DYF-5/MAK-dependent phosphorylation promotes ciliary tubulin unloading.

Cilia are microtubule-based organelles that power cell motility and regulate sensation and signaling, and abnormal ciliary structure and function cause various ciliopathies. Cilium formation and maintenance requires intraflagellar transport (IFT), during which the kinesin-2 family motor proteins ferry IFT particles carrying axonemal precursors such as tubulins into cilia. Tubulin dimers are loaded to IFT machinery through an interaction between tubulin and the IFT-74/81 module; however, little is known of how tubulins are unloaded when arriving at the ciliary tip. Here, we show that the ciliary kinase DYF-5/MAK phosphorylates multiple sites within the tubulin-binding module of IFT-74, reducing the tubulin-binding affinity of IFT-74/81 approximately sixfold. Ablation or constitutive activation of IFT-74 phosphorylation abnormally elongates or shortens sensory cilia in Caenorhabditis elegans neurons. We propose that DYF-5/MAK-dependent phosphorylation plays a fundamental role in ciliogenesis by regulating tubulin unloading.

Reconstitution reveals motor activation for intraflagellar transport.

The human body represents a notable example of ciliary diversification. Extending from the surface of most cells, cilia accomplish a diverse set of tasks. Predictably, mutations in ciliary genes cause a wide range of human diseases such as male infertility and blindness. In Caenorhabditis elegans sensory cilia, this functional diversity appears to be traceable to the differential regulation of the kinesin-2-powered intraflagellar-transport (IFT) machinery. Here we reconstituted the first, to our knowledge, functional multi-component IFT complex that is deployed in the sensory cilia of C. elegans. Our bottom-up approach revealed the molecular basis of specific motor recruitment to the IFT trains. We identified the key component that incorporates homodimeric kinesin-2 into its physiologically relevant context, which in turn allosterically activates the motor for efficient transport. These results will enable the molecular delineation of IFT regulation, which has eluded understanding since its discovery more than two decades ago.

Hydrochemical Analysis and Fuzzy Logic Method for Evaluation of Groundwater Quality in the North Chengdu Plain, China.

Groundwater is a major water resource in the North Chengdu Plain, China. The research objective is to determine the quality and suitability of groundwater for drinking purposes within the vicinity of a shallow, unconsolidated aquifer of Quaternary age. In this study, a detailed investigation was conducted to define the hydrochemical characteristics that control the quality of groundwater, based on traditional methods. Considering the uncertainties linked with water resources and the environmental complications, the fuzzy logic method was used in the determination of groundwater quality for more precise findings that support decision-making. To achieve such an objective, sixteen water quality guidelines were used to determine groundwater quality status in six selected wells. The results showed that the groundwater is neutral, very hard, and fresh in nature. Dominating cations and anions are in the order of Ca 2 + > Na + > Mg 2 + > K + and HCO 3 - > SO 4 2 - > Cl - . The Piper trilinear diagram demonstrates that the hydrochemical facies of groundwater are mostly of Ca-HCO 3 type. Statistical analysis denotes a positive correlation between most of the chemical parameters. The study took the results of the fuzzy logic evaluation method into consideration, to classify the samples into five groups according to the Chinese groundwater quality standard (GB/T 14848-93) for their suitability for domestic use. The results demonstrated that the quality of the groundwater samples is within grade II and III, and is suitable for drinking purposes. The comprehensive evaluation of groundwater quality is critical to aid sensitive policy decisions, and the proposed approach can guarantee reliable findings to that effect. The results of this study would also be helpful to future researches related to groundwater quality assessment.

A GIS-based DRASTIC model for assessing aquifer vulnerability in Kakamigahara Heights, Gifu Prefecture, central Japan.

Vulnerability assessment to delineate areas that are more susceptible to contamination from anthropogenic sources has become an important element for sensible resource management and land use planning. This contribution aims at estimating aquifer vulnerability by applying the DRASTIC model as well as utilizing sensitivity analyses to evaluate the relative importance of the model parameters for aquifer vulnerability in Kakamigahara Heights, Gifu Prefecture central Japan. An additional objective is to demonstrate the combined use of the DRASTIC and geographical information system (GIS) as an effective method for groundwater pollution risk assessment. The DRASTIC model uses seven environmental parameters (Depth to water, net Recharge, Aquifer media, Soil media, Topography, Impact of vadose zone, and hydraulic Conductivity) to characterize the hydrogeological setting and evaluate aquifer vulnerability. The western part of the Kakamigahara aquifer was dominated by "High" vulnerability classes while the eastern part was characterized by "Moderate" vulnerability classes. The elevated north-eastern part of the study area displayed "Low" aquifer vulnerability. The integrated vulnerability map shows the high risk imposed on the eastern part of the Kakamigahara aquifer due to the high pollution potential of intensive vegetable cultivation. The more vulnerable western part of the aquifer is, however, under a lower contamination risk. In Kakamigahara Heights, land use seems to be a better predictor of groundwater contamination by nitrate. Net recharge parameter inflicted the largest impact on the intrinsic vulnerability of the aquifer followed by soil media, topography, vadose zone media, and hydraulic conductivity. Sensitivity analyses indicated that the removal of net recharge, soil media and topography causes large variation in vulnerability index. Moreover, net recharge and hydraulic conductivity were found to be more effective in assessing aquifer vulnerability than assumed by the DRASTIC model. The GIS technique has provided efficient environment for analyses and high capabilities of handling large spatial data.

