Epidermal growth factor: structure, location, phosphorylation and regulation of its receptor

Epidermal growth factor (EGF) is a Mr 6045 polypeptide first characterized for its ability to stimulate mitogenesis in epidermal and epithelial cells. The first step in the action of the growth factor is its binding to specific, high affinity membrane receptors. These receptors have been studied in a number of tissues and cell culture lines. The level of EGF receptors is modulated by many agents. EGF down-regulates its receptor. In addition, the number of EGF receptors is decreased by other growth factors (platelet-derived growth factor; transforming growth factor), by many tumor promoters and by viral transformation. Several hormones also can regulate EGF binding in its target tissues.     

Changes in rat gastric proteases with age: effect of hydrocortisone.

Gel filtration on Sephadex G-75 of rat gastric mucosal preparation shows two peaks of proteolytic activity. These peaks differ at different ages of animals. The proteolytic activity of the gastric mucosa is spread over a wider range of pH in younger animals than older ones with a shift from higher pH towards lower pH values with increasing age. Hydrocortisone injection to nine day-old rats results in qualitative changes of gastric mucosal proteases, the proteolytic activity pattern of young treated animals being similar to that of untreated adult rat.     

Immunohistochemical localization of beta-LPH in the human hypothalamus.

In order to establish the presence of β-LPH and to clearly identify the nervous structures containing β-LPH in the human hypothalamus, an immunohistochemical localization of β-LPH was performed in this tissue. The immunohistochemical technique involved use of a specific antiserum to human β-LPH and the peroxidase-antiperoxidase complex. Immunostained neuronal cell bodies were observed in the arcuate nucleus whereas β-LPH-positive nervous fibers could be detected in a large area extending rostro-caudally from the anterior part of the paraventricular nucleus up to the mammillary bodies. Staining was completely abolished by previous immunoabsorption with β-LPH while β-endorphin and ovine γ-LPH_1–47 only partially prevented immunostaining. Although it cannot be excluded that the precursor 31K molecule, β-LPH_1–58 and/or β-endorphin are detected by the immunostaining, it is likely that β-LPH is at least partly responsible for the positive reaction.     

Open-heart surgery in elderly patients.

Between March 1975 and March 1980, 50 patients aged 70 to 78 years underwent open-heart surgery at the Montreal Heart Institute. Coronary bypass was performed in 23 patients, valve replacement in 16 and combined coronary and valve surgery in 11. There were four early deaths, all due to cardiac causes. Early postoperative complications occurred in 58% of the patients. There were seven late deaths, five in the valve replacement groups and two in the isolated coronary bypass group. The cumulative survival rate 5 years after surgery was estimated at 76%. While 82% of the survivors were in functional class III or IV before surgery, 90% were in class I or II when last seen, after an average postoperative follow-up of 3 years. Nonfatal late complications occurred in eight of the survivors, one of whom suffered a major hemorrhage due to anticoagulant therapy. Thus, open-heart operations can be offered to the elderly, with a low risk of operative death. The late clinical improvement, with a return to a normal lifestyle, justifies a surgical approach for patients in otherwise good general condition.     

Localization of 17 beta-hydroxysteroid dehydrogenase throughout gestation in human placenta

17 beta-Hydroxysteroid dehydrogenase (17 beta-HSD) is the enzyme responsible for the formation of all sex steroids in gonadal as well as extragonadal tissues. To obtain more information about the age-specific expression of 17 beta-HSD in the human placenta, we have localized this enzyme by immunocytochemistry at the light microscopic level at different periods of gestation. In the 7- and 9-week-old placenta, immunostaining was detected exclusively in the cytoplasm of the syncytiotrophoblast. Between the tenth and thirteenth weeks of gestation, immunolabeling was also observed in the cytoplasm of the cytotrophoblastic cells, suggesting that these cells could be transiently involved in the biosynthesis of sex steroids. Interestingly, between the fourteenth and twenty-fifth weeks of gestation, 17 beta-HSD was observed in both the cytoplasm and nucleus of the syncytiotrophoblast. The reaction product was much more intense in nuclei than in cytoplasm. During the last trimester of gestation, strong immunocytochemical staining was observed in all the nuclei of the syncytiotrophoblast, the cytoplasm being unstained. The meaning of this nuclear staining for 17 beta-HSD is still unclear and remains to be extensively investigated.     

