EML4-ALK fusion genes were observed predominantly in adenocarcinomas, in female or nonsmoking populations. Additionally, the EML4-ALK fusions were mutually exclusive with mutations in the EGFR, KRAS, and ERBB2 genes.

A new automated shipboard analytical method for determining iron(III) in seawater has been developed. The method is based on a combination of selective column extraction using chelating resin and improved chemiluminescence (CL) detection in a closed flowthrough system. In this method, Fe(III) in an acidified sample solution is selectively collected on 8-quinolinol immobilized chelating resin and then eluted with dilute hydrochloric acid. The resulting eluent is mixed with luminol solution, aqueous ammonia, and hydrogen peroxide solution successively, and then the mixture is introduced into the CL cell. The iron concentration is obtained from the CL intensity. The detection limit of iron(III) is 0.05 nmol L1 234when using an 18-mL seawater sample. The method was applied to ordinary oceanic waters and hydrothermal waters collected in the North and South Pacific Oceans. Worldwide marine chemists and marine biologists have focused on the behavior of iron in seawater, since Martin et al.1"5 pointed out that the phytoplankton growth in oceanic water was limited by the deficiency of iron derived from the atmosphere rather than the lack of nutrients in some oceanic regions, such as the equatorial Pacific, Gulf of Alaska, and Antarctic Ocean. This attractive hypothesis created a heated argument in various journals6 78"9 and spurred the geochemical study of iron. For example, Zhuang et al.10 reported recently that more than half of the iron in aeolian mineral dust existed in the form of Fe (II), resulting in the enhancement of solubility (1) Martin,

The small GTPases RalA and RalB are multifunctional proteins regulating a variety of cellular processes. Like other GTPases, the activity of Ral is regulated by the opposing effects of guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs). Although several RalGEFs have been identified and characterized, the molecular identity of RalGAP remains unknown. Here, we report the first molecular identification of RalGAPs, which we have named RalGAP1 and RalGAP2. They are large heterodimeric complexes, each consisting of a catalytic alpha1 or alpha2 subunit and a common beta subunit. These RalGAP complexes share structural and catalytic similarities with the tuberous sclerosis tumor suppressor complex, which acts as a GAP for Rheb. In vitro GTPase assays revealed that recombinant RalGAP1 accelerates the GTP hydrolysis rate of RalA by 280,000-fold. Heterodimerization was required for this GAP activity. In PC12 cells, knockdown of the beta subunit led to sustained Ral activation upon epidermal growth factor stimulation, indicating that the RalGAPs identified here are critical for efficient termination of Ral activation induced by extracellular stimuli. Our identification of RalGAPs will enable further understanding of Ral signaling in many biological and pathological processes.

No abstract

Keap1 gene mutations are likely to be associated with a worse prognosis and lower postoperative disease-free survival rates in pathological Stage I-II NSCLC.

BackgroundDownstream activation through receptor tyrosine kinases (RTKs) plays important roles in carcinogenesis. In this study, we assessed the clinical involvement of Axl, an RTK, and its ligand, Gas6, in surgically treated lung adenocarcinoma.MethodsAxl and Gas6 mRNA and protein expression levels were quantified using quantitative real-time polymerase chain reaction and immunohistochemistry, respectively, in completely resected lung adenocarcinoma tissues (n = 88) and were evaluated for correlation with clinicopathologic features and patient survival.ResultsHigher expressions of Axl mRNA/protein and Gas6 protein were significantly related to worse clinicopathological features and prognosis (5-year overall survival rates: Axl mRNA low: 72.3 %, high: 49.7 %, P = 0.047; Axl protein low: 77.5 %, high: 38.6 %, P < 0.001; and Gas6 protein low: 70.5 %, high: 48 %, P = 0.042). On the contrary, higher Gas6 mRNA expression was related to better clinicopathological features and prognosis (5-year overall survival rates: Gas6 mRNA low: 59.2 %, high: 81.8 %, P = 0.054). Multivariate analysis suggests that high Axl mRNA expression may be an independent factor for poor patient prognosis (P = 0.04).ConclusionsIn lung adenocarcinoma, Axl and Gas6 expression levels were associated with tumor advancement and patient survival, thus rendering them as reliable biomarkers and potential targets for treatment of lung adenocarcinoma.

The small GTPase Ral is known to be highly activated in several human cancers, such as bladder, colon and pancreas cancers. It is reported that activated Ral is involved in cell proliferation, migration and metastasis of bladder cancer. This protein is activated by Ral guanine nucleotide exchange factors (RalGEFs) and inactivated by Ral GTPase-activating proteins (RalGAPs), the latter of which consist of heterodimers containing a catalytic α1 or α2 subunit and a common β subunit. In Ras-driven cancers, such as pancreas and colon cancers, constitutively active Ras mutant activates Ral through interaction with RalGEFs, which contain the Ras association domain. However, little is known with regard to the mechanism that governs aberrant activation of Ral in bladder cancer, in which Ras mutations are relatively infrequent. Here, we show that Ral was highly activated in invasive bladder cancer cells due to reduced expression of RalGAPα2, the dominant catalytic subunit in bladder, rather than increased expression of RalGEFs. Exogenous expression of wild-type RalGAPα2 in KU7 bladder cancer cells with invasive phenotype, but not mutant RalGAPα2-N1742K lacking RalGAP activity, resulted in attenuated cell migration in vitro and lung metastasis in vivo. Furthermore, genetic ablation of Ralgapa2 promoted tumor invasion in a chemically-induced murine bladder cancer model. Importantly, immunohistochemical analysis of human bladder cancer specimens revealed that lower expression of RalGAPα2 was associated with advanced clinical stage and poor survival of patients. Collectively, these results are highly indicative that attenuated expression of RalGAPα2 leads to disease progression of bladder cancer through enhancement of Ral activity.

Diel changes in bacterial ingestion by a mixotrophic flagellate, Cryptomonas sp., and heterotrophic nanoflagellates (HNF) were examined in situ at 4-h intervals for 2 d in the epilimnion and metalimnion of Lake Biwa using bacteria-sized fluorescent microspheres as a tracer food. Clearance rates of HNF for the microspheres ranged between 1.3 and 4.5 nl cell Ϫ1 h Ϫ1 , but the average rate did not differ between day and night. In contrast, clear diel changes were observed in the clearance rate of Cryptomonas sp. in the epilimnion from Ͻ0.5 nl cell Ϫ1 h Ϫ1 at midnight to Ͼ3 nl cell Ϫ1 h Ϫ1 at noon. In the metalimnion where light intensity was lower, however, the clearance rate of Cryptomonas sp. was always Ͻ0.5 nl cell Ϫ1 h Ϫ1 through the study period. Thus, bacterial ingestion of Cryptomonas sp. is not to acquire supplementary energy or carbon at low phototrophic activities. During the study period, both inorganic phosphorus and nitrogen concentrations were less than or close to the detection limits (10 nM P and 1 M N) in the epilimnion, but much higher in the metalimnion. The results strongly support the idea that Cryptomonas sp. utilizes N and P from bacteria as substitutable nutrients when photosynthesis takes place under conditions of nutrient depletion. To assess the grazing effect of mixotrophic algae on bacterial populations, it is essential to consider diel changes in their phagotrophic mode of nutrition that are induced by light regime and nutrient concentrations in ambient water.