Chlorine sensitivity of feline calicivirus, a norovirus surrogate

The sensitivity to free chlorine of feline calicivirus (FCV), a norovirus surrogate, was examined relative to chlorine demand. When a crude suspension of FCV was treated with a sodium hypochlorite solution containing 10 microg/ml free chlorine, the extent of the decrease of viral infectivity clearly depended on the volume of the reaction mixture. The apparent sensitivity of FCV to free chlorine increased with the reduction of host cell debris, indicating that chlorine demand must be minimized to know the true sensitivity of the virus. We therefore partially purified the viruses from the host cell components and found that the infectivity of FCV was reduced by more than log 4.6 by 5 min of treatment with 300 ng/ml free chlorine.     

Anticipatory Cortical Activation Precedes Auditory Events in Sleeping Infants

Background

Behavioral studies have shown that infants can form associations between environmental events and produce anticipatory actions for the predictable event, but the neural mechanisms for the learning and anticipation of events in infants are not known. Recent neuroimaging studies revealed that the association cortices of infants show activation related to auditory-stimulus discrimination and novelty detection during sleep. In the present study, we expected that when an auditory cue (beeps) predicted an auditory event (a female voice), specific regions of the infant cortex would show anticipatory activation before the event onset even while sleeping.

Methodology/Principal Findings

We examined the cortical activation of 3-month-old infants during delays between the cue and the event by using multi-channel near-infrared spectroscopy. To investigate spatiotemporal changes in cortical activation over the experimental session, we divided the session into two phases (early and late phase) and analyzed each phase separately. In the early phase, the frontal regions showed activation in response to the cue that was followed by the event compared with another cue that was not followed by any event. In the late phase, the temporoparietal region, in addition to the frontal region, showed prominent activation in response to the cue followed by the event. In contrast, when the cue was followed by an event and no-event in equal proportions, cortical activation in response to the cue was not observed in any phase.

Conclusions

Sleeping 3-month-old infants showed anticipatory cortical activation in the temporoparietal and frontal regions only in response to the cue predicting the event, suggesting that infants can implicitly form associations between temporally separated events and generate the anticipatory activation before the predictable event. Furthermore, the different time evolution of activation in the temporoparietal and frontal regions suggests that these regions may be involved in different aspects of learning and predicting future events.     

Stabilizing effect of cholesterol on phosphatidylcholine vesicles observed by ultrasonic velocity measurement.

The temperature dependence of the ultrasonic velocity was measured in sonicated vesicles of dipalmitoylphosphatidylcholine by varying the content of cholesterol. When cholesterol is incorporated, an anomalous dip of the ultrasonic velocity gradually smeared out. At the same time, the ultrasonic velocity of the membrane increased remarkably above 30 degrees C due to the increase of the bulk modulus by about 15%. On the other hand, the ultrasonic velocity and the bulk modulus decreased below 30 degrees C. Comparing the cholesterol-incorporated membrane with vesicles of bovine brain sphingomyelin and human erythrocyte membrane, we discuss the role of cholesterol in biological membranes in terms of the stability of the membrane as a barrier.     

Inhibition of IGF-1-Mediated Cellular Migration and Invasion by Migracin A in Ovarian Clear Cell Carcinoma Cells

Previously we isolated migracin A from a Streptomyces culture filtrate as an inhibitor of cancer cell migration. In the present research, we found that migracin A inhibited migration and invasion of ovarian clear cell carcinoma ES-2 cells. In the course of our mechanistic study, migracin A was shown to enhance vasohibin-1 expression in an angiogenesis array. We also confirmed that it increased the mRNA expression of this protein. Moreover, overexpression of vasohibin-1 lowered the migration but not the invasion of ES-2 cells. Then, we looked for another target protein employing a motility array, and found that migracin A lowered the IGF-1 expression. Knockdown of IGF-1 by siRNA decreased the migration and invasion of ES-2 cells. Migracin A also decreased Akt phosphorylation involved in the downstream signaling. Crosstalk analysis indicated that overexpression of vasohibin-1 decreased the IGF-1 expression. On the other hand, it showed no direct anticancer activity in terms of the ES-2 growth in agar. Migracin A inhibited the migration and IGF-1 expression in not only ES-2 but also another ovarian clear cell carcinoma JHOC-5 cells. In addition, it also inhibited capillary tube formation of human umbilical vein endothelial cells. Since its cytotoxicity is very low, migracin A may be a candidate for an anti-metastasis agent not exhibiting prominent toxicity.     

The Roles of ROS and Caspases in TRAIL-Induced Apoptosis and Necroptosis in Human Pancreatic Cancer Cells

Death signaling provided by tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) can induce death in cancer cells with little cytotoxicity to normal cells; this cell death has been thought to involve caspase-dependent apoptosis. Reactive oxygen species (ROS) are also mediators that induce cell death, but their roles in TRAIL-induced apoptosis have not been elucidated fully. In the current study, we investigated ROS and caspases in human pancreatic cancer cells undergoing two different types of TRAIL-induced cell death, apoptosis and necroptosis. TRAIL treatment increased ROS in two TRAIL-sensitive pancreatic cancer cell lines, MiaPaCa-2 and BxPC-3, but ROS were involved in TRAIL-induced apoptosis only in MiaPaCa-2 cells. Unexpectedly, inhibition of ROS by either N-acetyl-L-cysteine (NAC), a peroxide inhibitor, or Tempol, a superoxide inhibitor, increased the annexin V-/propidium iodide (PI)⁺ early necrotic population in TRAIL-treated cells. Additionally, both necrostatin-1, an inhibitor of receptor-interacting protein kinase 1 (RIP1), and siRNA-mediated knockdown of RIP3 decreased the annexin V^-/PI⁺ early necrotic population after TRAIL treatment. Furthermore, an increase in early apoptosis was induced in TRAIL-treated cancer cells under inhibition of either caspase-2 or -9. Caspase-2 worked upstream of caspase-9, and no crosstalk was observed between ROS and caspase-2/-9 in TRAIL-treated cells. Together, these results indicate that ROS contribute to TRAIL-induced apoptosis in MiaPaCa-2 cells, and that ROS play an inhibitory role in TRAIL-induced necroptosis of MiaPaCa-2 and BxPC-3 cells, with caspase-2 and -9 playing regulatory roles in this process.     

