Genome editing in mammalian cells using the CRISPR type I-D nuclease

Adoption of CRISPR–Cas systems, such as CRISPR–Cas9 and CRISPR–Cas12a, has revolutionized genome engineering in recent years; however, application of genome editing with CRISPR type I—the most abundant CRISPR system in bacteria—remains less developed. Type I systems, such as type I-E, and I-F, comprise the CRISPR-associated complex for antiviral defense (‘Cascade’: Cas5, Cas6, Cas7, Cas8 and the small subunit) and Cas3, which degrades the target DNA; in contrast, for the sub-type CRISPR–Cas type I-D, which lacks a typical Cas3 nuclease in its CRISPR locus, the mechanism of target DNA degradation remains unknown. Here, we found that Cas10d is a functional nuclease in the type I-D system, performing the role played by Cas3 in other CRISPR–Cas type I systems. The type I-D system can be used for targeted mutagenesis of genomic DNA in human cells, directing both bi-directional long-range deletions and short insertions/deletions. Our findings suggest the CRISPR–Cas type I-D system as a unique effector pathway in CRISPR that can be repurposed for genome engineering in eukaryotic cells

Medium-firm drug-candidate library of cryptand-like structures on T7: Design and selection of strong binder for Hsp90

We designed and synthesized a medium-firm drug-candidate library of cryptand-like structures possessing a randomized peptide linker on the bacteriophage T7. From the macrocyclic library with a 10^9 diversity, we obtained a binder toward a cancer-related protein (Hsp90) with an antibody-like strong affinity (K_D = 62 nM) and the binding was driven by the enthalpy. The selected supramolecular ligand inhibited Hsp90 activity by site-specific binding outside of the well-known ATP-binding pocket on the N-terminal domain (NTD)

Usefulness of Serum C-reactive Protein in the Management of Adult Community-acquired Pneumonia

C-Reactive protein (CRP) is widely used as a marker of infection, but there is insufficient evidence as to its usefulness in patients with community-acquired pneumonia (CAP). In the present study, we investigated the clinical usefulness of CRP in a retrospective study of 242 patients aged 14 years who were hospitalized with CAP. Patients were classified into three groups according to the number of days between disease onset and the initial measurement of CRP as follows: Group 1, 0-1 day; Group 2, 2-4 days; Group 3, 5 days. Patients in Groups 2 and 3, who had more severe pneumonia, had higher CRP levels. Over time, CRP levels decreased in the responders in Groups 2 and 3; specifically, in Group 2, median CRP levels on Days 0, 3, and 7 were 9.85, 5.33, and 0.81mg/dL, respectively, compared with 9.99, 4.29, and 0.70mg/dL, respectively, in Group 3. In patients not responding to initial treatment, median CRP levels increased from Day 0 to Day 3 (4.32 vs. 11.70mg/dL, respectively). In all non-responders, CRP levels on Day 3 were>50% of levels on Day 0. In conclusion, when measured approximately 48 h after disease onset, CRP is useful for evaluating the severity of pneumonia and predicting the response to treatment. A good clinical outcome is likely when CRP levels on Day 3 are 50% of those on admission

Percutaneous Transcatheter Ballon Valvuloplasty of a Stenosed Bioprosthetic Tricuspid Valve

Article信州医学雑誌 67(5): 289-292(2019)journal articl

Q fever in acute upper respiratory tract infection

We examined whether or not acute upper respiratory tract infection is associated with Q fever (Coxiella burnetii infection). The subjects consisted of 124 patients with acute upper respiratory tract infection. At initial medical consultation, the presence or absence of serum C. burnetii was examined by nested PCR method. Of the 124 patients, no patients (0 percent) were positive for C. burnetii in serum. These results suggested that the involvement of Q fever in acute upper respiratory tract infection is extremely low

Clinical effect of 3g/day administration of meropenem on severe pneumonia

We examined the clinical effect of Meropenem (MEPM) on severe pneumonia. We administered 3g of Meropenem daily to 20 patients with severe pneumonia: 8 communityacquired pneumonia patients, 9 nursing and healthcare-associated pneumonia patients, and 3 hospital-acquired pneumonia patients. It was effective in 15 of the 20 patients (75%): 8 of 8 community-acquired pneumonia patients (100%), 6 of 9 nursing and healthcare-associated pneumonia patients (66.6%), and 1 of 3 hospital-acquired pneumonia patients (33.3%). Bacteriologically, 9 of a total of 10 strains (90%) were eradicated: 4 of 4 Streptococcus pneumoniae strains, 2 of 2 methicillin-sensitive Staphlococcus aureus strains, 1 of 2 Enterococcus faecalis strains, 1 of 1 Klebsiella pneumoniae strain, and 1 of 1 Escherichia coli strain. Hepatic dysfunction was observed as a side effect in 8 patients (40%). Based on the above, administration of MEPM daily 3 g is extremely effective for community-acquired pneumonia, while it appears ineffective in many cases of nursing and healthcare-associated pneumonia or hospital-acquired pneumonia, and results in hepatic dysfunction at a high frequency