Spinesin/TMPRSS5, a novel transmembrane serine protease, cloned from human spinal cord.

A cDNA encoding a novel serine protease, which we designated spinesin, has been cloned from human spinal cord. The longest open reading frame was 457 amino acids. A homology search revealed that the human spinesin gene was located at chromosome 11q23 and contained 13 exons, the gene structure being similar to that of TMPRSS3 whose gene is also located on 11q23. Spinesin has a simple type II transmembrane structure, consisting of, from the N terminus, a short cytoplasmic domain, a transmembrane domain, a stem region containing a scavenger receptor-like domain, and a serine protease domain. Unlike TMPRSS3, it carries no low density lipoprotein receptor domain in the stem region. The extracellular region carries five N-glycosylation sites. The sequence of the protease domain carried the essential triad His, Asp, and Ser and showed some similarity to that of TMPRSS2, hepsin, HAT, MT-SP1, TMPRSS3, and corin, sharing 45.5, 41.9, 41.3, 40.3, 39.1, and 38.5% identity, respectively. The putative mature protease domain preceded by H(6)DDDDK was produced in Escherichia coli, purified, and successfully activated by immobilized enterokinase. Its optimal pH was about 10. It cleaved synthetic substrates for trypsin, which is inhibited by p-amidinophenylmethanesulfonyl fluoride hydrochloride but not by antipain or leupeptin. Northern blot analysis against mRNA from human tissues including liver, lung, placenta, and heart demonstrated a specific expression of spinesin mRNA in the brain. Immunohistochemically, spinesin was predominantly expressed in neurons, in their axons, and at the synapses of motoneurons in the spinal cord. In addition, some oligodendrocytes were clearly stained. These results indicate that spinesin is transported to the synapses through the axons after its synthesis in the cytoplasm and may play important roles at the synapses. Further analyses are required to clarify its roles at the synapses and in oligodendrocytes.     

The effects of endangered freshwater pearl mussels on channel morphology and flow in a low-gradient sandy river

Hydrobiologia - Conservation of ecosystem engineers, which modulates the surrounding habitat by causing physical state changes in biotic or abiotic materials, is important for maintaining the...     

Distribution dynamics of perfluorocarbon delivery to the lungs: an intact rabbit model.

Motivated by the goal of understanding how to most homogeneously fill the lungs with perfluorocarbon for liquid ventilation, we investigate the transport of liquid instilled into the lungs using an intact rabbit model. Perfluorocarbon is instilled into the trachea of the ventilated animal. Radiographic images of the perfluorocarbon distribution are obtained at a rate of 30 frames/s during the filling process. Image analysis is used to quantify the liquid distribution (center of mass, spatial standard deviation, skewness, kurtosis, and indicators of homogeneity) as time progresses. We compare the distribution dynamics in supine animals to those in upright animals for three constant infusion rates of perfluorocarbon: 15, 40, and 60 ml/min. It is found that formation of liquid plugs in large airways, which is affected by posture and infusion rate, can result in a more homogeneous liquid distribution than gravity drainage alone. The supine posture resulted in more homogeneous filling of the lungs than did upright posture, in which the lungs tend to fill in the inferior regions first. Faster instillation of perfluorocarbon results in liquid plugs forming in large airways and, consequently, more uniform distribution of perfluorocarbon than slower instillation rates in the upright animals.     

Comparative study of Agkistrodon acutus venoms from China and Taiwan

Three hemorrhagic toxins (Ac1-, Ac2- and Ac3-proteinases) were isolated from the lyophilized venom of Agkistrodon acutus from China using gel filtration on a Sephadex G-75 column, followed by chromatography on diethylaminoethyl (DEAE)-Sephadex A-50, DEAE-Sephacel and DEAE-cellulose. Homogeneity was established by the formation of a single band in acrylamide gel electrophoresis, isoelectric focusing and sodium dodecyl sulfate (SDS) acrylamide gel electrophoresis. Three hemorrhagic toxins possessed both lethal and proteolytic activities. These activities were inhibited by ethylenediamine tetraacetic acid (EDTA), ethyleneglycol (beta-aminoethyl)N,N,N',N'-tetraacetic acid (EGTA), o-phenanthroline or cysteine, but not by soybean trypsin inhibitor (SBTI), p-chloromercuribenzoate (PCMB) or diisopropyl fluorophosphate (DFP). Molecular weight of hemorrhagic toxins (Ac1-, Ac2- and Ac3-proteinases) were determined to be 24,500, 25,000 and 57,000, respectively. It was found that hemorrhagic toxins have considerable similarity to hemorrhagic toxins isolated by Nikai et al. (1977) and Sugihara et al. (1978, 1979), but Ac2- and Ac3-proteinases from China and Taiwan are electrophoretically unrelated.     

