Enhancement of adenovirus-mediated gene transfer to human bone marrow cells

Adenovirus infection of CD34+ hematopoietic stem/progenitor cells is dependent on the multiplicity of infection (MOI), time of incubation, the volume in which the co-incubation occurs and the presence or absence of growth factors. Studies revealed that a brief co-incubation (1-8 hours), resulted in low levels of transgene expression, suggesting that adenovirus infection of CD34+ cells occurs slowly, and optimal transduction requires a 24 hour exposure to adenovirus. Infection by Ad/beta-gal or Ad/p53 at a MOI of 500:1 provided a high transduction efficiency but inhibited hematopoietic function. However, treatment at a MOI of 50-100 resulted in efficient transduction (10.7-15.7% positive) without detectable toxicity. Secondary proof of adenovirus transgene expression was demonstrated by detection of mRNA for p53 in Ad/p53 infected stem cells. We conclude that a 24 hour exposure to recombinant adenovirus encoding p53 or beta-gal, at a MOI of 50-100 is optimal for in vitro gene transfer to BM cells and has no significant effect on hematopoietic function. Adenovirus-mediated transduction of BM cells can also be modulated by growth factors (IL-3, GM-CSF and G-CSF) with improved gene delivery and maintenance of hematopoietic function. In summary, adenovirus vectors can be used to transiently transduce stem cells, and conditions have been defined to maximize expression and limit inhibitory effects on CD34+ cells. These data support continued investigation of this vector for local cytokine delivery and purging of stem cell products.     

Isomorphous replacement of cystine with selenocystine in endothelin: oxidative refolding, biological and conformational properties of [Sec3,Sec11,Nle7]-endothelin-1

Air re-oxidation of fully reduced human endothelin-1 under optimized conditions yields the natural isomer with parallel disulfide bridges and the non-natural isomer with crossed disulfide bridges at a ratio of 3:1. In view of the recently determined highly reducing redox potential of selenocysteine (-381 mV) in peptides, the half-cystine residues Cys3 and Cys11 of the natural isomer of endothelin-1 were replaced by selenocysteine. Taking advantage of the high stability of the diselenide group toward reducing agents for disulfides a regioselective disulfide bridging of the second cysteine pair allowed for straightforward preparation of the [Sec3,Sec11, Nle7]-endothelin-1. NMR structural analysis showed conformational preferences of this endothelin analog that were identical to those of the natural hormone. Similarly, the bioactivity data confirmed that replacement of cysteine residues with selenocysteine was without detectable effect on receptor recognition and signal transduction. Both findings strongly support that the exchange of sulfur against selenium produces a fully isomorphous molecule as recently observed for similar exchanges at the level of methionine residues in proteins. Moreover, oxidative refolding of the fully reduced [Sec3,Sec11,Nle7]-endothelin-1 fulfilled the expectation that the redox potential of the selenocysteines would dictate quantitative formation of the natural isomer. These results suggest that the selenocysteine approach, besides offering an interesting chemical tool for induction of correct oxidative folding of multiple cysteine-containing peptides, should even allow for the preparation of non-natural isomers and thus for studying conformational preferences of folding intermediates in peptides and proteins.     

Two patients with polymyositis or dermatomyositis complicated with massive pleural effusion

Two patients with polymyositis (PM) or dermatomyositis (DM) complicated with massive pleural effusion are reported here. Both patients presented a high-grade fever, pleural effusion prominent on the right, and good response to steroid therapy. In a 50-year-old woman with PM, combined process of pleural inflammation, cardiomyopathy and coexisting hypothyroidism were considered to be responsible for the accumulation of the massive pleural effusion. However, in a 34-year-old man with DM, pleural inflammation associated with interstitial pneumonia or pleural microvasculopathy in DM was considered to be responsible for the accumulation of the massive pleural effusion.     

Diagnosis and rehabilitation strategies for patients with hysterical hemiparesis: a report of four cases

BACKGROUND: Sulphidoleukotrienes (slt) are important mediators in allergic diseases that are synthesized after allergen-specific stimulation. OBJECTIVES: The aim of this study is to determine in vitro slt production after allergen-specific (Dermatophagoides pteronyssinus) stimulus of peripheral blood leucocytes and to observe whether histamine release in whole blood with the same allergen correlates with slt production. We also wanted to evaluate whether a correlation exists between the release of slt and histamine and other diagnostic procedures as well as various clinical situations. METHODS: We studied 62 patients sensitive to Dermatophagoides pteronyssinus (Der p), 30 atopic controls and 12 healthy donors. We determined slt production using the CAST-ELISA technique and histamine release using two concentrations of Der p extract (20 and 2 ng/mL). We also carried out quantification of specific and total IgE levels, skin tests and pulmonary function test on each patient. RESULTS: We observed a significantly increased slt release after in vitro stimulation with Der p. There was a significant difference in the slt release between controls and sensitive patients (P < 0.001) and between atopic controls and sensitive patients (P < 0.001). The data are similar to those obtained with histamine release. We noted a positive correlation (P < 0.001) between slt and histamine release (r = 0.71, at 2 ng/mL and r = 0.83 at 20 ng/mL). We also found a positive (P<0.001), although weak (r=0.4 with at 2ng/mL, and r = 0.34 with P = 0.003 at 20 ng/mL) correlation between slt release and specific IgE levels as well as between slt release and skin-test reactivity (r = 0.49 at 2 ng/mL and r = 0.45 at 20 ng/mL; P < 0.001). No significant correlation between slt release and asthma severity was observed, although a trend toward higher slt production in severe and moderate asthma was detected. We found a significant (P<0.001) but weak (r=-0.3) negative correlation between age and slt release. With respect to sex-related differences, we found significant differences (P < 0.05) in slt release between the sexes with a higher slt release in men than in women. CONCLUSION: We conclude that CAST-ELISA for quantification of slt production is a useful in vitro method for diagnosing sensitization to Der p. There also exists a close correlation between slt release and other parameters of allergic sensitization in vitro as well as in vivo.     

