A novel strategy of cell targeting based on tissue-specific expression of the ecotropic retrovirus receptor gene

Gene transfer into specific tissues or cell types is a key technique in the development of gene therapy. Modification of vector particles such that they selectively bind to the target cells has been attempted, but the limitation of this approach is the low transduction efficiency. Here, we show that a two-step gene transfer system can be used for efficient cell targeting. With this strategy, and using a high-titer adenoviral vector containing a tissue-specific promoter, we have engineered a system in which only target cells become susceptible to retrovirus-mediated transduction. In a model experiment, we constructed an adenoviral vector (Ad.AFPEcoRec) containing the ecotropic retrovirus receptor (EcoRec) gene under the control of the alpha-fetoprotein (AFP) promoter. A binding assay showed that after transduction with AD.AFPEcoRec, EcoRec molecules were efficiently expressed in AFP+HepG2 cells, but not in AFP-HeLa and AFP-HLE cells. The EcoRec-expressing HepG2 cells could be stably transduced with ecotropic retroviral vectors, whereas HeLa and HLE cells remained highly resistant to retrovirus-mediated gene transfer. The apparent titer on HepG2 cells was greater than 2 x 10(5) CFU/ml. Because various tissue-specific promoter/enhancer elements are available, the two-step system could be used as a general strategy for both ex vivo and in vivo targeted gene transfer.     

Modeling of the pharyngeal muscle in Caenorhabditis elegans based on FitzHugh-Nagumo equations

The pharyngeal pumping motion to send food to the bowel is a rhythmic movement in Caenorhabditis elegans. This paper proposes a simulation-based approach to investigate the mechanisms of rhythm phenomena in the pharyngeal pumping motion. To conduct the simulations, first, we developed a pharyngeal muscle model including 29 cell models which simulate the activity of each cell as a membrane potential based on FitzHugh-Nagumo equations. Then, to compare the response of the model with that of C. elegans, we calculated the electropharyngeogram (EPG), which represents the electrophysiological responses of the pharyngeal cells, using the simulated membrane potentials. The results confirmed that our model could generate the EPG similar to that measured from C. elegans. We proposed a computer simulation of the pumping motion to investigate the mechanisms of rhythm phenomena in living organisms.     

Thickness dependence of H2 gas sensor in amorphous SnQx films prepared by ion-beam sputtering

Amorphous SnO_x films were deposited by ion-beam sputtering on sintered alumina substrates. Amorphous film sensors were prepared by annealing the films at 300° C for 2 h in air. The thickness dependence of resistivity and hydrogen gas sensitivity were measured at 150° C over the thickness range 1 to 700 nm. A resistivity maximum was observed in ultrathin films. Resistivity increased by three orders of magnitude with increasing film thickness from 0.9 to 7.4 nm and then decreased by five orders of magnitude from 7.4 to 35 nm. Ultrathin film sensors showed sensitivity maxima around a thickness of 10 nm. Sensitivity and resistivity of ultrathin films were significantly influenced by the thermal expansion coefficient and the surface state of the substrate.     

Properties of indium oxide/tin oxide multilayered films prepared by ion-beam sputtering

The preparation and characterization of indium oxide (InO _x )/tin oxide (SnO _y ) multilayered films deposited by ion-beam sputtering are described and compared with indium tin oxide (ITO) films. The structure and the optoelectrical properties of the films are studied in relation to the layered structures and the post-deposition annealing. Low-angle X-ray diffraction analysis showed that most films retained the regular layered structures even after annealing at 500° C for 16 h. As an example, we obtained a resistivity of 6×10^–4 cm and a transparency of about 85% in the visible range at a thickness of 110 nm in a multilayered film of InO _x (2.0 nm)/SnO _y (0.2 nm)×50 pairs when annealed at 300° C for 0.5 h in air. Hall coefficient measurements showed that this film had a mobility of 17 cm² V^–1 sec^–1 and a carrier concentration (electron density) of 5×10²⁰ cm^–3.     

