Biomedical subjects
Publications and source records attributed to S Jacobs.
The dexamethasone suppression test (DST) was given to 13 elderly grieving subjects having a major depressive disorder. Forty-six per cent had some predisposition for depression. All had a post-DST, 4:00 p.m. serum cortisol below 5 micrograms/dl, but these cortisols correlated with Hamilton Depression scores (r = .59, p less than .05).
Insulin receptors from rat hepatoma cells (Fao) and human placenta were partially purified by detergent solubilization and lectin purification. The insulin receptor preparations were subjected to sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis under reducing or nonreducing conditions. The proteins were transferred to nitrocellulose paper and the electrophoretic blots were treated with human anti-receptor autoantibodies, rabbit antibody to purified insulin receptor, or a monoclonal antibody to human insulin receptor. The nitrocellulose paper was then treated with 125I protein A or 125I second antibody followed by autoradiography. The rabbit polyclonal antiserum and one of the human autoantibodies recognized both the alpha (Mr = 135,000) and beta (Mr = 95,000) subunits after transfer from a SDS gel to nitrocellulose paper. On transfers from nonreduced gels, several high-molecular species were labeled ranging from Mr = 200,000 to Mr = 330,000. Similar high-molecular bands of the receptor were seen if highly purified human placental receptor, as well as partially purified receptor from rat or human origin, were used. As little as 0.1-0.5 microgram of pure receptor could be detected by this technique. Treatment of the receptor with neuraminidase (50 mU/ml) before gel electrophoresis resulted in a 50% increase in intensity of intact receptor and about a 70% increase in the labeling of the alpha-subunit of the receptor, but no change in labeling of the beta-subunit. The monoclonal antibody used, as well as two other human autoantibodies, did not recognize the receptor after transfer to nitrocellulose paper.(ABSTRACT TRUNCATED AT 250 WORDS)
Explore the source record for details and available documents.
Explore the source record for details and available documents.
A patient with suspected splenic trauma is reported in whom, because of an anatomic variant, both radionuclide and sonographic examinations were misleading and led to the erroneous diagnosis of splenic laceration. Confusion arose because of an overlying left lobe of liver and this was clarified by biliary scintigraphy.
Phosphorylation of the somatomedin-C receptor was investigated both in intact IM-9 cells and in IM-9 cells that had been solubilized with Triton X-100. Intact IM-9 cells were incubated with [32P]H3PO4 for 1 h and for an additional 5 min in the absence or presence of insulin or somatomedin-C. The cells were then solubilized and subjected to wheat germ agglutinin Sepharose chromatography. The extent of phosphorylation of insulin and somatomedin-C receptors was assessed by immunoprecipitating the wheat germ agglutinin Sepharose eluates with monoclonal antibodies specific for each receptor and analyzing the immunoprecipitates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The beta-subunits of both receptors were phosphorylated in the absence of hormone, and the extent of phosphorylation of each receptor was enhanced by both hormones. However, each hormone was more potent than the other in enhancing phosphorylation of its own receptor. The beta-subunit of the somatomedin-C receptor was also phosphorylated when solubilized IM-9 cells that had been purified on wheat germ agglutinin Sepharose were incubated with [gamma-32P]ATP. In this soluble preparation, phosphorylation occurred on tyrosyl residues and was enhanced by concentrations of somatomedin-C in the range of 2.5 to 250 ng/ml, which is consistent with its receptor affinity. Tyrosyl phosphorylation of the somatomedin-C receptor also occurred when highly purified receptor, prepared by wheat germ agglutinin Sepharose affinity chromatography followed by immunoprecipitation, was incubated with [gamma-32P]ATP. This indicates that the responsible tyrosyl kinase activity is intrinsic to the receptor or tightly associated with it.
