Accumulation of rab4GTP in the cytoplasm and association with the peptidyl-prolyl isomerase pin1 during mitosis.

Transport through the endocytic pathway is inhibited during mitosis. The mechanism responsible for this inhibition is not understood. Rab4 might be one of the proteins involved as it regulates transport through early endosomes, is phosphorylated by p34(cdc2) kinase, and is translocated from early endosomes to the cytoplasm during mitosis. We investigated the perturbation of the rab4 GTPase cycle during mitosis. Newly synthesized rab4 was less efficiently targeted to membranes during mitosis. By subcellular fractionation of mitotic cells, we found a large increase of cytosolic rab4 in the active GTP-form, an increase not associated with the cytosolic rabGDP chaperone GDI. Instead, phosphorylated rab4 is in a complex with the peptidyl-prolyl isomerase Pin1 during mitosis, but not during interphase. Our results show that less efficient recruitment of rab4 to membranes and a bypass of the normal GDI-mediated retrieval of rab4GDP from early endosomes reduce the amount of rab4GTP on membranes during mitosis. We propose that phosphorylation of rab4 inhibits both the recruitment of rab4 effector proteins to early endosomes and the docking of rab4-containing transport vesicles. This mechanism might contribute to the inhibition of endocytic membrane transport during mitosis.

Vibrational spectrometry for the assessment of active substance in metoprolol tablets: a comparison between transmission and diffuse reflectance near-infrared spectrometry.

Near-infrared spectrometry (NIR) was used to quantify metroprolol succinate in controlled release tablets. Metoprolol tablets were made according to an experimental design using different strengths around a central strength of 47.5 mg per tablet. A comparison was made between NIR in the diffuse reflectance mode and the transmission mode. This showed that, although a narrower wavelength range was available in the transmission mode, predictions were much better for models based on transmission spectra than for models based on diffuse reflectance spectra. The main reason for this is that in the reflectance mode NIR spectrometry is very sensitive to the inhomogeneity of the material, while in the transmission mode this problem is less severe. This is due to the larger volume of the material scanned in the transmission mode compared to that in diffuse reflectance. Spectra were taken before and after the tablets were stored under humid conditions. This allowed the final calibration models to be made more robust towards variations in the amount of water in the tablet. Different batches of metoprolol pellets and microcrystalline cellulose were used during the production of the tablets. this resulted in models that were more robust towards possible batch-to-batch differences in the main constituents.