Cleavage of single strand RNA adjacent to RNA-DNA duplex regions by Escherichia coli RNase H1

RNase H1 from Escherichia coli cleaves single strand RNA extending 3' from an RNA-DNA duplex. Substrates consisting of a 25-mer RNA annealed to complementary DNA ranging in length from 9-17 nucleotides were designed to create overhanging single strand RNA regions extending 5' and 3' from the RNA-DNA duplex. Digestion of single strand RNA was observed exclusively within the 3' overhang region and not the 5' overhang region. RNase H digestion of the 3' overhang region resulted in digestion products with 5'-phosphate and 3'-hydroxyl termini. The number of single strand RNA residues cleaved by RNase H is influenced by the sequence of the single strand RNA immediately adjacent to the RNA-DNA duplex and appears to be a function of the stacking properties of the RNA residues adjacent to the RNA-DNA duplex. RNase H digestion of the 3' overhang region was not observed for a substrate that contained a 2'-methoxy antisense strand. The introduction of 3 deoxynucleotides at the 5' terminus of the 2'-methoxy antisense oligonucleotide resulted in cleavage. These results offer additional insights into the binding directionality of RNase H with respect to the heteroduplex substrate.     

Is it possible to predict limb viability in complex Gustilo IIIB and IIIC tibial fractures? A comparison of two predictive indices

The patient with severe lower limb trauma presents a management dilemma; whether to amputate primarily or to attempt limb salvage. In recent years, many predictive indices have been published which purport to identify limbs which are non-viable. We retrospectively applied two recently described indices, the Mangled Extremity Severity Score (MESS) and the Limb Salvage Index (LSI), to 54 limbs in 50 patients with either Gustilo IIIB or IIIC complex tibial fractures. There were 22 amputations (40.7 per cent) in the series. The mean MESS score in the limb salvage group was 3.8 (range 2-10), and the mean MESS score in the amputation group was 7.7 (range 4-13) (P < 0.0001). The mean LSI score in the limb salvage group was 3.6 (range 3-8), and the mean LSI score in the amputation group was 6.9 (P < 0.01). However, in the group with MESS scores > 7 (which recommends amputation), there were three limbs which were salvaged with acceptable functional outcome. Similarly, in those with LSI scores > 6 (which recommends amputation), there were seven limbs successfully salvaged. A MESS > 7 offered a greater relative risk of amputation (9.2) than a LSI score > 6 (5.3). We found both indices of use in predicting limb salvage and functional outcome. However, neither is sufficiently accurate to be considered absolutely reliable in clinical practice.     

A strategy for sampling on a sphere applied to 3D selective RF pulse design

Conventional constant angular velocity sampling of the surface of a sphere results in a higher sampling density near the two poles relative to the equatorial region. More samples, and hence longer sampling time, are required to achieve a given sampling density in the equatorial region when compared with uniform sampling. This paper presents a simple expression for a continuous sample path through a nearly uniform distribution of points on the surface of a sphere. Sampling of concentric spherical shells in k-space with the new strategy is used to design 3D selective inversion and spin-echo pulses. These new 3D selective pulses have been implemented and verified experimentally.     

Fusarium fungaemia in severely neutropenic patients

We report two cases of Fusarium infection with evidence of fungaemia in severely neutropenic patients with leukaemia. One patient was a 65-year-old woman with chronic lymphocytic leukaemia infected by Fusarium verticillioides. The other patient was a 45-year-old woman with acute myeloblastic leukaemia infected by Fusarium spp. Fungaemia was the only evident manifestation of these fungal infections.     

Morphological organization of the globus pallidus-subthalamic nucleus system studied in organotypic cultures

The morphological organization of the globus pallidus (GP), the subthalamic nucleus (STN), and the pallidosubthalamic projection was studied in organotypic cultures. Coronal slices from the GP, the STN, the striatum (CPu), and the cortex (Cx) were taken from the rat after postnatal days 0-2 and grown for 2 or 5-6 weeks. For analysis, immunocytochemistry against glutamate (GLU), parvalbumin (PV), and calretinin (CR) was combined with confocal microscopy. After 2 weeks in vitro, the STN showed a densely packed, homogeneous GLU-immunoreactive (ir) cell population. Pallidal GLU-ir neurons were heterogeneous, consisting of large-sized weakly GLU-ir neurons and small-sized intensively GLU-ir neurons. After 5-6 weeks in vitro, pallidal axons had radiated from numerous large-sized PV-ir cells and selectively innervated the STN, where they heavily ramified. Cultured STN neurons were not stained for PV; however, multipolar intensely PV-ir neurons were located at the border of the STN with their dendrites oriented towards the STN. Double labeling for PV and CR in both mature cultures and in the adult rat revealed that the culture CR-ir neurons from the GP, the Cpu, and from areas adjacent to the STN were different from cultured PV-ir neurons and their morphologies and distribution corresponded to that in vivo. These results demonstrate that 1) cultured CP and STN neurons display similar morphologies found in in vivo, 2) PV-ir pallidal neurons heavily and selectively innervate the STN; 3) there is a specific class of STN border neurons; and 4) in contrast to the in vivo situation, most cultured STN neurons are PV-negative.     

A 16-Mb Toggle MRAM With Burst Modes

This paper describes a recently developed 16-Mb toggle magnetic random access memory (MRAM). It has 100-MHz burst modes that are compatible with a pseudo-SRAM even though the toggle cell requires reading and comparing sequences in write modes. To accelerate operating clock frequency, we propose a distributed-driver wide-swing current-mirror scheme, an interleaved and pipelined memory-array group activation scheme, and a noise-insulation switch scheme. These circuit schemes compensate the toggle cell timing overhead in write modes and maintain write-current precision that is essential for the wide operational margin of MRAMs. Because toggle cells are very resistant to write disturbance errors, we designed the 16-Mb MRAM to include a toggle MRAM cell. The MRAM was fabricated with 0.13-mum CMOS and 0.24-mum MRAM processes with five metal layers.     

Influence of interstripe gain on crosscoupled-mode operation intwin-stripe lasers

Cross-coupled-mode operation in a twin-stripe laser is analyzed with particular concern for interstripe gain. When the interstripe gain of a twin-stripe laser is high, cross-coupled-mode operation is obtained with a cavity longer than the coupling length of the twin-stripe waveguide. The far-field pattern is single peaked and deflected to the low carrier density side. These characteristics are in contrast to those of the previous model without interstripe gain