In mammals and birds, thermoregulation to conserve body temperature is vital to life. Multiple mechanisms of thermogeneration have been proposed, localized in different subcellular organelles. However, visualizing thermogenesis directly in intact organelles has been challenging. Here we have developed genetically encoded, GFP-based thermosensors (tsGFPs) that enable visualization of thermogenesis in discrete organelles in living cells. In tsGFPs, a tandem formation of coiled-coil structures of the Salmonella thermosensing protein TlpA transmits conformational changes to GFP to convert temperature changes into visible and quantifiable fluorescence changes. Specific targeting of tsGFPs enables visualization of thermogenesis in the mitochondria of brown adipocytes and the endoplasmic reticulum of myotubes. In HeLa cells, tsGFP targeted to mitochondria reveals heterogeneity in thermogenesis that correlates with the electrochemical gradient. Thus, tsGFPs are powerful tools to noninvasively assess thermogenesis in living cells.

Laminin-5, consisting of the ␣3, ␤3, and ␥2 chains, is localized in the skin basement membrane and supports the structural stability of the epidermo-dermal linkage and regulates various cellular functions. The ␣ chains of laminins have been shown to have various biological activities. In this study, we identified a sequence of the ␣3 chain C-terminal globular domain (LG1-LG5 modules) required for both heparin binding and cell adhesion using recombinant proteins and synthetic peptides. We found that the LG3 and LG4 modules have activity for heparin binding and that LG4 has activity for cell adhesion. Studies with synthetic peptides delineated the A3G75aR sequence (NSFMALYLSKGR, residues 1412-1423) within LG4 as a major site for both heparin and cell binding. Substitution mutations in LG4 and A3G75aR identified the Lys and Arg of the A3G75aR sequence as critical for these activities. Cell adhesion to LG4 and A3G75aR was inhibited by heparitinase I treatment of cells, suggesting that cell binding to the A3G75aR site was mediated by cell surface heparan sulfate proteoglycans. We showed by affinity chromatography that syndecan-2 from fibroblasts bound to LG4. Solid-phase assays confirmed that syndecan-2 interacted with the A3G75aR peptide sequence. Stably transfected 293T cells with expression vectors for syndecan-2 and -4, but not glypican-1, specifically adhered to LG4 and A3G75aR. These results indicate that the A3G75aR sequence within the laminin ␣3 LG4 module is responsible for cell adhesion and suggest that syndecan-2 and -4 mediate this activity.Laminins are extracellular proteins primarily present at the basement membrane, where they provide structural stability and exert many biological functions, including cell adhesion, migration, proliferation, and differentiation, and they are also involved in angiogenesis and tumor invasion (for reviews, see Refs. 1 and 2). Laminins are a large family of glycoproteins, consisting of at least 12 isoforms (laminin-1 to -12) (3-5). Each laminin is composed of three different chains, ␣, ␤, and ␥. There are 11 laminin chains, five ␣ chains (␣1-␣5), three ␤ chains (␤1-␤3), and three ␥ chains (␥1-␥3), whose expression is regulated temporarily and specially during development and tissue repair. Three chains are assembled into a cross-shaped heterotrimer (␣␤␥) by forming a triple-stranded coiled-coil structure through the ␣-helical domain of each chain (6, 7).Laminin-5 (␣3␤3␥2) is a component of anchoring fibrils and forms a complex with the hemidesmosome apparatus and stabilizes the basement membrane structure by forming supramolecular complexes with laminin-6 and -7, collagen VII (8, 9), and fibulin-2 (10). Mutations in laminin-5 (11, 12) cause congenital skin blister diseases junctional epidermolysis bullosa. Disruption of the laminin ␣3 gene in mice resulted in abnormal hemidesmosomes and blister formation in the skin (13). Laminin-5 is also shown to be an adhesive substrate for keratinocytes (14). In these processes, the ␣ 3 ␤ 1 integrin was identified as a cellular recepto...

The KCNE1-D85N polymorphism was significantly more frequent in our LQTS probands. The functional variant is a disease-causing gene variant of LQTS phenotype that functions by interacting with KCNH2 and KCNQ1. Since its allele frequency was approximately 1% among control individuals, KCNE1-D85N may be a clinically important genetic variant.

Direct observations of the upper ocean velocity in the eastern equatorial Indian Ocean by an acoustic Doppler current profiler, from November 2000 to October 2001 on the equator at 90°E, demonstrate that the dominant periods of variability in the upper layer zonal and meridional currents are in intraseasonal frequency bands with periods of 30 to 50 days and 10 to 20 days, respectively. The strong intraseasonal variability in the zonal current obscures the semiannual Wyrtki jets, which can be seen clearly in the monthly averaged field. In addition, a zone of strong vertical shear of the zonal current and a distinct Equatorial Undercurrent with semiannual period are observed. The results provide us with a new perspective on importance of the energetic intraseasonal variability in the eastern equatorial Indian Ocean, which indicates strong correlation with the wind variability near the mooring location.

Subclinical I(Ks) dysfunction could be a risk of drug-induced TdP. Verapamil is effective in decreasing the QT interval and TDR and in suppressing EADs, thus preventing TdP in the model of acquired LQTS.