Structural and mechanistic insights into the DNA glycosylase AAG-mediated base excision in nucleosome.

The engagement of a DNA glycosylase with a damaged DNA base marks the initiation of base excision repair. Nucleosome-based packaging of eukaryotic genome obstructs DNA accessibility, and how DNA glycosylases locate the substrate site on nucleosomes is currently unclear. Here, we report cryo-electron microscopy structures of nucleosomes bearing a deoxyinosine (DI) in various geometric positions and structures of them in complex with the DNA glycosylase AAG. The apo nucleosome structures show that the presence of a DI alone perturbs nucleosomal DNA globally, leading to a general weakening of the interface between DNA and the histone core and greater flexibility for the exit/entry of the nucleosomal DNA. AAG makes use of this nucleosomal plasticity and imposes further local deformation of the DNA through formation of the stable enzyme-substrate complex. Mechanistically, local distortion augmentation, translation/rotational register shift and partial opening of the nucleosome are employed by AAG to cope with substrate sites in fully exposed, occluded and completely buried positions, respectively. Our findings reveal the molecular basis for the DI-induced modification on the structural dynamics of the nucleosome and elucidate how the DNA glycosylase AAG accesses damaged sites on the nucleosome with different solution accessibility.

Structural remodeling of ribosome associated Hsp40-Hsp70 chaperones during co-translational folding.

Ribosome associated complex (RAC), an obligate heterodimer of HSP40 and HSP70 (Zuo1 and Ssz1 in yeast), is conserved in eukaryotes and functions as co-chaperone for another HSP70 (Ssb1/2 in yeast) to facilitate co-translational folding of nascent polypeptides. Many mechanistic details, such as the coordination of one HSP40 with two HSP70s and the dynamic interplay between RAC-Ssb and growing nascent chains, remain unclear. Here, we report three sets of structures of RAC-containing ribosomal complexes isolated from Saccharomyces cerevisiae. Structural analyses indicate that RAC on the nascent-chain-free ribosome is in an autoinhibited conformation, and in the presence of a nascent chain at the peptide tunnel exit (PTE), RAC undergoes large-scale structural remodeling to make Zuo1 J-Domain more accessible to Ssb. Our data also suggest a role of Zuo1 in orienting Ssb-SBD proximal to the PTE for easy capture of the substrate. Altogether, in accordance with previous data, our work suggests a sequence of structural remodeling events for RAC-Ssb during co-translational folding, triggered by the binding and passage of growing nascent chain from one to another.

Studies of the Sulfonated Hydrogenated Styrene-Isoprene-Styrene Block Copolymer and Its Surface Properties, Cytotoxicity, and Platelet-Contacting Characteristics.

Styrenic thermoplastic elastomers (TPEs) consist of styrenic blocks. They are connected with other soft segments by a covalent linkage and are widely used in human life. However, in biomedical applications, TPEs need to be chemically hydrogenated in advance to enhance their properties such as strong UV/ozone resistance and thermal-oxidative stability. In this study, films composed of sulfonated hydrogenated TPEs were evaluated. Hydrogenated tert-butyl styrene-styrene-isoprene block copolymers were synthesized and selectively sulfonated to different degrees by reaction with acetyl sulfate. By controlling the ratio of the hydrogenated tert-butyl styrene-styrene-isoprene block copolymer and acetyl sulfate, sulfonated films were optimized to demonstrate sufficient mechanical integrity in water as well as good biocompatibility. The thermal plastic sulfonated films were found to be free of cytotoxicity and platelet-compatible and could be potential candidates in biomedical film applications such as wound dressings.

Sulfonation and Characterization of Tert-Butyl Styrene/Styrene/Isoprene Copolymer and Polypropylene Blends for Blood Compatibility Applications.

Hydrogenated styrenic block copolymers (HSBCs) have been used in medical tubing for many years due to their high clarity, flexibility, kink resistance, and toughness. However, when it comes to blood storage applications, HSBC compounds' market has been limited because of their high hydrophobicity, which may trigger platelet adhesion when contacting with blood. HSBC needs to be physically or chemically modified in advance to make it blood compatible; however, HSBC has strong UV/ozone resistance, thermooxidative stability, and excellent processing capability, which increases the difficulty of the chemical modification process as unsaturated dienes has been converted to saturated stable midblocks. Moreover, medical HSBC-containing compounds primarily make up with the non-polar, hydrophobic nature and benign characteristics of other common ingredients (U.S. Pharmacopeia (USP) grades of mineral oil and polypropylene), which complicates the realization of using HSBC-containing compounds in blood-contacting applications, and this explains why few studies had disclosed chemical modification for biocompatibility improvement on HSBC-containing compounds. Sulfonation has been reported as an effective way to improve the material's blood/platelet compatibility. In this study, hydrogenated tert-butyl styrene (tBS)-styrene-isoprene block copolymers were synthesized and its blends with polypropylene and USP grades of mineral oil were selectively sulfonated by reaction with acetyl sulfate. By controlling the ratio of the hydrogenated tBS-styrene-isoprene block copolymer in the blend, sulfonated films were optimized to demonstrate sufficient physical integrity in water as well as thermal stability, hydrophilicity, and platelet compatibility.

Effects of multidisciplinary Internet-based program on management of heart failure.

OBJECTIVES: To develop and test the effectiveness of an Internet-based self management program by multidisciplinary health care professionals for patients with heart failure (HF). METHODS: The comprehensive educational material for HF was created and posted on a website. A computer with Internet connection and computer training were provided first. A secure and simple web-based recording system of vital signs and health behaviors and a mechanism for feedback regarding each participant's record were developed. A randomized controlled trial with a one-year intervention was conducted using a total of 40 patients who were assessed three times in their homes. An intention-to-treat analysis used multivariate statistics. RESULTS: The treatment group had a high (85%) adherence to the intervention. Only the treatment group showed a significant improvement in the knowledge level (p < 0.001), amount of exercise (p = 0.001), and quality of life (p = 0.001), and reduction in HF related symptoms (dyspnea, p = 0.001; fatigue, p = 0.003; functional emotion, p < 0.001), blood pressure (systolic, p = 0.002; diastolic, p < 0.001), frequency of emergency room visit, and length of hospital stay (both p = 0.001). CONCLUSIONS: An effective program to change one's behaviors in managing HF takes a multidisciplinary approach to create and provide feedback regarding a patient's daily record, which can be accomplished through Internet use.