Biochemical Activity of PIF1 Helicase from Thermophilic Bacteria / 中国生物化学与分子生物学报

Petite integration frequency 1 (PIF1) helicases are ubiquitous enzymes which play vital roles in nearly all DNA metabolic processes. In recent years, the biochemical activity and three-dimensional structure of several PIF1 helicases have been reported, but there are few reports on the PIF1 helicase of bacteria living in extreme environments. In this paper, a series of biochemical and biophysical techniques were used to study the Thermodesulfovibrio yellowstonii PIF1 (Ty.PIF1) helicase in many aspects. Ty. PIF1 was obtained with a purity of over 90% and good uniformity using the prokaryotic expression and purification system. Ty.PIF1 is a monomer with a calculated molecular weight of 60 kD in solution. Ty. PIF1 has high thermal stability. The secondary structure remains stable when the temperature is below 65 ℃, and the secondary structure changes only when the temperature is above 70 ℃. The optimal unwinding temperature of Ty.PIF1 in vitro is 45 ℃, which is not the optimal temperature for the survival of thermodesulfovibrio yellowstonii. It indicates that when Ty.PIF1 exerts its enzymatic activity in vivo, it may require the participation of other cofactors. Ty.PIF1 can exert unwinding activity in a wide temperature range (20-55 ℃), and the presence of enzyme activity at 55 ℃ indicates that Ty.PIF1 has heat-resistant properties. Ty.PIF1 prefers to bind to substrates containing ssDNA, but there is certain requirement for the length of the ssDNA, which is at least 4 nt in length. Ty.PIF1 can also bind to the G

Effect of adaptive movement on durability and working time of twisted file

PURPOSE: Recently TF-adaptive movement is developed in order to increase the durability of TF files. The purpose of this study was to assess the effects of adaptive movement on durability and performance of twisted files. MATERIALS AND METHODS: Resin blocks simulating artificial J-shape canals were used for this study. In TFC group, TF-adaptive ML-1 (25/.08 size) files were used to prepare the canals under continuous rotation 500 rpm/4.0 Ncm. In TFA group, TF-adaptive ML-1 (25/.08 size) files were used to prepare the canals under adaptive movement. After preparing each artificial canal, TF files were observed under dental microscope for assessing existence of unwinding, distortion, and fracture. If unwinding of flute was observed, the number of artificial canals until unwinding of flute occurs was recorded. Required time until instruments reach working length and distance of unwinded portion of files from D0 were measured. All test results were conducted by Mann-Whitney U test at a 0.05 level of significance. RESULTS: No NiTi instrument's separation was observed. Number of resin blocks until file unwinding happens and working time was significantly high in TFA group compared to TF group. Distance of distortion from D0 didn't show significant difference between TFA, TF groups. CONCLUSION: The number of resin blocks prepared until unwinding happens and working time were significantly high in TFA group. The location of unwinding showed no significant difference between 2 groups. Adaptive movement increased the number of canals prepared until unwinding occurs and working time of twisted files.

Effects of bisbenzylisoquinoline alkaloid tetrandrine derivative HL-27 on biological properties of BLM helicase / 中国药理学通报

Aim To investigate the effects of bisbenzyl-isoquinoline alkaloid tetrandrine derivative HL-27 on the biological properties of the BLM642-1290 helicase. Methods Fluorescence polarization technique was used to investigate the effects of bisbenzylisoquinoline alkaloid tetrandrine derivative HL-27 on the DNA bind-ing activity and unwinding activity of the BLM642-1290 helicase. Malachite green-phosphate ammonium molyb-date colorimetry was used to investigate the effects of HL-27 on the ATPase activity of the BLM642-1290 heli-case. Ultraviolet spectral scanning was used to investi-gate the effects of HL-27 on the conformation of the BLM642-1290 helicase. Results When the concentra-tion of HL-27 reached 33.34 μmol·L-1, the inhibi-tion ratio of dsDNA and ssDNA binding activity of the BLM642-1290 helicase was 41.35% and 59.54% , re-spectively. When the concentration of HL-27 reached 50 μmol·L-1, the inhibition ratio of DNA unwinding activity of the BLM642-1290 helicase was 78.68% . When the concentration of HL-27 reached 100 μmol· L-1, the inhibition ratio of ATPase activity of the BLM642-1290 helicase was 43.8% . Conclusion The DNA binding activity, ATPase activity and unwinding activity of the BLM642-1290 helicase can be inhibited by bisbenzylisoquinoline alkaloid tetrandrine derivative HL-27.

Lifetime and fracture patterns of NiTi rotary files in molars / 대한치과보존학회지

Intracanal separation of the rotary files is a serious concern in modern endodontic practice. The objective of this study was to compare the life span and fracture patterns of three NiTi rotary files in molar teeth. Mesiobuccal roots of upper molar (n = 150) and mesial roots of lower molar (n = 150) were divided into three groups and each group was prepared with Profile, ProTaper, and K3 respectively. Every file was used until separation and/or deterioration of the cutting blade was happened, and then the number of canals to separation and/or unwinding were recorded. Radiographs and Scanning electon microscope (SEM) photographs were taken to evaluate the patterns of separation. The results were as follows : 1. There were no significant differences in numbers of canals to separation and/or unwinding among the groups. 2. Comparing between flaring files, K3 showed significant lower numbers of canals to separation and/or unwinding (p < 0.05), and there was no significant difference between shaping files 3. Separations of instruments were occurred at the midpoint of curvatures within the canals. 4. In SEM observations, ductile fractures were seen in most of cases, characterized by shallow dimples. Additional researches is needed to provide a new guideline that informs the appropriate number of times to use NiTi files.

The conformations and energetics of photodamaged DNA.

Energy minimization techniques are used in conjunction with the results of small molecule crystallographic studies on relevant compounds to propose structural models for photodamaged DNAs. Specifically, we present models both for a DNA molecule containing a psoralen photo-crosslink and for a DNA molecule containing a thymine photodimer. In both models, significant distortions of the nucleic acid helix are observed, including kinking and unwinding at the damage site and numerous changes in the backbone torsion angles relative to their standard conformations. Both the torsion angle geometries and the energetics of the models are presented in detail.