Pressure-induced conversion of α-connectin to β-connectin

The mechanism of the pressure-induced tenderization of meat has not been fully established in spite of its beneficial effect. To detect the changes in the large structural proteins of the myofibrils induced by pressurization without heat treatment, high hydrostatic pressure (100-300 MPa) was applied to rabbit at-death skeletal muscle for 10 min at low temperature (0-2°C). Significant differences in the electrophoretic pattern of connectin (also called titin) in isolated myofibrils were observed between the control and pressurized muscle samples. The conversion of α-connectin (2800 kDa) to β-connectin (2100 kDa) was accelerated with increasing pressure applied to the muscle; also nebulin (800 kDa) was degraded by pressure treatment. From the results it is clear that the degradation of connectin is induced by pressurization alone without heat treatment. If the conversion of α-connectin to β-connectin during conditioning has some influence on meat tenderization, the pressure-induced conversion of α- to β-connectin is possibly one of the causes of pressure-induced tenderization of meat.     

Deformation of NiO single crystals below room temperature: dislocation configurations

NiO single crystals prepared by two crystal growth techniques (zone melting in an arc-image furnace and Verneuil crystallization have been deformed by compression along 001 at temperatures as low as 4.2 K, and the dislocation substructure observed by transmission electron microscopy. The Peierls mechanism has been suggested as the mechanism controlling the mechanical behaviour at the lower temperatures. The dislocations generated at cavities found in the zone-melted crystals may be responsible for the increase of the flow stress of these crystals compared with the Verneuil ones.     

Surface tension of liquid3He and4He near the liquid-gas critical point

The surface tension of liquid³He and⁴He was measured near the gas-liquid critical points in the reduced temperature range 3×10^–4–1, where t (T _c –T)/T _c . The critical exponents were found to be ₃=1.289±0.015 for³He and ₄=1.306±0.017 for⁴He. These values are very close to those for classical liquids, and are consistent with the value of 1.28 predicted by Widom, but are apparently different from the exponents previously obtained for liquid helium isotopes, which are near unity. The critical coefficients show good agreement with the quantum-corrected corresponding states theory for the Lennard-Jones 6–12 potential discussed by Young. The interface thickness is deduced from Widom's theory to bed=d ₀t^–v withd ₃₀=0.14±0.03nm and v₃=0.57±0.04 for³He, andd ₄₀=0.37±0.07 nm and v₄=0.58±0.01 for⁴He.     

Stromal CCL5 Promotes Breast Cancer Progression by Interacting with CCR3 in Tumor Cells

Chemokines secreted from stromal cells have important roles for interactions with carcinoma cells and regulating tumor progression. C-C motif chemokine ligand (CCL) 5 is expressed in various types of stromal cells and associated with tumor progression, interacting with C-C chemokine receptor (CCR) 1, 3 and 5 expressed in tumor cells. However, the expression on CCL5 and its receptors have so far not been well-examined in human breast carcinoma tissues. We therefore immunolocalized CCL5, as well as CCR1, 3 and 5, in 111 human breast carcinoma tissues and correlated them with clinicopathological characteristics. Stromal CCL5 immunoreactivity was significantly correlated with the aggressive phenotype of breast carcinomas. Importantly, this tendency was observed especially in the CCR3-positive group. Furthermore, the risk of recurrence was significantly higher in the patients with breast carcinomas positive for CCL5 and CCR3 but negative for CCR1 and CCR5, as compared with other patients. In summary, the CCL5-CCR3 axis might contribute to a worse prognosis in breast cancer patients, and these findings will contribute to a better understanding of the significance of the CCL5/CCRs axis in breast carcinoma microenvironment.     

Links between Immune Cells from the Periphery and the Brain in the Pathogenesis of Epilepsy: A Narrative Review

Accumulating evidence has demonstrated that the pathogenesis of epilepsy is linked to neuroinflammation and cerebrovascular dysfunction. Peripheral immune cell invasion into the brain, along with these responses, is implicitly involved in epilepsy. This review explored the current literature on the association between the peripheral and central nervous systems in the pathogenesis of epilepsy, and highlights novel research directions for therapeutic interventions targeting these reactions. Previous experimental and human studies have demonstrated the activation of the innate and adaptive immune responses in the brain. The time required for monocytes (responsible for innate immunity) and T cells (involved in acquired immunity) to invade the central nervous system after a seizure varies. Moreover, the time between the leakage associated with blood–brain barrier (BBB) failure and the infiltration of these cells varies. This suggests that cell infiltration is not merely a secondary disruptive event associated with BBB failure, but also a non-disruptive event facilitated by various mediators produced by the neurovascular unit consisting of neurons, perivascular astrocytes, microglia, pericytes, and endothelial cells. Moreover, genetic manipulation has enabled the differentiation between peripheral monocytes and resident microglia, which was previously considered difficult. Thus, the evidence suggests that peripheral monocytes may contribute to the pathogenesis of seizures.