Artefactual cleavage of E coli H-NS by OmpT

In the bacterium Escherichia coli, H-NS-(H1, H1a) is a heat-stable protein with a molecular mass of 15.5 kDa involved in nucleoid organisation and gene regulation linked to certain signal transduction pathways. We have shown that, following addition of preparations of everted inner membrane vesicles, heat-stable cleavage products of approximately 10 kDa of H-NS are formed in vitro from newly synthesised, radio-labelled H-NS and from purified H-NS. The 15.5 kDa protein and its cleavage products were also recovered from a minicell system. These results raised the possibility that cleavage of H-NS is physiologically significant. However, the cleavage of H-NS observed appears to occur during cell breakage and to depend on the method of protein extraction and the presence of the outer membrane protease, OmpT. Nevertheless, the results indicate that H-NS may contain at least two separate domains with cleavage occurring between these domains at a preferred OmpT site. Failure to take account of H-NS cleavage in sample preparation and analysis can lead to serious underestimation of H-NS levels.     

Study of the nutrient distribution in root tips of the tropical forage Brachiaria

A Brachiaria breeding project is being conducted at CIAT, Colombia to combine acid soil adaptation of B. decumbens with other favourable traits of Brachiaria species that are less adapted to acid soils. Micro-PIXE was applied to investigate nutrient uptake and distribution in root tips of different species grown in hydroponic culture under control and simulated acid soil stress conditions. Different sample measurement approaches were evaluated, including: (1) linear scans with single point measurements along the root axis; (2) mapping of whole root tips; and (3) mapping of root cross sections. Different tissue types could be distinguished on the base of differences in nutrient concentrations and/or Al stress. Al, if supplied under nutrient stress conditions, increased P accumulation in the central vascular tissue of the meristematic and elongation zone of B. decumbens and B. brizantha. Furthermore, a negative correlation was found between Al and Cl accumulation in the root cap of B. decumbens.     

Intrathecal fentanyl with small-dose dilute bupivacaine: better anesthesia without prolonging recovery

Recent concern regarding lidocaine neurotoxicity has prompted efforts to find alternatives to lidocaine spinal anesthesia. Small-dose dilute bupivacaine spinal anesthesia yields a comparably rapid recovery profile but may provide insufficient anesthesia. By exploiting the synergism between intrathecal opioids and local anesthetics, it may be possible to augment the spinal anesthesia without prolonging recovery. Fifty patients undergoing ambulatory surgical arthroscopy were randomized into two groups receiving spinal anesthesia with 3 ml 0.17% bupivacaine in 2.66% dextrose without (Group I) or with (Group II) the addition of 10 microg fentanyl. Median block levels reached T7 and T8, respectively (P = not significant [NS]). Mean times to two-segment regression, S2 regression, time out of bed, time to urination, and time to discharge were 53 vs 67 min (P < 0.01), 120 vs 146 min (P < 0.05), 146 vs 163 min (P = NS), 169 vs 177 min (P = NS), and 187 vs 195 min (P = NS) respectively. Motor blockade was similar between groups, but sensory blockade was significantly more intense in Group II (P < 0.01). Six of 25 blocks failed in Group I, whereas none failed in Group II. The addition of 10 microg fentanyl to spinal anesthesia with dilute small-dose bupivacaine intensifies and increases the duration of sensory blockade without increasing the intensity of motor blockade or prolonging recovery to micturition or street fitness. IMPLICATIONS: Concerns about the neurotoxicity of lidocaine have prompted efforts to find alternatives to lidocaine spinal anesthesia. We studied 50 patients undergoing ambulatory surgical arthroscopy and found that although small-dose bupivacaine alone is inadequate for this procedure, the addition of fentanyl makes it reliable.     

Hemorrhagic fever with renal syndrome

The authors describe the illness of a Czech soldier who while serving in the UNPROFOR forces in an area with the endemic occurrence of hantaviruses became infected with this agent and developed haemorrhagic fever with renal syndrome. The severe clinical course which called for seven haemodialyzation procedures led to the patient's recovery. The authors describe the course of the disease and epidemiological circumstances.     

The predominant contribution of oligopeptide transporter PepT1 to intestinal absorption of beta-lactam antibiotics in the rat small intestine