Carbon tetrachloride suppresses ER-Golgi transport by inhibiting COPII vesicle formation on the ER membrane in the RLC-16 hepatocyte cell line

Carbon tetrachloride (CCl₄) causes hepatotoxicity in mammals, with its hepatocytic metabolism producing radicals that attack the intracellular membrane system and destabilize intracellular vesicle transport. Inhibition of intracellular transport causes lipid droplet retention and abnormal protein distribution. The intracellular transport of synthesized lipids and proteins from the endoplasmic reticulum (ER) to the Golgi apparatus is performed by coat complex II (COPII) vesicle transport, but how CCl₄ inhibits COPII vesicle transport has not been elucidated. COPII vesicle formation on the ER membrane is initiated by the recruitment of Sar1 protein from the cytoplasm to the ER membrane, followed by that of the COPII coat constituent proteins (Sec23, Sec24, Sec13, and Sec31). In this study, we evaluated the effect of CCl₄ on COPII vesicle formation using the RLC-16 rat hepatocyte cell line. Our results showed that CCl₄ suppressed ER-Golgi transport in RLC-16 cells. Using a reconstituted system of rat liver tissue-derived cytoplasm and RLC-16 cell-derived ER membranes, CCl₄ treatment inhibited the recruitment of Sar1 and Sec13 from the cytosolic fraction to ER membranes. CCl₄-induced changes in the ER membrane accordingly inhibited the accumulation of COPII vesicle-coated constituent proteins on the ER membrane, as well as the formation of COPII vesicles, which suppressed lipid and protein transport between the ER and Golgi apparatus. Our data suggest that CCl₄ inhibits ER-Golgi intracellular transport by inhibiting COPII vesicle formation on the ER membrane in hepatocytes.     

Angular optimization for cancer identification with circularly polarized light

Depolarization of circularly polarized light scattered from biological tissues depends on structural changes in cell nuclei, which can provide valuable information for differentiating cancer tissues concealed in healthy tissues. In this study, we experimentally verified the possibility of cancer identification using scattering of circularly polarized light. We investigated the polarization of light scattered from a sliced biological tissue with various optical configurations. A significant difference between circular polarizations of light scattered from cancerous and healthy tissues is observed, which is sufficient to distinguish a cancerous region. The line-scanning experiments along a region incorporating healthy and cancerous parts indicate step-like behaviors in the degree of circular polarization corresponding to the state of tissues, whether cancerous or normal. An oblique and perpendicular incidence induces different resolutions for identifying cancerous tissues, which indicates that the optical arrangement can be selected according to the priority of resolution.     

Metabolism of three stereoisomers of astaxanthin in the fish, rainbow trout and tilapia

In rainbow trout (Salmo gairdneri), the dietary astaxanthin diesters were mostly absorbed and accumulated in their integuments keeping their configurations, and partially metabolized to (3R, 3'R)-zeaxanthin (major) and/or (3R, 3'S)-zeaxanthin (medium) and/or (3S,3'S)-zeaxanthin (minor). In tilapia (Tilapia nilotica), the three stereoisomers of astaxanthin diesters were promptly metabolized to only (3S,3'S)-astaxanthin, and subsequently to (3R,3'R)-zeaxanthin and/or (3R,3'S)-zeaxanthin and/or (3S,3'S)-zeaxanthin at an invariable ratio, 4:1:0.3. The above facts indicate that the conversion from 3S- to 3R-configuration was carried out in vivo, and vice versa, and that astaxanthins were reductively metabolized to zeaxanthins in both the fish.     

Nicorandil reduces the basal level of cytosolic free calcium in single guinea pig ventricular myocytes.

Using fluorescent Ca2+ indicator fura-2 and whole-cell patch-clamp techniques, we examined the effect of 2-nicotinamidoethyl nitrate (nicorandil) on the intracellular free Ca2+ concentration ([Ca2+]i) and electrical properties in single guinea pig ventricular myocytes. Nicorandil (10 nM approximately 1 mM) reduced the resting level [Ca2+]i monitored as fura-2 fluorescence ratio in a concentration-dependent manner. Dibutyryl guanosine 3':5'-cyclic monophosphate (cyclic GMP), a membrane permeable cyclic GMP analogue, mimicked the nicorandil action. Neither application of caffeine (10 mM) nor deprivation of extracellular Na+ ions could prevent the nicorandil action on [Ca2+]i. In contrast, the nicorandil effect was virtually blocked by sodium orthovanadate (40 microM), a Ca2+ pumping ATPase inhibitor. During electrophysiological experiments, nicorandil shortened action potential durations (205 +/- 80 ms to 153 +/- 76 ms) by increasing a glibenclamide-sensitive outward K+ conductance. However, the drug produced little hyperpolarization (approximately 2 mV) because the resting potential of ventricular myocytes was close to the K+ equilibrium potential. The involvement of voltage-dependent Ca-channel current and Na-Ca exchanger was considered to be minimal under physiological conditions. It is thus concluded that nicorandil decreases basal [Ca2+]i via cyclic GMP-mediated activation of the plasma membrane Ca2+ pump in guinea pig ventricular myocytes.     

Diel patterns in singing activity of humpback whales in a winter breeding area in Okinawan (Ryukyuan) waters

Male humpback whales produce complex sounds known as songs during their breeding season. Previous studies have shown diel patterns of song in their breeding areas, but there had been no similar studies in the breeding area around Okinawa, Japan. To study diel patterns of song and the behavior of humpback whales in Okinawa, we conducted 24 hr recording with a fixed hydrophone in 2007, and vessel-based sighting surveys during 2014–2017. Song was monitored for 15 days, with peaks at sunrise and around 2200. Singing activity declined significantly between sunrise and sunset, then increased until 2200. Activity levels at night were higher and more stable than during the day. During 278 days of sighting surveys, 2,551 whales in 1,382 groups were observed. 79 individuals were confirmed as singers, all of which were lone whales. In six cases, singing individuals stopped singing before joining a group or began singing after leaving a group. Previous studies have shown that group size of humpback whales increases through the day. Considering the results from our study and the former studies, the decrease in singing activity as the day progresses may be a result of aggregation increasing, thus reducing the number of lone singers during the day.