Synthesis and cleavage of oligodeoxynucleotides containing a 5-hydroxyuracil residue at a defined site

Oxidation and hydrolysis of a cytosine residue can lead to the formation of 5-hydroxyuracil in DNA. The biological consequences of this modification are not fully understood. To facilitate biochemical and biophysical studies aimed at elucidating the effects of this modification in DNA, we have developed a solid-phase synthetic method for the placement of 5-hydroxyuracil residues at defined sites in oligodeoxynucleotides. This method is based upon the enhanced acidity of the 5-hydroxyl proton which allows selective aqueous acetylation. Under standard aqueous ammonia deprotection conditions, however, we observed that 5-hydroxyuracil residues are lost substantially from synthetic oligonucleotides. Substitution of aqueous ammonia with methanolic potassium carbonate and the use of phosphoramidite derivatives with alternatively protected amino groups allow synthesis of oligonucleotides containing 5-hydroxyuracil and all normal bases in high yield. The composition of the oligodeoxynucleotides prepared by this method has been verified by enzymatic digestion followed by high-performance liquid chromatography (HPLC) analysis as well as acid hydrolysis followed by GC/MS analysis. The location of the 5-hydroxyuracil residue is demonstrated by selective permanganate oxidation of the 5-hydroxyuracil residue followed by beta-elimination. We have also probed a synthetic oligonucleotide containing a unique 5-hydroxyuracil residue with uracil DNA N-glycosylase, previously reported to remove this lesion from DNA.     

Suppression of substance P biosynthesis in sensory neurons of dorsal root ganglion by prodrug esters of potent peptidylglycine alpha-amidating monooxygenase inhibitors

Substance P as well as many other neuropeptides are synthesized as glycine-extended precursors and converted to the biologically active C-terminal amides by posttranslational modification. The final step of posttranslational processing is catalyzed by peptidylglycine alpha-amidating monooxygenase (PAM). In a previous study, N-substituted homocysteine analogs were found to be potent inhibitors of PAM partially purified from conditioned medium of cultured rat medullary thyroid carcinoma CA-77 cells. These compounds, however, were only modest inhibitors of substance P production in cultured dorsal root ganglion cells, possibly because of poor cell penetration. Several ester derivatives of hydrocinnamoyl-phenylalanyl-homocysteine, one of the most potent PAM inhibitors, were prepared to increase the intracellular accessibility of these compounds. Hydrocinnamoyl-phenylalanyl-(S-benzoyl-homocysteine) benzyl ester was identified as the most potent compound, inhibiting substance P biosynthesis in dorsal root ganglion cells with an IC50 of 2 microM. Inhibition of PAM resulted in a concomitant increase in the glycine-extended substance p (substance P-Gly) precursor peptide. In the presence of 3 microM benzyl ester derivative, the intracellular substance P-Gly level was 2.4-fold higher while the substance P level was 2.1-fold lower than the corresponding peptides in control cells. These results suggest that PAM inhibition represents an effective method for suppression of substance P biosynthesis and, therefore, may have therapeutic utility in conditions associated with elevated substance P levels. Furthermore, PAM inhibition may also prove useful in decreasing other amidated peptides.     

Isolated third nerve palsy due to sarcoidosis

We present a 28-year-old man with isolated third nerve palsy as the initial manifestation of sarcoidosis. MRI demonstrated abnormal enhancement of the cisternal portion of the right third nerve. One month after steroid therapy, he showed almost complete recovery with resolution of the contrast enhancement on MRI. This patient illustrates the usefulness of enhanced MRI for detection of cranial nerve lesions of sarcoidosis.     

Oral administration of tritiated water (HTO) in mouse. III: Low dose-rate irradiation and threshold dose-rate for radiation risk

Primary vesicoureteric reflux (VUR) diagnosed on investigation of foetal hydronephrosis accounts for many antenatally detected uropathies. In order to study foetal VUR and its consequences, newborns with foetal hydronephrosis were investigated by ultrasound, micturating cystourethrogram and 99mTechnetium-dimercapto-succinic acid (DMSA), after beginning of chemoprophylaxis. Twenty-eight infants with VUR (bilateral in 15 cases) were identified giving a total 43 renal units for study. There was a predominance of males (86%), moderate/severe reflux (84%) and renal damage (51%). Presence of renal damage was correlated with the severity of reflux. VUR should be investigated in cases of foetal hydronephrosis and our results support that renal damage is frequently congenital and not secondary to urinary tract infection.     

Effect of off-orientation of seed crystal on silicon carbide (SiC) single-crystal growth on the (11 $$\bar 2$$ 0) surface0) surface

The effect of off-orientation growth has been investigated in terms of stacking fault formation during physical vapor transport (PVT) growth of silicon carbide (SiC) single crystals on the (11 0) seed crystal surface. Occurrence of stacking fault formation is largely dependent on the direction of off-orientation, and basal plane stacking fault density is significantly reduced by growing the crystals on a (11 0) seed crystal off-oriented toward 〈0001〉. The density of the basal plane stacking faults rapidly decreases from 100–150 cm⁻¹ to ∼10 cm⁻¹ as the degree of off-orientation is increased from 0 to 10 deg. The results are interpreted in the framework of microscopic facet formation during PVT growth, and the introduction of off-orientation of seed crystal is assumed to prevent (01 0) and (10 0) microfacet formation on the (11 0) growing surface through modification of the surface growth kinetics and to suppress the stacking fault formation. An erratum to this article is available at .     

A Fuzzy Behavior-Based Control for Mobile Robots Using Adaptive Fusion Units

It is known that a behavior-based control approach is effective for acquiring an intelligent control system of robots. However, further improvements are required for making any behavior-based control system robust against changes in the environments. A module learning method has been applied in the framework of fuzzy behavior-based control to have an adaptive behavioral fusion. In this paper, an adaptive fusion strategy is proposed to adaptively select a cooperative fusion unit or competitive fusion unit, depending on the external sensor information. Some simulations are given to illustrate that the present control systems are flexible against the change of environments or untrained environments, compared to those with a conventional priority-based fusion unit.     

An ultrasonic linear motor using ridge-mode traveling waves

A new type of ultrasonic linear motor is presented using traveling waves excited along a ridge atop a substrate. The ridge cross section was designed to permit only the fundamental mode to be excited during operation of the motor, with a Langevin transducer used as the source of vibration in this study. The ridge waveguide was first made of lossy media to avoid reflecting vibration energy back toward the vibration source, forming a traveling wave. A 5-mm-wide, 15-mm-tall rectangular acrylic ridge was used to move a slider placed upon it toward the vibration source, in opposition to the direction of the traveling wave transmitted along the waveguide ridge. Using a low-loss 3 x 6-mm aluminum rectangular ridge combined with a damper clamped onto the far end of the waveguide, similar results were obtained. To obtain bidirectional operation, the damper was replaced with a second Langevin transducer, giving a pair of transducers located perpendicularly to the ends of the ridge and driven with an appropriate phase difference. The moving direction of the slider was reversed by shifting this phase difference by about 180 degrees. With this simple configuration, it may soon be possible to fabricate a linear micromotor system on a silicon substrate or other semiconductor wafer adjacent to other electronic and optoelectronic devices.