Assessment of groundwater contamination by nitrate leaching from intensive vegetable cultivation using geographical information system.

This study employed the Geographical Information System (GIS) technology to investigate nitrate contamination of groundwater by agrochemical fertilizers in the Kakamigahara Heights, Gifu Prefecture, central Japan. Thematic information and chemical data of groundwater from the Heights were analyzed in a GIS environment to study the extent and variation of nitrate contamination and to establish spatial relationships with responsible land use types. The high and correlated concentrations of Ca(2+), Mg(2+), SO(4)(2-), and NO(3)(-) reflected the polluted nature of the unconfined highly permeable Kakamigahara aquifer. Ninety percent of the water samples showed nitrate concentrations above the human affected value (3 mg/l NO(3)(-)), while more than 30% have exceeded the maximum acceptable level (44 mg/l NO(3)(-)) according to Japan regulations. The spatial analyses indicated that groundwater contamination by nitrate is closely associated with one specific land use class, the "vegetable fields". The nitrate concentration of groundwater under vegetable fields was significantly higher than that under urban land or paddy fields. Most of the unacceptable nitrate levels were encountered in boreholes assigned to "vegetable fields" but a few were also found in boreholes allotted to "urban" class. Therefore, the vegetable fields were considered the principal source of nitrate contamination of groundwater in the Kakamigahara. However, contamination from urban sources is also possible.

Natural denitrification in the Kakamigahara groundwater basin, Gifu prefecture, central Japan.

Although nitrate is recognized as the most common groundwater contaminant due to growing anthropogenic sources, such as agriculture in particular, its adverse effects on human and animal health are debatable. The current issue, however, is to control and reduce nitrate contamination with regards to the long residence time of groundwater within aquifers. Denitrification has recently been recognized for its ability to reduce high nitrate concentrations in groundwater. The Kakamigahara groundwater basin, Gifu prefecture, Japan, witnessed rising levels of nitrate (>12 mg/l NO(3)-N) originating from agricultural sources. Chemical analyses for the determination of major constituents of groundwater and delta(15)N of residual nitrate were performed on representative groundwater samples in order to fulfill two main objectives. One is to investigate the current situation of nitrate groundwater pollution. The second objective is to determine whether the denitrification is a potential natural mechanism, which eliminates nitrate pollution in the Kakamigahara aquifer. Agricultural nitrate contamination of groundwater was obvious from characteristically high concentrations of Ca(2+), Mg(2+), NO(3)(-) and SO(4)(2-). High nitrate concentrations were found on the eastern side of the basin in association with vegetable cultivation fields, and decreased gradually towards the west of the basin along the direction of groundwater flow. The decrease of nitrate concentration was conveniently coupled with increase of HCO(3)(-) (the heterotrophic denitrification product), pH and delta(15)N of residual nitrate (due to isotopic fractionation) from east to west. Therefore, denitrification in situ is continuously removing nitrate from the Kakamigahara groundwater system.

Histomolecular phenotypes and outcome in adenocarcinoma of the ampulla of vater.

PURPOSE: Individuals with adenocarcinoma of the ampulla of Vater demonstrate a broad range of outcomes, presumably because these cancers may arise from any one of the three epithelia that converge at that location. This variability poses challenges for clinical decision making and the development of novel therapeutic strategies. PATIENTS AND METHODS: We assessed the potential clinical utility of histomolecular phenotypes defined using a combination of histopathology and protein expression (CDX2 and MUC1) in 208 patients from three independent cohorts who underwent surgical resection for adenocarcinoma of the ampulla of Vater. RESULTS: Histologic subtype and CDX2 and MUC1 expression were significant prognostic variables. Patients with a histomolecular pancreaticobiliary phenotype (CDX2 negative, MUC1 positive) segregated into a poor prognostic group in the training (hazard ratio [HR], 3.34; 95% CI, 1.69 to 6.62; P < .001) and both validation cohorts (HR, 5.65; 95% CI, 2.77 to 11.5; P < .001 and HR, 2.78; 95% CI, 1.25 to 7.17; P = .0119) compared with histomolecular nonpancreaticobiliary carcinomas. Further stratification by lymph node (LN) status defined three clinically relevant subgroups: one, patients with histomolecular nonpancreaticobiliary (intestinal) carcinoma without LN metastases who had an excellent prognosis; two, those with histomolecular pancreaticobiliary carcinoma with LN metastases who had a poor outcome; and three, the remainder of patients (nonpancreaticobiliary, LN positive or pancreaticobiliary, LN negative) who had an intermediate outcome. CONCLUSION: Histopathologic and molecular criteria combine to define clinically relevant histomolecular phenotypes of adenocarcinoma of the ampulla of Vater and potentially represent distinct diseases with significant implications for current therapeutic strategies, the ability to interpret past clinical trials, and future trial design.