To pee or not to pee: no more the question in ARF?
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The formation and maturation of membrane carriers that transport cargo from the ER to the Golgi complex involves the sequential action of the coat protein complexes COPII and COPI. Recruitment of COPI to nascent carriers requires activation of ADP-ribosylation factors by a BrefeldinA-sensitive guanine nucleotide exchange factor. Using new antisera and a GFP-tagged protein, we demonstrate that the exchange factor GBF1 localized to both Golgi membranes and peripheral puncta, near but separate from ER exit sites. Live cell imaging revealed that GFP-GBF1 associates dynamically with both membranes through rapid exchange with a large cytosolic pool. Treatment with BrefeldinA dramatically altered this rapid exchange, causing accumulation of GBF1 on both Golgi and peripheral puncta before eventual redistribution to the ER in a microtubule-dependent manner. Measurement of diffusion coefficients and subcellular fractionation confirmed this shift in GBF1 from cytosolic to membrane bound. BrefeldinA-induced accumulation of GBF1 coincided with loss of COPI from peripheral puncta. Furthermore, recruitment of GBF1 to cargo-containing peripheral puncta coincided with recruitment of COPI, but not COPII. Strikingly, microinjection of anti-GBF1 antibodies specifically caused dissociation of COPI from membranes. These observations strongly suggest that GBF1 regulates COPI membrane recruitment in the early secretory pathway.
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Twenty male albino rabbits were studied. Four animals served as controls; the remaining 16 animals represented the treated group. All the treated animals were exposed to the same amount of energy delivered by the excimer laser (pulse rate: 20 Hz, fluence 250mJ/cm2; number of pulses: 6032; cumulative UV dose 1508 J/cm2) and were divided into eight groups of 2 animals each (four eyes). Samples of aqueous humor and lens were obtained at the following intervals: 5, 10, 20 and 40 min and 1, 2, 3 and 4 weeks after photorefractive keratectomy (PRK). The levels of reduced and oxidized glutathione, hydrogen peroxide, ascorbic acid and malondialdehyde were determined. Aqueous humor analyses, twenty min after PRK, showed no significant differences with pre-treatment values, while the observed variations in lens were constantly present over the entire follow-up period (one month). These findings suggest that the biochemical lens alterations induced by PRK may represent the earliest events relevant to cataractogenesis in the rabbit.
Nephrotoxicity is a dose limiting feature of cis-diamminedichloroplatinum (DDP) cancer chemotherapy. We have previously developed a model of DDP induced acute renal failure in the rat, which is characterized by non oliguric progressive azotemia. Protocols have been established in humans to prevent or diminish DDP associated renal alterations during the course of cancer chemotherapy. The present studies were designed to evaluate the effect of prior diuretic therapy, with furosemide, and enhanced solute diuresis, using dextrose and water as the sole source of drinking fluid, on DDP induced acute renal failure in the rat. As compared to water drinking controls neither the diuretic nor the enhancement of osmotic excretion effected DDP associated mortality. The courses of the acute renal failure observed in all three study groups were similar; however, there was a suggestion in the surviving animals that these maneuvers may have contributed to a more rapid return in renal function among rats not dying of DDP induced acute renal failure.
We report a case of rhabdomyolysis secondary to diabetic hyperosmolar nonketotic coma leading to acute renal failure. The patient had no overt muscle pain or swelling. Urinary myoglobin was negative. The patient was managed with continuous veno-venous hemodiafiltration (CVVHD) and his renal function subsequently recovered. The clinical features of five other reported cases of rhabdomyolysis caused by diabetic hyperosmolar nonketotic coma in the literature are reviewed.
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Cytogenetic changes are of pivotal prognostic significance in patients with de novo acute lymphoblastic leukemia (ALL). However, in some cases leukemic blasts can harbor gene lesions on a submicroscopic level without evidence of a corresponding abnormality by conventional cytogenetic studies. This can result in failure to recognize chromosomal abnormalities and inappropriate evaluation with respect to therapy assignments. To study the discrepancy in the detection of deletions of the short arm of chromosome 9 and deletions of tumor suppressor genes p15/p16/p14 on chromosome 9p21, we analyzed bone marrow samples from 92 patients with ALL both by cytogenetic analysis and by Southern blot. In 41 patients (45%), we found deletions of p15/p16/p14, which were homozygous in 27 and hemizygous in 14. Cytogenetic analysis demonstrated abnormalities of the short arm of chromosome 9 in the form of 9p- or del(9p21-22) in only 5 of the 41 patients (12%). Only 2 of 51 patients without gene deletions as detected by Southern blot revealed a 9p- abnormality, which was found only in a subpopulation of the cells. We demonstrate that deletions of the p15/p16/p14 genes on chromosome 9p21 are more frequent than indicated by cytogenetic analysis. Molecular techniques in addition to cytogenetic studies are necessary to detect otherwise-unrecognized genetic lesions of the short arm of chromosome 9.