Efficient ssODN-Mediated Targeting by Avoiding Cellular Inhibitory RNAs through Precomplexed CRISPR-Cas9/sgRNA Ribonucleoprotein.

Combined with CRISPR-Cas9 technology and single-stranded oligodeoxynucleotides (ssODNs), specific single-nucleotide alterations can be introduced into a targeted genomic locus in induced pluripotent stem cells (iPSCs); however, ssODN knockin frequency is low compared with deletion induction. Although several Cas9 transduction methods have been reported, the biochemical behavior of CRISPR-Cas9 nuclease in mammalian cells is yet to be explored. Here, we investigated intrinsic cellular factors that affect Cas9 cleavage activity in vitro.

Targeted Disruption of HLA Genes via CRISPR-Cas9 Generates iPSCs with Enhanced Immune Compatibility.

Induced pluripotent stem cells (iPSCs) have strong potential in regenerative medicine applications; however, immune rejection caused by HLA mismatching is a concern. B2M gene knockout and HLA-homozygous iPSC stocks can address this issue, but the former approach may induce NK cell activity and fail to present antigens, and it is challenging to recruit rare donors for the latter method. Here, we show two genome-editing strategies for making immunocompatible donor iPSCs.

Kallikrein 6 secreted by oligodendrocytes regulates the progression of experimental autoimmune encephalomyelitis.

Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS), and experimental autoimmune encephalomyelitis (EAE) is a well-established animal model of the disease. Here, we examined the pathophysiological role of Kallikrein 6 (Klk6), a serine protease produced by oligodendrocytes (OLs), in EAE using Klk6 knockout (Klk6-/-) mice. Compared with Klk6+/+ (wild-type) mice, Klk6-/- mice showed milder EAE symptoms, including delayed onset and milder paralysis.

[Analysis of the Squamous Cell Carcinoma Antigen (SCCA) Which Has a Significant Difference in Values between CLIA and FEIA--Case Report of Extra Glycosylation in SCCA].

Squamous cell carcinoma antigen (SCCA) is a glycoprotein that belongs to the serine protease inhibitor family. Clinically, it has been utilized as a tumor marker for squamous cell carcinoma. In clinical laboratories, SCCA is measured by several immunoassays.

A meta-analytic review of overgeneral memory: The role of trauma history, mood, and the presence of posttraumatic stress disorder.

A number of studies suggest that a history of trauma, depression, and posttraumatic stress disorder (PTSD) are associated with autobiographical memory deficits, notably overgeneral memory (OGM). However, whether there are any group differences in the nature and magnitude of OGM has not been evaluated. Thus, a meta-analysis was conducted to quantify group differences in OGM.

A meta-analysis of cognitive impairment and decline associated with adjuvant chemotherapy in women with breast cancer.

A meta-analysis was performed to quantify the magnitude and nature of the association between adjuvant chemotherapy and performance on a range of cognitive domains among breast cancer patients. A total of 27 studies (14 cross-sectional, 8 both cross-sectional and prospective, and 5 prospective) were included in the analyses, involving 1562 breast cancer patients who had undergone adjuvant chemotherapy and 2799 controls that included breast cancer patients who did not receive adjuvant chemotherapy. A total of 737 effect sizes (Cohen's d) were calculated for cross-sectional and prospective longitudinal studies separately and classified into eight cognitive domains.

The role of childhood and adulthood trauma and appraisal of self-discrepancy in overgeneral memory retrieval.

The aim of this study was to examine the contributory role of childhood and adulthood trauma events and the appraisal of self-discrepancy (the magnitude and distress) in overgeneral memory retrieval (OGM) using a non-clinical sample. Participants with a history of childhood trauma (n=29); adulthood trauma only (n=17) or no-trauma (n=26) participated in this study. The childhood trauma group showed a significantly higher level of general negative memory retrieval than the no-trauma group.

Kinetic study of the daytime atmospheric fate of (Z)-3-hexenal.

Rate coefficients for three daytime atmospheric reactions of (Z)-3-hexenal (3HA)-photolysis (J(1)), reaction with OH radicals (k(2)), and reaction with ozone (k(3))-were measured at 760 Torr and 298 K using a 6 m(3) photochemical reaction chamber. The UV absorption cross sections (σ(3HA)(λ)) were obtained in the wavelength range 240-350 nm. The photodissociation rate of 3HA relative to that of NO(2) was measured by a solar simulator at 760 Torr and was determined to be J(1)/J(NO2) = (4.

A new type of lectin discovered in a fish, flathead (Platycephalus indicus), suggests an alternative functional role for mammalian plasma kallikrein.

A skin mucus lectin exhibiting a homodimeric structure and an S-S bond between subunits of ~40 kDa was purified from flathead Platycephalus indicus (Scorpaeniformes). This lectin, named FHL (FlatHead Lectin), exhibited mannose-specific activity in a Ca(2+)-dependent manner. Although FHL showed no homology to any previously reported lectins, it did exhibit ~20% identity to previously discovered plasma kallikreins and coagulation factor XIs of mammals and Xenopus laevis.

Preliminary investigation of misconceptions and expectations of the effects of traumatic brain injury and symptom reporting.

Objective: This study aimed, first, to introduce a new measure for examining misconceptions of the effects of traumatic brain injury (TBI) and, secondly, to conduct a preliminary investigation of the impact of misconceptions and expectations on level of symptom reporting.

Methods: The Head Injury Knowledge Scale (HIKS) was piloted for its utility in this study using a sample (n = 13) of individuals with brain injury. A sample of 99 uninjured participants was randomly allocated into either TBI simulation (n = 59) or control (n = 40) groups using a 3:2 allocation ratio.

Dark-induced mRNA instability involves RNase E/G-type endoribonuclease cleavage at the AU-box and SD sequences in cyanobacteria.

Light-responsive gene expression is crucial to photosynthesizing organisms. Here, we studied functions of cis-elements (AU-box and SD sequences) and a trans-acting factor (ribonuclease, RNase) in light-responsive expression in cyanobacteria. The results indicated that AU-rich nucleotides with an AU-box, UAAAUAAA, just upstream from an SD confer instability on the mRNA under darkness.

Characterization of two novel mutations of the antithrombin gene observed in Japanese thrombophilic patients.

We investigated the molecular basis of reduced functional levels of antithrombin (AT) in two individuals suffering from thromboembolic events. In each case direct sequencing of amplified DNA revealed 13,260-13,262 del in one patient and 2511C>A in the other patient, predicting a heterozygous E381del and P16H, respectively. Both patients had no 20210A allele and factor V Leiden mutation.

Identification of simultaneous mutation of fibrinogen alpha chain and protein C genes in a Japanese kindred.

Afibrinogenaemia usually induces a bleeding tendency during infancy, whereas protein C deficiency increases susceptibility to thrombosis in children or adolescence. Mutations of these genes have been, therefore, established as independent risk factors for coagulation disorders. We describe the homozygous mutation of the fibrinogen alpha chain gene and additional heterozygous mutation of the protein C gene in a male infant who showed prolonged umbilical bleeding after birth.