Animals have evolved defense systems for surviving in a chemically diverse environment. Such systems should demonstrate plasticity, such as adaptive immunity, enabling a response to even unknown chemicals. The antioxidant transcription factor Nrf2 is activated in response to various electrophiles and induces cytoprotective enzymes that detoxify them. We report here the discovery of a multiple sensing mechanism for Nrf2 activation using zebrafish and 11 Nrf2-activating compounds. First, we showed that six of the compounds tested specifically target Cys-151 in Keap1, the ubiquitin ligase for Nrf2, while two compounds target Cys-273. Second, in addition to Nrf2 and Keap1, a third factor was deemed necessary for responding to three of the compounds. Finally, we isolated a zebrafish mutant defective in its response to seven compounds but not in response to the remaining four. These results led us to categorize Nrf2 activators into six classes and hypothesize that multiple sensing allows enhanced plasticity in the system.Nrf2 is a transcription factor that transactivates cytoprotective genes through a common DNA regulatory element, called the antioxidant response element or electrophile response element (18, 24). Nrf2 target genes are multifarious and encode phase 2 detoxifying enzymes, antioxidant proteins, enzymes for glutathione biosynthesis, ABC transporters, scavenger receptors, transcription factors, proteases, chaperone proteins, and so forth (23). Under basal conditions, Nrf2 is rapidly degraded by proteasomes, and little induction of target genes is observed. This degradation is controlled by Keap1, an Nrf2-specific adaptor protein for the Cul3 ubiquitin ligase complex (12,20). Nrf2-activating compounds block Keap1-dependent Nrf2 ubiquitination, leading to the stabilization and nuclear translocation of Nrf2 and subsequent induction of Nrf2 target genes.A number of Nrf2 activators have been found but, interestingly, no common structures were identified among them (23). Talalay and coworkers classified Nrf2-activating compounds into the following 10 distinct classes based on their chemical structures (7): diphenols, Michael reaction acceptors, isothiocyanates, thiocarbamates, trivalent arsenicals, 1,2-dithiole-3-thiones, hydroperoxides, vicinal dimercaptans, heavy metals, and polyenes. A current pursuit is unraveling how cells detect these chemical compounds and transduce their signals into the activation of Nrf2. Keap1 has many highly reactive cysteine residues that have the potential to sense electrophilic Nrf2 activators by forming covalent adducts with them. We and others have therefore proposed the model that Nrf2-activating compounds directly modify the sulfhydryl groups of Keap1 cysteines by oxidation, reduction, or alkylation, which alters the conformation of Keap1 and ceases the ubiquitination of Nrf2 (7,24). In fact, mass spectrometry (MS) studies revealed that some Nrf2-activating compounds can covalently react with cysteines in mouse or human Keap1. For example, dexamethasone 21-mesylate with ; iodo...

Gonadotropin-releasing hormone (GnRH) is important in reproduction, although the mechanism of central hypogonadism in humans remains unclear. Because the GnRH neuron originates from the olfactory placode and migrates to the hypothalamus during development, central hypogonadism in humans could be caused by failure in normal migration of GnRH neurons to the hypothalamus. We report that in transgenic mice expression of the simian virus 40 T antigen, driven by the promoter of human GnRH gene, resulted in central hypogonadism due to an arrest in neuronal migration during development and tumor formation along the migratory pathway. This system appears to be an important animal model of hypogonadotropic hypogonadism in humans. Additionally, olfactory bulb tumors from these animals were dispersed, and a GnRH-secreting neuronal cell line (GN cell line) was established.

SummaryThe molecular mechanisms of plant responses to iron (Fe) de®ciency remain largely unknown. To identify the cis-acting elements responsible for Fe-de®ciency-inducible expression in higher plants, the barley IDS2 (iron de®ciency speci®c clone no. 2) gene promoter was analyzed using a transgenic tobacco system. Deletion analysis revealed that the sequence between À272 and À91 from the translational start site (À272/À91) was both suf®cient and necessary for speci®c expression in tobacco roots. Further deletion and linker-scanning analysis of this region clearly identi®ed two cis-acting elements: iron-de®ciency-responsive element 1 (IDE1) at À153/À136 (ATCAAGCATGCTTCTTGC) and IDE2 at À262/À236 (TTGAACGG-CAAGTTTCACGCTGTCACT). The co-existence of IDE1 and IDE2 was essential for speci®c expression when the À46/ 8 region (relative to the transcriptional start site) of the CaMV 35S promoter was used as a minimal promoter. Expression occurred mainly in the root pericycle, endodermis, and cortex. When the À90/ 8 region of the CaMV 35S promoter was fused, the À272/À227 region, which consists of IDE2 and an additional 19 bp, could drive Fe-de®ciency-inducible expression without IDE1 throughout almost the entire root. The principal modules of IDE1 and IDE2 were homologous. Sequences homologous to IDE1 were also found in many other Fe-de®ciency-inducible promoters, including: nicotianamine aminotransferase (HvNAAT)-A, HvNAAT-B, nicotianamine synthase (HvNAS1), HvIDS3, OsNAS1, OsNAS2, OsIRT1, AtIRT1, and AtFRO2, suggesting the conservation of cis-acting elements in various genes and species. The identi®cation of novel cis-acting elements, IDE1 and IDE2, will provide powerful tools to clarify the molecular mechanisms regulating Fe homeostasis in higher plants.

The objective of this study was to quantify the mastication effort for cooked rice. We analyzed mastication patterns while normal subjects ate a spoonful of cooked rice that had been prepared by cooking with different amounts of water (1.5, 2.0, 3.0, and 4.0 times the water to rice weight). The rice samples were served with the same weight, same volume and same solid content, and electromyography (EMG) of the masticatory muscles was measured. The texture of the four cooked rice samples was instrumentally analyzed by the two-bite method. The number of chews, masticatory time, and jaw-closing muscle activities per chew evaluated by EMG were higher in the rice sample cooked with least water, which exhibited a high firmness value in the instrumental test. Rice cooked with 4.0 times the amount of water exhibited the longest jaw-opening duration, which was related to the adhesiveness value in the instrumental test. The ratio of jaw-opening muscle activity to the preceding jaw-closing muscle activity was lower for the rice containing least water, this corresponding to the area ratio (balance degree) in the instrumental test. Softer rice containing more water reduced the total mastication effort until swallowing because it required a shorter mastication time. It was not difficult for the softer rice with high density to be ingested in greater weight, decreasing the mastication effort for a certain amount.