Accumulation of ochratoxin A in rat kidney in vivo and in cultivated renal epithelial cells in vitro

Biological applications of triplex forming oligonucleotides will require the development of oligomers with high avidity and specificity. We examined the binding enhancement resulting from intercalator conjugation to both parallel design (polythymidine T15) and antiparallel design (polypurine AG15, for binding a 15 base pair polypurine-polypyrimidine sequence in the IL-2R alpha gene enhancer) oligomers under various ionic strength and temperature conditions. Oligonucleotides were conjugated through a urea link to 6,9 diamino-3-methoxy acridine (to give T15C and AG15C). Intercalator conjugation dramatically enhanced the specific triplex binding avidity (Kd = 5 nM for AG15C and 275 nM for T15C at 25 degrees C, compared to 2 microM for AG15 and > 50 microM for T15 at 25 degrees C), without detectable binding to an inappropriate target sequence. Surprisingly, triplex formation with AG15C occurred at lower Mg2+ concentrations than with T15C. AG15 and AG15C showed rapid Mg2+ dependent self association, but not T15C or T15. T15C triplex formation occurred rapidly (completion in less than 4 min), while AG15C bound to its target sequence more slowly over 20-24 h. Thus, binding constants in the low nanomolar range are now achievable with intercalator conjugated polypurine antiparallel binding oligonucleotides, a prerequisite for biological applications of such agents.     

A note on estimating false alarm rates via importance sampling

When the statistics of the noise are non-Gaussian, analytic expressions for the probability of false alarms in detection systems are rarely available. Monte Carlo estimation techniques are therefore typically necessary. The author presents an importance sampling biasing distribution which renders exponential savings over standard Monte Carlo simulations. Two important features of this biasing strategy are that no importance sampling parameters need to be determined and no additional computations are required for implementation     

Dynamic effects in hot-carrier degradation relevant for CMOS operation

We describe different dynamic degradation effects in n- and p-MOSFETs as they are clearly proven and generally accepted to date. It turns out that they are connected with time constants in the oxide and at the interface and that time constants related to the device operation are too short to be relevant in this context. The effects are detrapping of fixed charges, the slow movement of holes in the oxide, an enhanced-degradation effect caused by alternating voltage conditions, during dynamic stress, and a post-stress interface state formation effect in nitride passivated n-MOSFETs. Furthermore, we discuss the relevance of those effects, under different operation conditions, finding that the fast non-stationary effects are of little significance. Only the “slow” effects, with time constants of seconds and above, play a role in reliability issues of MOSFETs.     

Measurement and prediction of pressure drop in two-phase flow

Experimental data for air–water two-phase co-current flow in two different pipe diameters were used to test the prediction of pressure drop by a number of existing theories and correlations. Several models are shown to be useful for prediction, particularly with the stratified regimes which have proved difficult to handle in the past. The model suggested by Olujic proved to be of particular value.     

Effects of single doses of irradiation on the expression of resistance-related proteins in murine NIH 3T3 and human lung carcinoma cells

In this report the effects of single doses of ionizing radiation on the mRNA expression of several proteins involved in multiple drug resistance were analyzed. Murine NIH 3T3 cells treated with single doses of 5, 10 and 20 Gy during the time interval from 1.5 to 72 h after irradiation were compared with their corresponding controls at the same points of time. The glutathione S-transferase-pi (GST pi) level was elevated in cells treated with 10 or 20 Gy from 24 to 72 h after irradiation compared with the control. Topoisomerase II alpha and thymidylate synthase were decreased in irradiated cells 24-72 h after exposure. These down-regulations were associated with cellular proliferation, determined by mRNA expression of the proliferation marker histone 3. Irradiated cells exhibited no alteration in the P-glycoprotein or glutathione peroxidase mRNA content. The finding that GST pi mRNA was overexpressed after irradiation was validated by investigations on a human lung carcinoma cell line (LXF 289) on the mRNA and protein level. Thus, our results indicate that irradiation alters the expression of proteins involved in multidrug resistance and may, therefore, play a role in clinical drug response.     

c-KIT expression enhances the leukemogenic potential of 32D cells

The growth of human leukemic cells in culture and in vivo is dependent upon the presence of hematopoietic growth factors. Most populations of human leukemic acute myeloblastic leukemia (AML) cells express c-Kit on their surface and respond to Kit ligand (KL) in culture. To determine if this interaction was of potential significance in vivo we used a mouse model system. 32D cells, a murine IL-3-dependent myeloid cell line, were rendered KL responsive by transfection of the murine c-Kit. After injection of 32D or 32D-Kit cells into syngeneic hosts, animals bearing 32D-Kit cells, but not 32D cells, became moribund and were killed. These animals had circulating leukemic blast cells, infiltration of bone marrow, spleen, brain, liver, lung, and kidney. Cells recovered from some of the animals continued to be dependent upon IL-3 or KL for growth while in other cases the cells were factor independent. This model illustrates that the constitutive expression of c-Kit enhances the leukemic potential of 32D cells. The model will be useful for studying the progression of leukemia in vivo and testing whether interruption of the interaction of Kit and KL can affect the growth of leukemic cells.     

Isothermic growth of one spherulite around circular obstacles in a polypropylene foil

Growth of one spherulite within a thin foil of polypropylene around one circular obstacle or around combinations of circular obstacles is investigated. For each obstacle there exists a region of shadow, seen from the nucleus of the spherulite, which influences the growth of the spherulite. Within any region of shadow the growth fronts are evolvents of the obstacle's boundary, because the spherulite grows isotropically. When two growth fronts belonging to one spherulite meet each other inside the shadow, an intrinsic grain boundary is formed for each obstacle. Additionally, growth of one spherulite around a rectangular obstacle and a spherical obstacle is investigated.     

Structural studies of TiN/ZrN multilayer coating deposited by physical vapour deposition

Multilayer coatings have been produced using unbalanced magnetron physical vapour deposition in which discrete layers of TiN and ZrN were co-deposited. It has been shown that the multilayers exhibited two-fold periodicity having values of 13 and 2.8 nm. This periodicity, measured both by cross-sectional transmission electron microscopy and low-angle X-ray diffraction, has been precisely related to the process parameters used. Analysis has shown that a third, previously unreported, phase existed within the coating having a lattice parameter comparable to that of TiN and a well-defined orientation relationship with the primary phases. The existence of this phase has been explained in terms of variations in the stoichometry of the coating due to non-uniform metal to nitrogen deposition rates within the chamber. Textural studies revealed that preferred orientation existed in the coating; the degree of which was greater in the ZrN layers than the TiN layers.     

Growth of Fe on Si (100) at room temperature and formation of iron silicide

Low-energy electron diffraction (LEED), Auger electron spectroscopy and X-ray photoelectron spectroscopy (XPS) investigations of both the growth of an iron film on silicon (100) at room temperature and the subsequent formation of iron silicide are the subjects of this paper. An in-situ cleaned silicon (100) wafer without carbon or oxygen contamination exhibiting the known 2 × 1 reconstruction in the LEED pattern served as the substrate. Iron was deposited on this reconstructed surface at 300 K. The comparison of theoretical calculations based on three growth mechanisms with XPS data obtained with take-off angles of 0° and 50° clearly demonstrates a layer-by-layer growth of the iron film on silicon (100). At 300 K no formation of iron silicide was observed, although an interaction between iron and silicon could be detected at the interface. The formation of iron silicide was observed at annealing temperatures of 630–730 K. Quantitative XPS analysis yields the presence of FeSi₂, when the thickness is large enough. Neither the iron film on silicon nor the silicide shows any LEED pattern.