human cells, inducing greater than 20-fold in all human cell cultures tested. Furthermore, in vitro transcription translation of human adapt78 cDNA produced an Adapt78 protein product. We conclude that adapt78 may be a new member of the grp family of genes and may represent an early response grp that complements the actions of grp78 and grp94.
Assuntos
Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico , Proteínas de Membrana/genética , Família Multigênica , RNA Mensageiro/genética , Sequência de Aminoácidos , Animais , Calcimicina/farmacologia , ATPases Transportadoras de Cálcio/antagonistas & inibidores , Proteínas de Transporte/genética , Linhagem Celular , Cricetinae , Desoxiglucose/farmacologia , Retículo Endoplasmático/enzimologia , Chaperona BiP do Retículo Endoplasmático , Inibidores Enzimáticos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Temperatura Alta , Humanos , Ionóforos/farmacologia , Chaperonas Moleculares/genética , Dados de Sequência Molecular , RNA Mensageiro/metabolismo , Homologia de Sequência de AminoácidosRESUMO
Traditional, less sensitive simple immunoassays ('Placentex' and 'Pregnosis') were performed concurrently with newer, more sensitive, but simple immunoassays ('Sensitex' and a capillary tube pregnancy test) and radioimmunoassays for beta-HCG from two commercial sources (Roche Diagnostics and Monitor Science Corporation). Generally these various assays agreed, but of particular importance for the early screening of urinary HCG (prior to the first missed but expected period) was that the commercial half-unit tube test ('Sensi-tex') gave early positive results comparable to the half-unit research capillary tube test previously reported. These sensitive simple immunoassays (SSIA) may improve the management of patients very early in pregnancy and following termination of pregnancy.
Assuntos
Gonadotropina Coriônica/urina , Imunoensaio/métodos , Feminino , Humanos , Gravidez , RadioimunoensaioRESUMO
Data are presented to show that human chorionic gonadotropin (hCG) in urine can be detected early in pregnancy by simple immunoassays performed prior to or around the time of the expected but missed menstrual period. Differences between the use of simple immunoassays with urine and radioimmunoassays and radioreceptor assays with serum are discussed.
Assuntos
Gonadotropina Coriônica/urina , Feminino , Humanos , Imunoensaio , Gravidez , Testes Imunológicos de Gravidez , Radioimunoensaio , Ensaio Radioligante , Fatores de TempoRESUMO
Further evaluation of the nonrefrigerated capillary tube pregnancy test is presented. A 127-mm by 2-mm glass capillary tube containing lyophilized antibody to human chorionic gonadotropin and latex indicator particles is used. Urine is drawn into the tube by capillary action; then, the tube is tilted to allow the urine to dissolve the antibody and to suspend the latex particles. The test results are read after a one-hour incubation period. Test results were evaluated against histopathologic and clinical data. Special efforts were made to stress the false-positive rate of the test by tripling the number of negative cases and doubling the sample size. A 99.82% agreement, a 0.18% false-negative rate and a 0% false-positive rate were obtained. In 52 confirmed pregnancies, the more sensitive capillary test detected human chorionic gonadotropin, whereas the commercial tests did not. Human chorionic gonadotropin was also detected one week earlier than reported in the first series.
