Ultrastructural identification of uncoated caveolin-independent early endocytic vehicles

Using quantitative light microscopy and a modified immunoelectron microscopic technique, we have characterized the entry pathway of the cholera toxin binding subunit (CTB) in primary embryonic fibroblasts. CTB trafficking to the Golgi complex was identical in caveolin-1 null (Cav1 -/-) mouse embryonic fibroblasts (MEFs) and wild-type (WT) MEFs. CTB entry in the Cav1 -/- MEFs was predominantly clathrin and dynamin independent but relatively cholesterol dependent. Immunoelectron microscopy was used to quantify budded and surface-connected caveoloe and to identify noncaveolar endocytic vehicles. In WT MEFs a small fraction of the total Cav1-positive structures were shown to bud from the plasma membrane (2 % per minute), and budding increased upon okadaic acid or lactosyl ceramide treatment. However, the major carriers involved in initial entry of CTB were identified as uncoated tubular or ring-shaped structures. These carriers contained GPI-anchored proteins and fluid phase markers and represented the major vehicles mediating CTB uptake in both WT and caveolae-null cells.

Unsafe driving behaviour and four wheel drive vehicles: observational study

Objective To assess the level of compliance with the new law in the United Kingdom mandating penalties for rising a hand held mobile phone while driving, to compare compliance with this law with the one on the use of seat belts, and to compare compliance with these laws between drivers of four wheel drive vehicles and drivers of normal cars. Design Observational study with two phases-one within the grace period, the other starting one week after penalties were imposed on drivers using such telephones. Setting Three busy sites in London. Participants Drivers of 38 182 normal cars and 2944 four wheel drive vehicles. Main outcome measures Proportions of drivers seen to be using hand held mobile phones and not using seat belts. Results Drivers of four wheel drive vehicles were more likely than drivers of other cars to be seen using hand held mobile phones (8.2% v 2.0%) and not complying with the law on seat belts (19.5% v 15.0%). Levels of non-compliance with both laws were slightly higher in the penalty phase of observation, and breaking one law was associated with increased likelihood of breaking the other. Conclusions The level of non-compliance with the law on the use of hand held mobile phones by drivers in London is high, as is non-compliance with the law on seat belts. Drivers of four wheel drive vehicles were four times more likely than drivers of other cars to be seen using hand held mobile phones and slightly more likely not to comply with the law on seat belts.