IL-1 production by human thymic dendritic cells: studies on the interrelation with DC accessory function

Thymic dendritic cells (DC) have been proposed to play a critical role in the generation of immunocompetent T lymphocytes. Since IL-1 is widely considered to be an important second signal in T cell stimulation, we have studied the ability of isolated human thymic DC to produce IL-1. Using the EL4/CTLL conversion assay standardized with recombinant IL-1 beta (rIL-1 beta), we demonstrate that upon LPS-stimulation thymic DC produce small amounts of IL-1 as compared to peripheral blood monocytes (PBM). In contrast with PBM, DC IL-1 production is not influenced by indomethacin. IL-1 activity was detected in the supernatants of DC cultures from all thymuses tested, although quantitative variability was noted among individual thymic donors. The specificity of the active factor was confirmed by neutralization assays with anti-IL-1 beta mAb. On the other hand, we demonstrate that rIL-1 beta cannot substitute for nor amplify the accessory function of thymic DC and that anti-IL-1 beta mAb fails to block the DC accessory function. Thus we conclude that IL-1 beta might not be a major factor for the efficient DC accessory function toward mature thymocytes recently demonstrated in our laboratory. Of interest, IL-1 beta was also detected in the supernatants of DC-thymocyte cocultures in the absence of mitogenic factor, suggesting that thymocyte contacts can constitute a sufficient signal to induce DC to produce IL-1. These observations indicate that human thymic DC represent an intrathymic source of IL-1 whose role in thymocyte proliferation or maturation remains to be understood.     

Insights on the amino acid side-chain interactions of a synthetic T-cell determinant.

The effect of single amino acid substitutions at positions 18 and 20 on the T-cell determinant (TD) character of peptide p12-26 from lambda repressor protein and on its recognition by a monoclonal antibody was studied by means of 40 synthetic peptides of a length of 15 amino acids. ELISA competition experiments showed that the identity of amino acid at position 20 is very important for antibody recognition, whereas that of amino acid at position 18 is much less important. In contrast, both Leu 18 and Ala 20 are important residues in defining the TD character of peptide p12-26. The most tolerated replacements, ordered in increasing disrupting power are: Ala 20 by Cys, Ser or Gly and Leu 18 by Ile or Val. Any other amino acid replacement completely abolishes the TD capacity of peptide p12-26. The peptides used in this study were synthesized using a multiple solid-phase peptide synthesizer newly designed. Their purity was very high as shown by amino acid sequence experiments.     

High-performance liquid chromatographic analysis of 2',3'-dideoxyinosine in biological samples

A high-performance liquid chromatographic analysis for the anti-AIDS drug 2',3'-dideoxyinosine (ddI) in rat plasma and urine, with a limit of detection of 0.2 μg/ml and requiring a sample size of 100 μl is described. Diluted plasma or urine samples were extracted using a C₁₈ solid-phase extraction column. Retention of ddI on more polar solid-phase extraction columns was insufficient for sample clean-up. This method is useful for pharmacokinetic studies of ddI in small rodents.     

Translation initiation factor-dependent extracts from Saccharomyces cerevisiae

Translation initiation factor 4A- and 4E-dependent extracts were developed from Saccharomyces cerevisiae and used to study factor requirements for translation of individual mRNAs in vitro. Whereas all mRNAs tested required eIF-4A, mRNAs devoid of secondary structure in their 5' untranslated region did not require exogenous eIF-4E for translation. The latter included alfalfa mosaic virus RNA4, mRNA containing the untranslated region of tobacco mosaic virus RNA and mRNA containing part of the untranslated region of poliovirus RNA. Furthermore, initiation of translation on mRNAs containing part of the untranslated region of poliovirus RNA is most likely internal.