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.     

Bilateral polynomial matrix equations in two indeterminates

Two special cases of the bilateral 2-D polynomial matrix equationDU +VN=C whenC=I andC=I with being a -stable 2-D polynomial, which are related respectively to deadbeat and asymptotic control problems of 2-D systems, are first considered. By generalizing the concepts of factor coprimeness, zero coprimeness and zero skew primeness in the 2-D polynomial ring to the ring of causal -stable 2-D rational functions, a constructive solution of these two problems mentioned is proposed. Based on these results, we derive a solvability condition for the bilateral equiation whereC is a general 2-D polynomial matrix. The general solutions are investigated as well.     

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.     

Photolytic crosslinking of poly(p-hydroxystyrene)

Light-induced crosslinking of poly(p-hydroxystyrene) (PPHS) is significantly enhanced by O₂. This was evidenced by molar mass (light scattering measurements) and by gel content determinations which were performed on various polymer samples before and after continuous irradiation at λ_inc = 254 nm. The following mechanism was elucidated with the aid of flash photolysis studies: Crosslinking in the absence or presence of O₂ is mainly due to the combination of phenoxyl type radicals. In the absence of O₂ the latter are exclusively formed by O? H bond cleavage of singlet excited phenolic groups. Triplet excited phenolic groups which are also formed do not deactivate via O? H bond cleavage but react very effectively with O₂. This reaction leads to the formation of HO and additional phenoxyl type radicals. All Commercial and most laboratory-prepared PPHS samples contain chemically bound impurities of quinoid nature. On the basis of results performed with model compounds of low molar mass, it is concluded that triplet excited quinoid groups react effectively with phenolic groups forming phenoxyl type radicals and that they are quite unreactive with respect to the abstraction of alphatic hydrogen atoms. Irradiation of PPHS at λ_inc = 254 nm causes the formation of quinoid groups which absorb strongly at this wavelength. Light absorption by these groups becomes a determining factor with respect to photochemical alteration in the course of further irradiation.     

"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.     

An experimental study of the thermodynamic properties of 1,1-difluoroethane

Experimental vapor pressures andP--T data of an important alternative refrigerant, 1, 1-difluoroethane (HFC-152a), have been measured by means of a constant-volume method coupled with expansion procedures. SixtyP--T data were measured along eight isochores in a range of temperaturesT from 330 to 440 K, at pressuresP from 1.6 to 9.3 MPa, and at densities from 51 to 811 kg·m^–3. Forty-six vapor pressures were also measured at temperatures from 320 K to the critical temperature. The uncertainties of the temperature and pressure measurements are within ±7mK and ±2kPa, respectively, while the uncertainty of the density values is within ±0.1%. The purity of the sample used is 99.9 wt%. On the basis of the measurements along each isochore, five saturation points were determined and the critical pressure was determined by correlating the vapor-pressure measurements. The second and third virial coefficients for temperatures from 360 to 440 K have also been determined.