Expression cloning from a cDNA library prepared from a mutant CHO cell line with Golgi-specific resistance to Brefeldin A (BFA) identified a novel 206-kD protein with a Sec7 domain termed GBF1 for Golgi BFA resistance factor 1. Overexpression of GBF1 allowed transfected cells to maintain normal Golgi morphology and grow in the presence of BFA. Golgi- enriched membrane fractions from such transfected cells displayed normal levels of ADP ribosylation factors (ARFs) activation and coat protein recruitment that were, however, BFA resistant. Hexahistidine-tagged–GBF1 exhibited BFA-resistant guanine nucleotide exchange activity that appears specific towards ARF5 at physiological Mg2+concentration. Characterization of cDNAs recovered from the mutant and wild-type parental lines established that transcripts in these cells had identical sequence and, therefore, that GBF1 was naturally BFA resistant. GBF1 was primarily cytosolic but a significant pool colocalized to a perinuclear structure with the β-subunit of COPI. Immunogold labeling showed highest density of GBF1 over Golgi cisternae and significant labeling over pleiomorphic smooth vesiculotubular structures. The BFA-resistant nature of GBF1 suggests involvement in retrograde traffic.
Household humidification is widely practiced to combat dry indoor air. While the benefits of household humidification are widely perceived, its implications to the indoor air have not been critically appraised. In particular, ultrasonic humidifiers are known to generate fine particulate matter (PM). In this study, we first conducted laboratory experiments to investigate the size, quantity, and chemical composition of PM generated by an ultrasonic humidifier. The mass of PM generated showed a correlation with the total alkalinity of charge water, suggesting that CaCO 3 is likely making a major contribution to PM. Ion chromatography analysis revealed a large amount of SO 4 2− in PM, representing a previously unrecognized indoor source. Preliminary results of organic compounds being present in humidifier PM are also presented. A whole-house experiment was further conducted at an actual residential house, with five low-cost sensors (AirBeam) monitoring PM in real time. Operation of a single ultrasonic humidifier resulted in PM 2.5 concentrations up to hundreds of μg m −3 , and its influence extended across the entire household. The transport and loss of PM 2.5 depended on the rate of air circulation and ventilation. This study emphasizes the need to further investigate the impact of humidifier operation, both on human health and on the indoor atmospheric chemistry, for example, partitioning of acidic and basic compounds.
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