Increased cell surface EGF receptor expression during the butyrate-induced differentiation of human HCT-116 colon tumor cell clones

Several clonal sublines of HCT-116 human colon adenocarcinoma cells were isolated and characterized on the basis of their growth characteristics, intrinsic enterocyte-like differentiation (as assessed by alkaline phosphatase and lactase activities), and responses to butyrate, an inducer of colon tumor cell maturation. The HCT-116 sublines were found to be heterogeneous and several phenotypically distinct clones were identified. Further characterization of these clones indicated that the effects of butyrate on cell growth, alkaline phosphatase activity, and lactase activity were distinct and separable. The growth of all of the clones were inhibited by butyrate (IC50 values varied from 0.44 to 1.5 mM), but the effects of this agent on alkaline phosphatase and lactase activities varied widely. In several sublines butyrate had no effect on either enzyme while in others one or both activities were induced. Additionally, the binding of 125I-epidermal growth factor (EGF) to cell surface receptors was found to be proportional to the expression of lactase activity in the cell. The D3 clone and other sublines with intrinsic lactase activities greater than 100 nmol/mg/min expressed a class of high-affinity EGF receptors (e.g., D3 cells had 3.48 X 10(4) EGF receptors/cell with a kd of 0.61 nM). Other clones with less lactase activity had undetectable levels of 125I-EGF binding. In clones which exhibited greater than twofold increases in lactase activity in response to butyrate, the expression of a large number of low-affinity EGF receptors was also induced. In one such clone, the P1 subline, lactase activity was increased from 70 nmol/mg/min to 230 nmol/mg/min after 96 h in 2 mM butyrate, and the expression of EGF receptors was increased from undetectable levels to 1.18 X 10(5) EGF receptors/cell (kd of 3.2 nM). Northern blot analysis indicated that the increased 125I-EGF binding after butyrate treatment may have been due, in part, to a greater than twofold accumulation of EGF receptor mRNA. In addition, the expression of the messages for transforming growth factor alpha (TGF-alpha) and transforming growth factor beta (TGF-beta) was examined in butyrate-treated cells. While TGF-alpha mRNA levels were found to correlate with EGF receptor message levels in the HCT-116 clones, TGF-beta mRNA expression was not found to correlate with the butyrate-induced growth inhibition or with increases in EGF receptor expression, alkaline phosphatase activity, or lactase activity in these cells.     

Human papillomavirus infection of the cervix: the atypical condyloma

We report on 162 cases of human papillomavirus (HPV) infection of the cervix seen in a two-year period in which the cell sample showed such marked atypia that errors of interpretation could easily have been made. These atypical condylomata are difficult to diagnose cytologically as well as histologically because they mimic dysplasia or carcinoma in situ and, on smears, even invasive squamous carcinoma. HPV particles associated with fibrillar material were found within nuclei of these lesions; their nature was further proved by the immunoperoxidase test. This new form of HPV infection of the cervix showed a 9.1% rate of progression to more advanced cervical lesions. The cytologic finding of atypical condylomata is an indication for colposcopy, confirmative biopsy and appropriate treatment.     

On the molecular mechanism of interaction between the neurophysins and oxytocin and vasopressin

Detailed mechanisms are presented at the molecular level for the binding of oxytocin and of vasopressin to their carrier proteins neurophysin (NP) I and neurophysin II. The amino acid sequence of both these is known together with the pattern of disulphide bound formation for the latter. It is suggested that the peptide hormone fits snugly into a deep cavity in the protein carrier, so that the complex forms a globular, water-extruding mass. Features of the mode of interaction determined by experiment [such as the binding of the terminal amino group of the hormone to a carboxyl group of NP, the close binding of tyr and phe of the hormone to a lipophilic region of NP and the close relation of tyr (2) of the hormone with tyr (49) of NP]are built into the model. The differences between NPI and NPII are related to differences between oxytocin (ile at 3) and vasopressin (phe at 3). Finally a number of specific predictions are made that are testable by experiment concerning the X-ray structure of the NP-hormone complexes and the ease and result of chemical modification of specific residues.     

Purification,characterization and biological activity of three forms of ferredoxin from the sulfate-reducing bacterium Desulfovibrio gigas

Three forms of ferredoxin FdI, FdI′, and FdII have been isolated from Desulfovibrio gigas, a sulfate reducer. They are separated by a combination of DEAE-cellulose and gel filtration chromatographic procedures. FdI and FdI′ present a slight difference in isoelectric point which enables the separation of the two forms over DEAE-cellulose, while FdII is easily separated from the two other forms by gel filtration. The three forms have the same amino acid composition and are isolated in different aggregation states. Molecular weight determinations by gel filtration gave values of 18 000 for FdI and FdI′ and 24 000 for FdII, whereas a value of 6000 is determined when dissociation is accomplished with sodium dodecyl sulfate. The electronic spectra are different and their ultraviolet-visible absorbance rations are 0.77, 0.87 and 0.68 respectively for FdI, FdI′ and FdII. Despite these differences, the physiological activities of the three forms are similar as far as the reduction of sulfite by molecular hydrogen is concerned.     

Mycologie medicale dans l'ouest de la France: Contribution du service de Parasitologie de l'Universite de NANTES (1963–1973)

Résumé Le service de Parasitologie de l'Université de NANTES s'est intéressé aux problèmes de Mycologie hospitalière, d'épidémiologie des mycoses ainsi que d'expérimentation sur les affinités fongiques, à l'échelon régional, et de 1963 à 1973.La contribution clinique a porté essentiellement sur les levuroses étudiées soit au niveau des cavités buccales irradiées, soit chez les malades des services de Réanimation où l'interprétation du rôle pathogène de Candida parapsilosis nécessite des épreuves immunologiques.Nous avons décrit une forme de Blastomycose chéloïdienne à Aureobasidium pullulans.Nos études morphologiques ont surtout porté sur l'observation au microscope électronique à balayage des ultrasculptures présentées par les champignons kératinophiles, ce qui nous a permis de distinguer les différentes espèces du complexe Microsporum gypseum. Il ne semble pourtant pas que cette technique d'observation permette de résoudre tous les problèmes. La conservation des Dermatophytes en eaudistillée par la méthode de Castellani s'est révélée particulièrement remarquable: une souche de M. gypseum a été récupérée après 6 ans de conservation par ce procédé.Notons, en matière d'épidémiologie la prédominance de Trichophyton rubrum sur T. mentagrophytes, survenue récemment: la recrudescence passagère d'Epidermophyton floccosum; la présence d'Arthroderma simii sur une plage. Une étude des champignons kératinophiles telluriques du massif armoricain a porté sur près de 3000 échantillons de terre, ce qui a permis de retrouver, assez rarement d'ailleurs, Arthroderma benhamiae.Nos expérimentations ont porté sur les réponses sérologiques et histologiques de lapins à une imprégnation aspergillaire par instillation intra-trachéale: les résultats obtenus ont été comparés aux examens sérologiques obtenus chez l'homme atteint d'aspergillose. Enfin, nous avons remarqué que le contact levures-cellules sarcomateuses opéré in vitro permet aux levures d'acquérir un pouvoir immunisant antitumoral par emprunt antigénique. La création d'un service de Parasitologie, Mycologie et Immunologie Parasitaire à l'Unité d'Enseignement et de Recherche des Techniques Médicales de NANTES nous a permis non seulement d'aborder les problèmes du diagnostic biologique des mycoses humaines, mais aussi d'entreprendre des enquêtes épidémiologiques dans la région nantaise ainsi que des expériences sur les affinités fongiques.

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