Formation of a DNA loop at the replication fork generated by bacteriophage T7 replication proteins

Intermediates in the replication of circular and linear M13 double-stranded DNA by bacteriophage T7 proteins have been examined by electron microscopy. Synthesis generated double-stranded DNA molecules containing a single replication fork with a linear duplex tail. A complex presumably consisting of T7 DNA polymerase and gene 4 helicase/primase molecules was present at the fork together with a variable amount of single-stranded DNA sequestered by gene 2.5 single-stranded DNA binding protein. Analysis of the length distribution of Okazaki fragments formed at different helicase/primase concentrations was consistent with coupling of leading and lagging strand replication. Fifteen to forty percent of the templates engaged in replication have a DNA loop at the replication fork. The loops are fully double-stranded with an average length of approximately 1 kilobase. Labeling with biotinylated dCTP showed that the loops consist of newly synthesized DNA, and synchronization experiments using a linear template with a G-less cassette demonstrated that the loops are formed by active displacement of the lagging strand. A long standing feature of models for coupled leading/lagging strand replication has been the presence of a DNA loop at the replication fork. This study provides the first direct demonstration of such loops.     

Synthesis of a cyclic peptide containing norlanthionine: effect of the thioether bridge on peptide conformation

Two diastereomeric analogues of ring C of nisin incorporating a novel norlanthionine residue have been synthesized via a triply orthogonal protecting group strategy. A full structural study was carried out by NMR, which elucidated the conformational properties of the two peptides and enabled the identity of each diastereoisomer to be proposed.     

Dialysis neutropenia: the role of the cytoskeleton

Dialysis neutropenia is the result of pulmonary sequestration of neutrophils after complement activation by the dialyzer membrane. Increased expression of neutrophil adhesion receptors, such as CD11b/CD18, suggests that neutrophil adhesion to the capillary endothelium is a possible mechanism. An alternative hypothesis is that the complement fragment C5a modulates neutrophil mechanical properties via the cytoskeleton-largely filamentous actin (F-actin)-stiffening them and thereby slowing their passage through the pulmonary capillaries. To investigate this hypothesis, we developed an assay to measure the F-actin content of neutrophils in whole blood using flow cytometry and the stain NBD-phallacidin. We measured neutrophil F-actin content during hemodialysis of patients with polysulfone (N = 6), Hemophan (N = 6), and Cuprophan membranes sterilized with either ethylene oxide (N = 5) or steam (N = 6). Cell counts, neutrophil and monocyte CD11b expression and plasma C5a concentrations were also measured. The results confirm the strong relationship between the degree of neutropenia, increases in CD11b expression and plasma C5a levels reported by previous researchers. Modulation of the F-actin content of neutrophils was also strongly related to C5a levels, indicating that the neutrophil cytoskeleton is active during dialysis. Modeling of cell counts suggests that with Cuprophan a substantial fraction of neutrophils and monocytes are sequestered before they even pass through the dialyzer, suggesting some form of systemic activation of these cells. Evidence for systemic activation was also seen in measurements of F-actin content, but not CD11b expression, a finding that strengthens the case for the involvement of the cytoskeleton in dialysis neutropenia.     

Shear Properties of Thin Polymeric Films

When a thin solid organic coating is interposed between two contacting and sliding surfaces we may define a strength property τ as the frictional force per unit area of solid-solid contact. This paper reviews recent work on the influence of contact pressure, temperature and sliding velocity on τ for a range of high molecular weight organic polymers and lower molecular weight organic solids. It is shown that the shear properties of these thin films resemble those of the corresponding bulk polymers if allowance is made for the high degree of molecular orientation produced in the film during sliding.     

Effects of text genre and verbal ability on adult age differences in sensitivity to test structure.

The present study examined the effects of verbal ability and text genre on adult age differences in sensitivity to the semantic structure of prose. Young and older adults of low or high verbal ability heard narrative and expository passages at different presentation rates. The results demonstrated that older adults recalled less than younger adults and that age differences in recall were larger for low-verbal adults and expository texts. However, subjects from all groups favored the main ideas in their recalls for both types of passages. The results indicate that adult age similarities in the ability to focus on the main ideas when processing prose was not compromised by the verbal ability of the subjects or the organization of the passages used. However, the results also demonstrate how the characteristics of the learner and the characteristics of the text modulate the size of the age differences observed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The role of lubricants in machining

Friction between the rake face of a tool and the freshly formed chip surface plays a vital role in influencing the ease of the metal cutting process. The existence of clean surfaces and high hydrostatic stresses favours the formation of strong adhesive friction junctions; the extent of these can be limited by the provision of a suitable lubricant. Experiments using a novel planing machine, which enables the orthogonal cutting operation to be carried out in carefully controlled atmospheres, are described; it is suggested that processes of molecular transport within a network of interfacial capillaries are dominant in controlling the effectiveness of cutting fluids provided they are capable of reaction with, or adsorption by, the metal surfaces.