DNA binding properties of human Cdc45 suggest a function as molecular wedge for DNA unwinding

The cell division cycle protein 45 (Cdc45) represents an essential replication factor that, together with the Mcm2-7 complex and the four subunits of GINS, forms the replicative DNA helicase in eukaryotes. Recombinant human Cdc45 (hCdc45) was structurally characterized and its DNA-binding properties were determined. Synchrotron radiation circular dichroism spectroscopy, dynamic light scattering, small-angle X-ray scattering and atomic force microscopy revealed that hCdc45 exists as an alpha-helical monomer and possesses a structure similar to its bacterial homolog RecJ. hCdc45 bound long (113-mer or 80-mer) single-stranded DNA fragments with a higher affinity than shorter ones (34-mer). hCdc45 displayed a preference for 3' protruding strands and bound tightly to single-strand/double-strand DNA junctions, such as those presented by Y-shaped DNA, bubbles and displacement loops, all of which appear transiently during the initiation of DNA replication. Collectively, our findings suggest that hCdc45 not only binds to but also slides on DNA with a 3'-5' polarity and, thereby acts as a molecular 'wedge' to initiate DNA strand displacement

Impact of Air Jet Impingement Technology on the Strength of Tissue Paper

Impinging air jets can be used to dewater, heat, and dry the web of tissue paper. High velocities of the air jets degrade the paper, and appropriate adjustments to the jet velocity and the distance of the nozzle from the surface of the wet web are crucial to obtain the highest quality product. This work investigated the correlation between the velocity of the air jet and the strength of paper subjected to the impingement method. Papers with an initial moisture content of 20% and various pulp mixes were tested, and the physical properties of papers were explored. After impinging an air jet, different tensile strength limits were obtained in the machine and cross directions. The paper had lower apparent density and higher roughness compared to classical pressing. The dependence of tensile strength and roughness on the fibers composition also was determined. Increasing the amount of eucalyptus fibers in impingement dewatered paper resulted in a decrease in its tensile strength and roughness. The value of elongation before breaking was the highest for softwood papers after the impingement method. The maximum velocity of an air jet that can be used to dewater or dry paper without the risk of damage to the papers was determined

Cdc45-induced loading of human RPA onto single-stranded DNA

Abstract Cell division cycle protein 45 (Cdc45) is an essential component of the eukaryotic replicative DNA helicase. We found that human Cdc45 forms a complex with the single-stranded DNA (ssDNA) binding protein RPA. Moreover, it actively loads RPA onto nascent ssDNA. Pull-down assays and surface plasmon resonance studies revealed that Cdc45-bound RPA complexed with ssDNA in the 8–10 nucleotide binding mode, but dissociated when RPA covered a 30-mer. Real-time analysis of RPA-ssDNA binding demonstrated that Cdc45 catalytically loaded RPA onto ssDNA. This placement reaction required physical contacts of Cdc45 with the RPA70A subdomain. Our results imply that Cdc45 controlled stabilization of the 8-nt RPA binding mode, the subsequent RPA transition into 30-mer mode and facilitated an ordered binding to ssDNA. We propose that a Cdc45-mediated loading guarantees a seamless deposition of RPA on newly emerging ssDNA at the nascent replication fork

Reaction of Silyl Hydrides with Tetrabutoxygermanium in the Presence of B(C₆F₅)₃: Difference between Silicon and Germanium Chemistries and Easy Route to GeH₄

Dehydrocarbonative condensation reaction between alkoxysilanes and hydrosilanes catalyzed by tris­(pentafluorophenyl)­borane has been widely used for the formation of siloxane bonds. Our attempt to extend this chemistry to the preparation of Ge–O–Si bonds produced an unexpected outcome. The reaction of Ge­(OBu)₄ with PhMe₂SiH in the presence of a catalytic amount of B­(C₆F₅)₃ at room temperature proceeded smoothly with the complete consumption of reactants and formation of GeH₄ and PhMe₂SiOBu in high yields. For the first time we have achieved selective exchange of functional groups between Ge–OR and Si–H. The discovered reaction features simple reaction conditions and can be used to prepare GeH₄ in situ from easily available and safe substrates

Dietary Intake at 9 Years and Subsequent Body Mass Index in Adolescent Boys and Girls: A Study of Monozygotic Twin Pairs

There is a lack of evidence pointing to specific dietary elements related to weight gain and obesity prevention in childhood and adulthood. Dietary intake and obesity are both inherited and culturally transmitted, but most prospective studies on the association between diet and weight status do not take genetics into consideration. The objective of this study was to document the association between dietary intake at 9 years and subsequent Body Mass Index (BMI) in adolescent monozygotic boy and girl twin pairs. This research used data from 152 twin pairs. Dietary data were collected from two 24-hour-recall interviews with a parent and the child aged 9 years. Height and weight were obtained when the twins were aged 9, 12, 13, and 14 years. Intrapair variability analysis was performed to identify dietary elements related to BMI changes in subsequent years. BMI-discordant monozygotic twin pairs were also identified to analyze the dietary constituents that may have generated the discordance. After eliminating potential confounding genetic factors, pre-adolescent boys who ate fewer grain products and fruit and consumed more high-fat meat and milk had higher BMIs during adolescence; pre-adolescent girls who consumed more grain products and high-fat meat and milk had higher BMIs during adolescence. Energy intake (EI) at 9 years was not related to BMI in subsequent years. Our study suggests that messages and interventions directed at obesity prevention could take advantage of sex-specific designs and, eventually, genetic information.Peer reviewe