Effects of Xenopus laevis mitochondrial single-stranded DNA-binding protein on primer-template binding and 3'-->5' exonuclease activity of DNA polymerase gamma

Mitochondrial DNA (mtDNA) is replicated by DNA polymerase gamma by a strand displacement mechanism involving mitochondrial single-stranded DNA-binding protein (mtSSB). mtSSB stimulates the overall rate of DNA synthesis on singly-primed M13 DNA mainly by stimulating the processivity of DNA synthesis rather than by stimulating primer recognition. We used electrophoretic mobility shift methods to study the effects of mtSSB on primer-template recognition by DNA pol gamma. Preliminary experiments showed that single mtSSB tetramers bind tightly to oligo(dT) single strands containing 32 to 48 residues. An oligonucleotide primer-template was designed with an 18-mer primer annealed to the 3'-portion of a 71-mer template containing 40 dT residues at its 5'-end as a binding site for mtSSB. DNA pol gamma bound to this primer-template either in the absence or presence of mtSSB in complexes that remained intact and enzymatically active following native gel electrophoresis. Association of mtSSB with the 5'-dT40-tail in the 18:71-mer primer-template reduced the binding of DNA polymerase gamma and the efficiency of primer extension. Binding of mtSSB to single-stranded DNA was also observed to block the action of the 3'-->5' exonuclease of DNA polymerase gamma. The size of fragments protected from 3'-->5' exonuclease trimming increases with increasing ionic strength in a manner consistent with the known salt dependence of the binding site size of Escherichia coli SSB.     

N2- and C8-substituted oligodeoxynucleotides with enhanced thrombin inhibitory activity in vitro and in vivo

2'-Deoxyguanosine (G) analogues carrying various hydrophobic substituents in the N2 and C8 positions were synthesized and introduced through solid-phase synthesis into 15-mer oligodeoxynucleotide, GGTTGGTGTGGTTGG, which forms a chairlike structure consisting of two G-tetrads and is a potent thrombin inhibitor. The effects of the substitutions at N2 and C8 of the G-tetrad-forming G residues on the thrombin inhibitory activity are relatively small, suggesting that these substitutions cause relatively small perturbations on the chairlike structure formed by the oligodeoxynucleotide. Introduction of a benzyl group into N2 of G6 and G11 and naphthylmethyl groups into N2 of G6 increased the thrombin inhibitory activity, whereas other substituents in these positions had almost no effect or decreased the activity. Particularly, the oligodeoxynucleotide carrying a 1-naphthylmethyl group in the N2 position of G6 showed an increase in activity by about 60% both in vitro and in vivo. Substitutions on the N2 position of other G residues had little effect or decreased the activity. Introduction of a relatively small group, such as methyl and propynyl, into the C8 positions of G1, G5, G10, and G14 increased the activity, presumably due to the stabilization of a chairlike structure, whereas introduction of a large substituent group, phenylethynyl, decreased the activity, probably due to the steric hindrance.     

The effective density and transport properties of compacted carbon nanotubes and nanowhiskers

We have studied the effective density, electric conductivity, and thermo emf of compacted carbon nanotubes (NTs) and nanowhiskers (NWs). The effective densities of NTs and NWs are 1.6 and 2.8 g/cm³ respectively. The thermo emf of compacted NWs (33 μV/K) and NTs (7–8μV/K) make it possible to use these materials in sensors of molecular gases and biological objects. It is also recommended to introduce NWs and NTs as fillers into glue compositions possessing increased strength and electric conductivity, which are necessary for gluing metals.     

Lightning induced disturbances in buried cables - part II: experiment and model validation

This paper presents experimental results obtained at the International Center for Lightning Research and Testing (ICLRT) at Camp Blanding, Florida during the summers of 2002 and 2003. Currents induced by triggered and natural lightning events were measured at the terminations of a buried power cable, in the cable shield, and in the inner cable conductor. Measurements of the horizontal component of the magnetic field above the ground surface for both natural and triggered lightning are also presented. For distant natural lightning events, locations of ground strike points were determined using the U.S. National Lightning Detection Network (NLDN). Based on the theoretical developments presented in Part I of this paper , the field-to-buried cable coupling equations are solved in both the time domain and in the frequency domain. The obtained experimental results are then used to validate the numerical simulations provided by the relevant developed codes.     

Calibration of scanning electron microscopes for measuring the dimensions of submicron relief elements

Slotted structures in silicon with known values of the width of the slots in the 100–500 nm range and with an error of about 1 nm were used to calibrate two model S-806 scanning electron microscopes made by Hitachi (Japan) used in different organizations. The dimensions of the electron markers of the microscopes impressed into the frame, the magnification of the microscope images, and also the values of the shifts of the maxima of the video signals relative to the edges of the elements, which must be taken into account when making measurements on actual structures, are determined. The maximum accuracy with which such measurements can be made in these microscopes is determined.Translated from Izmeritel'naya Tekhnika, No. 8, pp. 62–64, August, 1993.     

Oral health of children undergoing allogeneic bone marrow transplantation

BACKGROUND: Mixed bone marrow chimerism reliably produces donor-specific transplantation tolerance for a variety of solid organ and cellular grafts. We used a rat heterotopic tracheal transplant model for chronic rejection to investigate whether mixed chimerism could successfully prevent obstructive airway disease. METHODS: Mixed allogeneic chimeras were prepared by reconstituting lethally irradiated Wistar-Furth (WF) recipients with a mixture of 5 x 10(6) T-cell-depleted syngeneic (WF) and 100 x 10(6) T-cell-depleted allogeneic (ACI) bone marrow cells (ACI + WF --> WF). Mixed chimerism was present in all animals 28 days after bone marrow transplantation. Donor-specific, syngeneic, or major histocompatibility complex (MHC)-disparate allogeneic tracheas were implanted in recipient's omentum and removed for histologic analysis 30 to 150 days after transplantation. RESULTS: At 30 days after implantation, median luminal obstruction grades (0=none, 4=complete) of syngeneic and allogeneic tracheas were 0 and 4, respectively. Donor-specific (ACI) tracheas implanted in chimeric (ACI + WF --> WF) recipients were remarkably free of obstruction (median luminal obstruction grade=0 at 150 days) and had excellent preservation of respiratory epithelium. Third-party F344 tracheas implanted in chimeric recipients developed progressive luminal obstruction (grade 2 at 30 days, grade 3 at 90 days). CONCLUSIONS: Mixed allogeneic chimerism induces donor-specific tolerance and prevents development of the characteristic fibroproliferative obstructive lesion of bronchiolitis obliterans in a rat heterotopic tracheal transplant model. Excellent preservation of tracheal structure and morphology was achieved across major and minor histocompatibility barriers.