Efficient Homologous Recombination in Mice Using Long Single Stranded DNA and CRISPR Cas9 Nickase.

The CRISPR/Cas9 nickase mutant is less prone to off-target double-strand (ds)DNA breaks than wild-type Cas9 because to produce dsDNA cleavage it requires two guide RNAs to target the nickase to nearby opposing strands. Like wild-type Cas9 lesions, these staggered lesions are repaired by either non-homologous end joining or, if a repair template is provided, by homologous recombination (HR). Here, we report very efficient (up to 100%) recovery of heterozygous insertions in Mus musculus produced by long (>300 nt), single-stranded DNA donor template-guided repair of paired-nickase lesions.

Discovery and identification of fatigue-related biomarkers in human saliva.

OBJECTIVE: To identify stable and specific biomarkers/biomarker combinations for fatigue assessment and establish a discriminant model. PATIENTS AND METHODS: Saliva was collected and electroencephalogram analysis was performed for 47 emergency physicians while awake and after continuoutas duty for 18-24 h. Physicians were divided into the fatigue and non-fatigue groups. Protein spectra of completely quantified saliva specimens were identified before and after long working hours using mass spectrometry. Data were analyzed through Proteome Discoverer software combined with SEQUEST to search protein databases. Proteins were characterized by collision-induced dissociation spectra. A global internal standard (GIS) was added to each group of samples and labeled by tandem mass tags m/z 131.1. All data were compared with GIS, and data between groups were further compared. Qualitative and quantitative data on proteins were exported for fatigue-related proteomic analysis, and a fatigue assessment model was established. RESULTS: We identified 767 salivary proteins in the fatigue group. The correct rates of the discriminant function of the non-fatigue and fatigue groups were 97.1% and 91.7%, respectively (the total correct rate was 95.7%). CONCLUSIONS: We identified 30 fatigue-related protein markers from saliva. We also established a fatigue assessment model for emergency physicians using salivary biomarkers.

Anionic and Cationic Silver Nanoparticle Binding Restructures Net-Anionic PC/PG Monolayers with Saturated or Unsaturated Lipids.

We have examined the interactions between polymer-coated anionic (Ag-COOH) and cationic (Ag-NH) silver nanoparticles, and net-anionic lipid monolayers using dynamic surface pressure measurements. Monolayers composed of saturated or monounsaturated mixtures of anionic phosphatidylglycerol (PG) and zwitterionic phosphatidylcholine (PC) lipids (3:1 molar ratio) were used to determine how lipid packing and monolayer phase state influence the extent of nanoparticle binding and the monolayer response. Anionic Ag-COOH inserted into saturated dipalmitoyl-PC/PG (DPPC/DPPG) and dioleoyl-PC/PG (DOPC/DOPG) monolayers at a low initial surface pressure (10 mN m-1) and caused lipid condensation at high initial surface pressures (20 and 30 mN m-1). Hydrophobic interactions were responsible for insertion, while electrostatic and charge-dipole interactions with PCs were responsible for condensation. In contrast, cationic Ag-NH inserted only into saturated DPPC/DPPG monolayers and otherwise led to lipid condensation. For Ag-NH, adsorption was driven primarily by electrostatic interactions with PGs. Analysis of the subphase Ag and phosphorus concentrations confirmed that Ag-NH had a higher degree binding compared to Ag-COOH, and that the monolayer response was not due to lipid extraction.

Effect of unilateral load carriage on postures and gait symmetry in ground reaction force during walking.

Unilateral load carriage is more hazardous to the musculoskeletal system than bilateral load. The purpose of this study was to examine the effect of such asymmetric carriage on postures and gait symmetry in ground reaction force (GRF) during walking. Kinematics and GRF of 19 adults were recorded while they walked under five load conditions: no load, dumbbell (10 and 20% body weight) held in right and left hand, respectively. After loading, the trunk bent towards the loaded or unloaded side in right- and left-hand trials and under different load weight conditions. The amplitude of trunk bend increased with load, accompanied by decreased stride width, progressively inclined legs towards unloaded side and higher level of asymmetry in medial/lateral GRF (GRFm/l) and free vertical moment GRF (GRFm). The findings indicate the postural adjustment is likely related to the characteristics of load and the task experience and handedness of subject and the unilateral load increases the gait asymmetry in GRFm/l and GRFm.

[Changes of erythrocyte rheology properties in immigrant subjects at different altitudes and its mechanism].

AIM: To explore the influence of altitude changes on red blood cell rheology properties and negative effects caused by it and possible mechanism. METHODS: Erythrocyte filtration index (EFI), superoxide dismutase (SOD) and Malondialdehyde (MDA) were detected the mechanisms in which EFI changes. RESULTS: EFI increased with increasing altitudes and there was a significant different in EFI at the different range of Hb at the same altitude and a significant different in EFI at the similar range of Hb at different altitudes. Correlative analyses showed that EFI was in a negative correlation with SOD activities, relations between EFI and MDA were on the contrary. CONCLUSION: Changes of SOD and MDA with increasing altitudes lead to abnormalities of erythrocyte rheology properties in healthy subjects.