Three monoclonal antibodies, designated alpha IR-1, alpha IR-2, and alpha IR-3, were prepared by fusing FO myeloma cells with spleen cells from a mouse immunized with a partially purified preparation of insulin receptors from human placenta. These antibodies were characterized by their ability to immunoprecipitate solubilized receptors labeled with 125I-insulin or 125I-somatomedin-C in the presence or absence of various concentrations of unlabeled insulin or somatomedin-C. alpha IR-1 preferentially immunoprecipitates insulin receptors and also less effectively immunoprecipitates somatomedin-C receptors, while alpha IR-2 and alph IR-3 preferentially immunoprecipitate somatomedin-C receptors, but may also weakly immunoprecipitate insulin receptors. These three monoclonal antibodies, as well as A410, a rabbit polyclonal antibody, were used to immunoprecipitate insulin and somatomedin-C receptors from solubilized human lymphoid (IM-9) cells and human placenta membranes that had been 125I-labeled with lactoperoxidase. Analysis of the immunoprecipitates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicates that both receptors are composed of alpha and beta subunits. The beta subunit of the insulin receptor (immunoprecipitated by alpha IR-1 and A410) has a slightly more rapid mobility than the corresponding subunit of the somatomedin-C receptor (immunoprecipitated by alpha IR-2 and alpha IR-3). Interestingly, the alpha subunit of the placenta somatomedin-C receptor has a slightly faster mobility than its counterpart from IM-9 cells. Immunoprecipitation of receptor that had been reduced and denatured to generate isolated subunits indicates that alpha IR-2 and alpha IR-3 interact with the alpha subunit of the somatomedin-C receptor while A410 interacts with both subunits of the insulin receptor. alpha IR-1 failed to react with reduced and denatured receptors.
Incubation of rat liver plasma membranes with insulin enhances the production of small molecular weight substances which regulate the activity of liver acetyl CoA carboxylase. While low concentrations of insulin cause the release of a carboxylase stimulator from membranes, concentrations greater than 10(-9) M generate less stimulating activity. This biphasic concentration curve for insulin can be resolved by differential alcohol extraction into two fractions which have antagonistic activity. The production of both substances is enhanced by insulin. Chemical and chromatographic evidence suggest that these substances are identical to the previously described "mediators" which regulate both pyruvate dehydrogenase and adenylate cyclase activities.
The effect of phorbol esters on the extent of phosphorylation of receptors for insulin and somatomedin C (insulin-like growth factor I) was studied in intact IM-9 cells that were labeled by incubation with H332PO4. The tumor-promoting phorbol esters phorbol tetradecanoate acetate (TPA) and phorbol dibutyrate, but not the inactive 4 alpha-phorbol, enhanced phosphorylation of the beta subunit of both receptors approximately 4-fold; 70 nM TPA maximally stimulated phosphorylation of both receptors, whereas concentrations less than or equal to 0.7 nM had no observable effect. Insulin also enhanced the phosphorylation of the beta subunit of the insulin receptor, and its effects appeared to be additive to those of TPA. Peptide maps indicated that at least some of the residues phosphorylated by these two agents are distinct. These results suggest a possible role of protein kinase C in regulating insulin and somatomedin C receptors.
Cultured human lymphoid (IM-9) cells were labeled with [(35)S]methionine in the presence and absence of monensin, a carboxylic ionophore that inhibits post-translational protein maturation. Labeled receptors for insulin and somatomedin C were immunoprecipitated with antibodies specific for each receptor. Monensin inhibits the biosynthesis of mature alpha and beta subunits of both receptors and leads to the accumulation of immunoreactive polypeptides with molecular weights of 180,000. These 180,000 molecular weight polypeptides exist as disulfide-linked dimers and may be biosynthetic precursors of both alpha and beta subunits. In the presence of monensin, small amounts of immunoreactive polypeptides with molecular weights 115,000 and 89,000 also are produced. These may be abnormally processed forms of the alpha and beta subunits lacking residues normally added during terminal glycosylation. In cells treated with monensin, the polypeptides of molecular weights 180,000 and 115,000 can be affinity-labeled with (125)I-labeled insulin. These labeled polypeptides are immunoprecipitated by antibodies specific for insulin receptors but not by antibodies specific for somatomedin-C receptors. This indicates that the putative precursors for insulin and somatomedin-C receptors are distinct polypeptides, although they have similar molecular weights and similar modes of processing. A possible structural relationship between the precursors for these receptors and the type II insulin-like growth factor receptor is discussed.