Although recent evidence suggests that certain beta-lactam antibiotics are absorbed via a specific transport mechanism, its nature is unclear. To confirm whether peptide transport in the rat can be largely ascribed to the intestinal oligopeptide transporter PepT1, the transporter has been functionally characterized and its significance in the intestinal absorption of beta-lactam antibiotics was evaluated. For evaluation of transport activity complementary RNA (cRNA) of rat PepT1 was synthesized in-vitro and expressed in Xenopus laevis oocytes. cRNA induced uptake of several beta-lactam antibiotics and the dipeptide [14C]glycylsarcosine; this was specifically inhibited by various dipeptides and tripeptides but not by their constituent amino acids or by tetra- or pentapeptides. The transport activity of PepT1 for beta-lactam antibiotics correlated well with their in-vivo intestinal transport and absorption. Furthermore, mutual inhibitory effects on uptake were observed between glyclsarcosine and beta-lactam antibiotics. Hybrid depletion of the functional expression of rat PepT1 in oocytes injected with rat intestinal epithelial total mRNA was studied using an antisense oligonucleotide corresponding to the 5'-coding region of PepT1. In oocytes injected with rat mRNA pre-hybridized with the antisense oligonucleotide against rat PepT1, the uptake of [14C]glycylsarcosine was almost completely abolished, whereas its uptake was not influenced by a sense oligonucleotide for the same region of PepT1. Similarly, the uptake of beta-lactam antibiotics was also reduced by the antisense oligonucleotide against rat PepT1. These results demonstrate that the intestinal proton-coupled oligopeptide transporter PepT1 plays a predominant role in the carrier-mediated intestinal absorption of beta-lactam antibiotics and native oligopeptides in the rat.     

High-resolution HLA-DRB typing using denaturing gradient gel electrophoresis and direct sequencing

Skeletal development of transgenic mice with a type II collagen mutation was analyzed and compared with wild-type littermates. The single base substitution in Col2a1 resulted in a glycine to serine mutation within the helical domain and corresponded to one previously identified in a patient with the lethal human chondrodysplasia, hypochondrogenesis (Horton et al. [1992] Proc. Natl. Acad. Sci. U.S.A. 89:4583-4587). Skeletal staining of embryos from 14.5 through 18.5 days of gestation demonstrated a dwarf phenotype in the transgenic embryos, most notably short limb bones and vertebral column that was first detected at 15.5 days post-coitus. In addition to the reduced length, the extent of ossification was less in the transgenic mice. The architecture of the long bone growth plate was abnormal in the transgenic tissue, in particular there was no discernible proliferative zone. There were few stacks of characteristically flattened cells and the overall length of the growth plate in the mutant embryos was reduced. At the ultrastructural level, there were fewer collagen fibrils present in the transgenic mouse cartilage compared to that of wild-type littermates. Ultrastructural localization of collagen types II, IX and XI revealed a similar pattern between the transgenic and wild-type pups, suggesting that the collagen fibrils present in the matrix of littermates with both phenotypes had a similar composition. Skeletal analysis and cartilage histochemistry indicated that effect of the type II collagen mutation was to reduce the density of the collagen fibrils within the cartilage matrix which was associated with delayed bone formation and resulted in a short-limbed phenotype.     

Mathematical foundations for error estimation in numericalsolutions of integral equations in electromagnetics

The problem of error estimation in the numerical solution of integral equations that arise in electromagnetics is addressed. The direct method (Green's theorem or field approach) and the indirect method (layer ansatz or source approach) lead to well-known integral equations both of the first kind [electric field integral equations (EFIE)] and the second kind [magnetic field integral equations (MFIE)]. These equations are analyzed systematically in terms of the mapping properties of the integral operators. It is shown how the assumption that field quantities have finite energy leads naturally to describing the mapping properties in appropriate Sobolev spaces. These function spaces are demystified through simple examples which also are used to demonstrate the importance of knowing in which space the given data lives and in which space the solution should be sought. It is further shown how the method of moments (or Galerkin method) is formulated in these function spaces and how residual error can be used to estimate actual error in these spaces. The condition number of all of the impedance matrices that result from discretizing the integral equations, including first kind equations, is shown to be bounded when the elements are computed appropriately. Finally, the consequences of carrying out all computations in the space of square integrable functions, a particularly friendly Sobolev space, are explained     

Bias and temperature dependence of homogeneous hot-electroninjection from silicon into silicon dioxide at low voltages

In this paper, new homogeneous hot-electron injection data at 300 K and 77 K is provided covering applied voltages from well below to well above the Si-SiO₂ barrier height, and a wide range of oxide fields. We found that, in contrast to the MOSFET case, homogeneous injection shows two different regimes for accelerating voltages below and above the barrier height. A simple interpretation of the data is proposed, and supported by Monte Carlo (MC) simulations of the injection experiment. Essentially, the two regimes are the signature of a marked transition between an electron population mostly heated by the electric field, and a tail population created by additional but less efficient energy gain mechanisms, leading to a sharp transition in the carrier distribution function. The details of the bias and temperature dependence of injection are then interpreted as the combined effect of tunneling and carrier distribution. Furthermore, possible implications on MOSFET gate currents are briefly discussed     

A construction of codes with exponential error bounds on arbitrarydiscrete memoryless channels

This correspondence proposes an explicit construction of codes achieving capacity for arbitrary discrete memoryless channels. The proposed code is obtained by concatenating variable inner codes and an algebraic geometry code. Further, we clarify that the proposed code achieves the error exponent obtained by Forney for concatenated codes