Characterization of thallus mechanical and physiological traits of tropical fucoids: A preliminary study

Thallus mechanical strength, as well as production of secondary metabolites as defensive compounds, of tropical macroalgae is often essential for protection from herbivory. In tropical macroalgae, thallus mechanical strength is negatively correlated with productivity — a trade‐off between productivity and thallus toughness. The tropical fucoids, Turbinaria ornata which has defensive morphological traits against herbivory and Sargassum ilicifolium which is expanding its distribution in Japanese waters, were examined to determine thallus traits related to mechanical strength and productivity and their pair‐wise relationships were also examined. These traits that are directly or indirectly related to the trade‐off between productivity and thallus toughness were compared to data for various other temperate macroalgae by regression analyses. We found two strong positive correlations between thallus mechanical strength and thallus mass or thallus thickness, confirming that higher levels of mechanical strength for tropical fucoids is associated with higher biomass or thallus thickness. Also, negative correlations between thallus toughness and productivity were found indicating structural and physiological trade‐offs. However, the tropical fucoids exhibited relatively high productivity regardless of their higher level of thallus toughness. These traits of the tropical fucoids slightly deviate from the typical conservative strategy with higher thallus toughness and lower productivity as a trade‐off between productivity and thallus toughness.     

Dual signal-responsive liposomes for temperature-controlled cytoplasmic delivery

For production of a new type of functional liposome whose destabilization can be triggered by a combination of a temperature signal and acidic pH signal, we prepared liposomes modified with hyperbranched poly(glycidol) derivatives having N-isopropylamide and carboxyl groups. HeLa cells incubated with the dual signal-responsive liposomes encapsulating a water-soluble fluorescent dye pyranine at 28 °C displayed punctate fluorescence of pyranine, indicating that the liposomes were trapped in endosome. However, after heating at 45 °C for 15 min, the same cells exhibited diffuse fluorescence of pyranine, indicating that destabilization of the liposomes in endosome with an acidic environment in combination with the brief heating caused efficient transfer of the contents into cytosol. The dual signal-responsive liposomes might have usefulness for site-specific delivery of membrane-impermeable molecules, which exhibit bioactivities in the intracellular spaces, such as siRNA and proteins.     

Development of the head and trunk mesoderm in the dogfish,Scyliorhinus torazame: II. Comparison of gene expression between the head mesoderm and somites with reference to the origin of the vertebrate head

The vertebrate mesoderm differs distinctly between the head and trunk, and the evolutionary origin of the head mesoderm remains enigmatic. Although the presence of somite‐like segmentation in the head mesoderm of model animals is generally denied at molecular developmental levels, the appearance of head cavities in elasmobranch embryos has not been explained, and the possibility that they may represent vestigial head somites once present in an amphioxus‐like ancestor has not been ruled out entirely. To examine whether the head cavities in the shark embryo exhibit any molecular signatures reminiscent of trunk somites, we isolated several developmentally key genes, including Pax1, Pax3, Pax7, Pax9, Myf5, Sonic hedgehog, and Patched2, which are involved in myogenic and chondrogenic differentiation in somites, and Pitx2, Tbx1, and Engrailed2, which are related to the patterning of the head mesoderm, from an elasmobranch species, Scyliorhinus torazame. Observation of the expression patterns of these genes revealed that most were expressed in patterns that resembled those found in amniote embryos. In addition, the head cavities did not exhibit an overt similarity to somites; that is, the similarity was no greater than that of the unsegmented head mesoderm in other vertebrates. Moreover, the shark head mesoderm showed an amniote‐like somatic/visceral distinction according to the expression of Pitx2, Tbx1, and Engrailed2. We conclude that the head cavities do not represent a manifestation of ancestral head somites; rather, they are more likely to represent a derived trait obtained in the lineage of gnathostomes.     

Treadmill running induces satellite cell activation in diabetic mice

Skeletal muscle-derived stem cells, termed as satellite cells, play essential roles in regeneration after muscle injury in adult skeletal muscle. Diabetes mellitus (DM), one of the most common metabolic diseases, causes impairments of satellite cell function. However, the studies of the countermeasures for the DM-induced dysfunction of satellite cells have been poor. Here, we investigated the effects of chronic running exercise on satellite cell activation in diabetic mice focused on the molecular mechanism including Notch and Wnt signaling, which are contribute to the fate determination of satellite cells. Male C57BL/6 mice 4 weeks of age were injected with streptozotocin and were randomly divided into runner group and control group. Runner group mice were performed treadmill running for 4 weeks. DM attenuated satellite cell activation and the expressions of the components of Notch and Wnt signaling. However, chronic running resulted in activation of satellite cells in diabetic mice and salvaged the inactivity of Wnt signaling but not Notch signaling. Our results suggest that chronic running induces satellite cell activation via upregulation of Wnt signaling in diabetic as well as normal mice.