Primary cilia contain specific receptors and channel proteins that sense the extracellular milieu. Defective ciliary function causes ciliopathies such as autosomal dominant polycystic kidney disease (ADPKD). However little is known about how large ciliary transmembrane proteins traffic to the cilia. Polycystin-1 (PC1) and-2 (PC2), the two ADPKD gene products, are large transmembrane proteins that colocalize to cilia where they act to control proper tubular diameter. Here we describe that PC1 and PC2 must interact and form a complex to reach the trans-Golgi network (TGN) for subsequent ciliary targeting. PC1 must also be proteolytically cleaved at a GPS site for this to occur. Using yeast two-hybrid screening coupled with a candidate approach, we identify a Rabep1/GGA1/Arl3-dependent ciliary targeting mechanism, whereby Rabep1 couples the polycystin complex to a GGA1/Arl3-based ciliary trafficking module at the TGN. This study provides novel insights into the ciliary trafficking mechanism of membrane proteins.

Autosomal recessive polycystic kidney disease (OMIM 263200) is a serious condition of the kidney and liver caused by mutations in a single gene, PKHD1. This gene encodes Fibrocystin/Polyductin (FPC, PD1), a large protein shown by in vitro studies to undergo Notch-like processing. Its cytoplasmic tail, reported to include a ciliary targeting sequence, a nuclear localization signal and a polycystin-2 binding domain, is thought to traffic to the nucleus after cleavage. We now report a novel mouse line with a triple HA-epitope “knocked-in” to the C-terminus along with lox P sites flanking exon 67, which encodes most of the C-terminus (Pkhd1Flox67HA). The triple HA-epitope has no functional effect as assayed by phenotype and allows in vivo tracking of Fibrocystin. We used the HA tag to identify previously predicted Fibrocystin cleavage products in tissue. In addition, we found that Polycystin-2 fails to co-precipitate with Fibrocystin in kidney samples. Immunofluorescence studies with anti-HA antibodies demonstrate that Fibrocystin is primarily present in a sub-apical location in kidney, biliary duct and pancreatic ducts, partially overlapping with the Golgi. In contrast to previous studies, the endogenous protein in the primary cilia was not detectable in mouse tissues. After Cre-mediated deletion, homozygous Pkhd1Δ67 mice are completely normal. Thus, Pkhd1Flox67HA is a valid model to track Pkhd1-derived products containing the C-terminus. Significantly, exon 67 containing the nuclear localization signal and the polycystin-2 binding domain is not essential for Fibrocystin function in our model.

This study attempts to quantify the relative contributions of vegetation greening in China due to climatic and human influences from multiple observational datasets. Satellite measured vegetation greenness, Normalized Difference Vegetation Index (NDVI), and relevant climate, land cover, and socioeconomic data since 1982 are analyzed using a multiple linear regression (MLR) method. A statistically significant positive trend of average growing-season (April-October) NDVI is found over more than 34% of the vegetated areas, mainly in North China, while significant decreases in NDVI are only seen in less than 5% of the areas. The relationships between vegetation and climate (temperature, precipitation, and radiation) vary by geographical location and vegetation type. We estimate the NDVI changes in association with the non-climatic effects by removing the climatic effects from the original NDVI time series using the MLR analysis. Our results indicate that land use change is the dominant factor driving the long-term changes in vegetation greenness. The significant greening in North China is due to the increase in crops, grasslands, and forests. The socioeconomic datasets provide consistent and supportive results for the non-climatic effects at the provincial level that afforestation and reduced fire events generally have a major contribution. This study provides a basis for quantifying the non-climatic effects due to possible human influences on the vegetation greening in China.

In recent years, paper-based point-of-care testing (POCT) has been widely used in medical diagnostics, food safety and environmental monitoring. However, a high-cost, time-consuming and equipment-dependent sample pretreatment technique is generally required for raw sample processing, which are impractical for low-resource and disease-endemic areas. Therefore, there is an escalating demand for a cost-effective, simple and portable pretreatment technique, to be coupled with the commonly used paper-based assay (e.g. lateral flow assay) in POCT. In this review, we focus on the importance of using paper as a platform for sample pretreatment. We firstly discuss the beneficial use of paper for sample pretreatment, including sample collection and storage, separation, extraction, and concentration. We highlight the working principle and fabrication of each sample pretreatment device, the existing challenges and the future perspectives for developing paper-based sample pretreatment technique.

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