Involvement of ATP-dependent Pseudomonas Exotoxin Translocation from a Late Recycling Compartment in Lymphocyte Intoxication Procedure

Pseudomonas exotoxin (PE) is a cytotoxin which, after endocytosis, is delivered to the cytosol where it inactivates protein synthesis. Using diaminobenzidine cytochemistry, we found over 94% of internalized PE in transferrin (Tf) -positive endosomes of lymphocytes. When PE translocation was examined in a cell-free assay using purified endocytic vesicles, more than 40% of endosomal 125I-labeled PE was transported after 2 h at 37°C, whereas a toxin inactivated by point mutation in its translocation domain was not translocated. Sorting of endosomes did not allow cell-free PE translocation, whereas active PE transmembrane transport was observed after > 10 min of endocytosis when PE and fluorescent-Tf were localized by confocal immunofluorescence microscopy within a rab5-positive and rab4- and rab7-negative recycling compartment in the pericentriolar region of the cell. Accordingly, when PE delivery to this structure was inhibited using a 20°C endocytosis temperature, subsequent translocation from purified endosomes was impaired. Translocation was also inhibited when endosomes were obtained from cells labeled with PE in the presence of brefeldin A, which caused fusion of translocation-competent recycling endosomes with translocation-incompetent sorting elements. No PE processing was observed in lymphocyte endosomes, the full-sized toxin was translocated and recovered in an enzymatically active form. ATP hydrolysis was found to directly provide the energy required for PE translocation. Inhibitors of endosome acidification (weak bases, protonophores, or bafilomycin A1) when added to the assay did not significantly affect 125I-labeled PE translocation, demonstrating that this transport is independent of the endosome-cytosol pH gradient. Nevertheless, when 125I-labeled PE endocytosis was performed in the presence of one of these molecules, translocation from endosomes was strongly inhibited, indicating that exposure to acidic pH is a prerequisite for PE membrane traversal. When applied during endocytosis, treatments that protect cells against PE intoxication (low temperatures, inhibitors of endosome acidification, and brefeldin A) impaired 125I-labeled PE translocation from purified endosomes. We conclude that PE translocation from a late receptor recycling compartment is implicated in the lymphocyte intoxication procedure.

Non-alcoholic fatty liver disease associated with hypobetalipoproteinemia: report of three cases and a novel mutation in APOB gene

Background: Non-alcoholic fatty liver disease, the leading cause of chronic liver disease in children, is defined by hepatic fat infiltration >5% of hepatocytes, in the absence of excessive alcohol intake, evidence of viral, autoimmune or drug-induced liver disease. Conditions like rare genetic disorders must be considered in the differential diagnosis. Case Report: Two male brothers, and a non-related girl, all overweight, had liver steatosis. One of the brothers and the girl had elevated transaminases; all three presented with low total cholesterol, low density lipoproteins and very low density lipoproteins cholesterol levels, hypotriglyceridemia and low apolipoprotein B. A liver biopsy performed in the brother with citolysis confirmed steatohepatitis and the molecular study of apolipoprotein B gene showed a novel homozygous mutation (c.9353dup p.Asn3118Lysfs17). Patients with cytolysis lost weight, however liver steatosis persists. Conclusion: Fatty liver disease might be a consequence of hypobetalipoproteinemia. Evidence is scarce due to low number of reported cases.

Analytical study of the legal and operational aspects of the Minnesota law entitled "Chemical test for intoxication" - M.S.A. Sec. 169.123. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Data pertinent to a review of minimum wage rates established under the Fair labor standards act for newly covered employment in the alcoholic beverage and industrial alcohol industry in Puerto Rico /

"WH65-69."

Assessment of clinical procedures to evaluate liver intoxication in fish /

Mode of access: Internet.

Comparison of recovered alcoholic and non-alcoholic communicator in alcohol education. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

State alcohol related fatality rates.

Mode of access: Internet.

On the use and abuse of alcoholic liquors : in health and disease /

Mode of access: Internet.

Rehabilitation of the alcoholic; a report from the Study Group on Rehabilitation of the Alcoholic and Public Offender.

Report presented at the Sixth Institute on Rehabilitation Services, held May 20-22, 1968, at University Park, Pa.

Studies in animal poisoning and intoxication-USSR.

Translations of two articles from Farmakologiyz i Toksikologiya, no.5, Sept.-Oct. 1968, published in Moscow.

Alcoholism Treatment Licensing Act and alcoholism and intoxication treatment programs : (77 Illinois Administrative Code 205 [i.e. 200]).

"These rules for the licensing of alcoholism treatment programs and facilities are numbered and labelled to follow the codification system for rules of all state agencies, which was developed in 1984"--Page 2.

Alcoholism. The subcommittee to study the problems of effective implementation of a comprehensive program of care and treatment for alcoholic individuals and public inebriates of the state of Illinois.

Cover title.