Adrenocortical function was tested in 12 patients following a single lumbar extradural injection of methylprednisolone acetate ('Depo-Medrol') 80 mg as treatment for chronic sciatica. There was no absorption of the corticosteroid into the systemic circulation, but marked suppression of plasma cortisol levels was documented for up to 3 weeks following the injection and the capacity of the adrenal cortex to secrete cortisol in response to synthetic adrenocorticotrophin (ACTH) was diminished. These results suggest that the dose and frequency of extradural steroid administration should be kept to a minimum to prevent suppression of the hypothalamic/pituitary/adrenocortical axis and that patients thus treated should be considered candidates for steroid cover during surgery and other stressful procedures.
Explore the source record for details and available documents.
The insulin receptor has been purified by affinity chromatography and studied by affinity-labelling techniques. It appears to be a disulphide-linked heterotetramer, (alpha beta)2, composed of two copies of a 135,000 Mr subunit (alpha), and two copies of a 90,000 Mr subunit (beta). Beta is readily proteolysed to generate a 45,000 Mr fragment (beta 1). Alpha, beta and beta 1 all contain sialic acid and are, therefore, probably all exposed on the external surface of the membrane. Although alpha is predominantly labelled in affinity-labelling studies, beta and beta 1 can also be labelled. Therefore, alpha, beta and beta 1 are all in proximity to the insulin-binding site and may contain part of the binding site. Antibodies have been prepared against the intact, purified receptor and against the isolated alpha subunit. Both antibodies directly interact with the insulin receptor as indicated by their ability to immunoprecipitate the receptor. Neither antibody, however, directly competes with insulin binding. Therefore, they are probably directed against regions of the receptor distinct from the insulin-binding site. In spite of this, these antibodies have a wide range of insulin-like activities.
Explore the source record for details and available documents.
Recent evidence suggests that certain actions of insulin may be mediated by the selective generation of chemically undefined intracellular substances. Incubation of rat liver particulate fraction with low concentrations of insulin enhances the release into the supernatant of a substance that stimulates mitochondrial pyruvate dehydrogenase. Higher concentrations of insulin release less stimulating activity. It is possible to resolve activities that stimulate and inhibit pyruvate dehydrogenase by differential ethanol extraction of the supernatant solutions. The elaboration of both factors is dependent upon the presence of insulin in a dose-dependent manner. Moreover, fractions that contain the pyruvate dehydrogenase-inhibiting activity also inhibit adipocyte basal and hormonally stimulated adenylate cyclase. The production of this adenylate cyclase inhibitory activity is also stimulated by insulin. Cyclase inhibition is virtually abolished when the nonhydrolyzable ATP analog, 5'-adenylyl imidodiphosphate, is included in the assay. These results indicate that the bimodal effects of insulin on certain functions may be ascribed to the generation of at least two distinct chemical substances that show opposing activities, which may operate by regulating phosphorylation reactions.
Explore the source record for details and available documents.
To identify constituents of grain dust responsible for grain handlers' respiratory symptoms, 11 volunteer grain elevator workers underwent inhalation provocation tests with extracts of durum wheat, durum wheat airborne dust, and grain insects and mites. Factors that might influence the host response to the challenge were assessed. Five of the 11 subjects showed a greater than 20 percent decrement in FEV1 after inhalation of durum wheat extracts (airways reaction). The bronchial reactions were immediate in 1/5 and late in 4/5. These airway reactions were blocked by sodium cromoglycate. Only one subject showed airway reaction to durum wheat dust extract, and none reacted to mites or insect extracts. There was no change in DLCO, temperature, or in total C3 complement blood levels, nor was there evidence of activation of complement by the alternate or classic pathway. The airway response was not always related to the atopic status of the individual. The reaction is more likely to occur in subjects with preexisting airway obstruction and nonspecific bronchial hyperreactivity. Durum wheat has been identified as an inducer of occupational asthma in grain handlers. This reaction is probably due to specific or nonspecific mediator release. There was no apparent parenchymal or systemic reaction or complement activation detectable in the subjects' sera after inhalation of durum wheat extracts.