Importance of nuclear generation in the struggles to reduce CO2 emission at Kansai Electric Power Co.

It has been pointed out in recent years that the potential impacts of global warming has been becoming more and more serious because of the rapid increase of anthropogenic CO₂ emission.

Japan's annual CO₂ emissions (fiscal 1994) amounted to 343 million tons of carbon. Although CO₂ emissions caused by fossil-fuel power generation accounted for 29.4% of total, on a sector basis, those directly from the energy conversion sector accounted for only 7.7%. Most CO₂ emissions (21.7% of total) resulted from electric power use in the industrial, commercial and domestic sectors. Thus, the reduction of CO₂ emissions caused by the use of electricity is a nationwide subject.

Understanding that both supply side and demand side approaches are necessary, Kansai Electric has been deploying “New ERA Strategy” as a comprehensive strategy to seek a potential for CO₂ reduction more broadly and deeply. Among a number of action items are the promotion of nuclear power generation, and improvement of overall energy efficiency, besides such demand side measures as leveling off the peak load.

The effectiveness of action items of the New ERA Strategy was evaluated in terms of CO₂ reduction. As a result, estimated CO₂ reduction related to nuclear power amounted to 88% of the total for fiscal 1995 in comparison with 1990, and that expected in 2000 is 84%. These results reconfirm that nuclear power is always the key to practical CO₂ reduction at present and in the future.

Comparison with candidate technology alternatives revealed that photovoltaic power generation needed 7 times greater rated capacity and 280 times larger area than nuclear power, so it is not realistic as a central power station alternative. The comparison also clarified that if wind power stations were constructed at all feasible sites in the Kansai region, they would not be a viable alternative to a single nuclear unit from CO₂ reduction viewpoint.     

Vivid image projection system using e‐Paper active screen

Projected image on a screen is not always vivid enough when it is projected in a bright room. We generally have to choose either vivid image in a dark room or dull image in a bright room. We have suggested a new projection system which can realize high contrast image projection in a bright room. It is consisted of electronic paper screen, projector, and light‐emitting diode room light. The electronic paper (e‐Paper) screen alternates its whole surface white/black with 120 Hz. Room lights are controlled with the same frequency of 120 Hz with the opposite phase as that of the e‐Paper screen. We have confirmed that our new system has achieved higher contrast than the conventional projection system and also enough readability of paper documents in the room simultaneously. We have thus confirmed that our new system can bring an ideal condition, for audience, which can realize a vivid image projection in a bright room.     

A novel quarter‐wave retardation film for improving viewing angle properties in time‐sequential stereoscopic 3D‐LCDs

We have successfully developed a quarter‐wave retardation film (QWF) for wide viewing angle 3D liquid crystal displays (3D‐LCDs) that provides high luminance, low crosstalk, low color change, and low head‐tilt‐angle dependency. It was found that the out‐of‐plane retardation (Rth) of the QWF in the LCD needs to be close to 0 nm in order to improve the 3D display properties at an off‐axis position and that the in‐plane retardation (Re) needs to be adjusted from 120 to 130 nm to achieve low color change with head tilting. We adopted a coating process for making our QWF because of its potential for retardation control. 3D‐LCDs with this QWF whose Rth was nearly zero had high performance and allowed off‐axis other than on‐axis.     

Constructive model-based analysis for safety assessment

The aerospace industry still uses fault trees to perform reliability analysis. This is because fault-tree modeling and analysis (FTA) seems easier to practical engineers when compared with Markov models, even though FTA provides a weaker form of analysis. In this paper, we propose an automatic strategy for generating Markov-based models and corresponding analysis formulations, according to ARP 4761, directly from Simulink diagrams annotated with failure information. The generated Markov-based models are expressed in the formal language PRISM, and the analysis is carried out by the PRISM model checker. The strategy is compositional and based on a comprehensive set of translation rules from Simulink to PRISM. We briefly address soundness and completeness of the rules and, to illustrate the application of the strategy, we apply it to a classical avionics case study: an actuator control system.     

"Pored-domes" of the fenestrated endotheliocyte of the glomerular and peritubular capillaries in the rodent kidney.

The fenestrated endotheliocyte of peritubular and glomerular capillaries in rat and mouse kidneys were observed with SEM and TEM. In the glomerular capillary, so-called "pored-domes" were found not only at the fenestrated areolae but also at the nuclear region of the endotheliocyte. At the region between filtration surface and nuclear region, they accumulated to construct a sponge-like structure. The endotheliocyte of peritubular capillary also showed small "pored-domes". The size and morphology of the pores in the "pored-domes" of glomerular and peritubular capillaries were similar to those of areolae fenestratae of the respective capillary. Based on the findings, we assumed that pored-domes and the sponge-like structure are the reservoir for the fenestrated area of the endotheliocyte to accommodate the rapid expansion of capillary lumen.