CRISPR RNAs trigger innate immune responses in human cells

Here, we report that CRISPR guide RNAs (gRNAs) with a 5'-triphosphate group (5'-ppp gRNAs) produced via in vitro transcription trigger RNA-sensing innate immune responses in human and murine cells, leading to cytotoxicity. 5'-ppp gRNAs in the cytosol are recognized by DDX58, which in turn activates type I interferon responses, causing up to similar to 80% cell death. We show that the triphosphate group can be removed by a phosphatase in vitro and that the resulting St-hydroxyl gRNAs in complex with Cas9 or Cpfl avoid innate immune responses and can achieve targeted mutagenesis at a frequency of 95% in primary human CD4(+) T cells. These results are in line with previous findings that chemically synthesized sgRNAs with a 5'-hydroxyl group are much more efficient than in vitro-transcribed (IVT) sgRNAs in human and other mammalian cells. The phosphatase treatment of IVT sgRNAs is a cost-effective method for making highly active sgRNAs, avoiding innate immune responses in human cells.

Distinct vertical behavior of key Arctic copepods following the midnight sun period in the East Siberian continental margin region, Arctic Ocean

Diel vertical migration (DVM) of zooplankton plays a vital role in biological carbon pump and food web interactions. However, there is considerable debate about the DVM of zooplankton in response to environmental changes in the Arctic Ocean. We investigated DVM behavior in the key Arctic copepods Calanus glacialis, Calanus hyperboreus, and Metridia longa following the midnight sun period in the East Siberian continental margin region. The two Calanus species showed non-DVM behaviors, whereas M. longa showed a typical DVM pattern consistent with the solar radiation cycle. Additionally, these species showed different vertical distributions. Calanus glacialis was distributed at depths above 20 m in the warm fresh water, where the highest density gradient was observed. Calanus hyperboreus was distributed at depths between 30 and 55 m in the cold salty water, where a high contribution of micro phytoplankton and the subsurface chlorophyll maximum (SCM) layer were observed. M. longa was found across a broader range of temperature and salinity than both Calanus species, and it was distributed in the upper water column, where the SCM layer was observed at night and at depths between 100 and 135 m in the daytime. These results imply that M. longa can be well adapted to the changing Arctic Ocean environment, where sea ice loss and ocean warming are ongoing, whereas C. hyperboreus can be the most vulnerable to these changes. These findings provide important information for understanding variations in the vertical distributions of key copepod species in the rapidly changing Arctic marine environment

Two cases of pyogenic liver abscess due to Klebsiella pneumoniae in immunocompetent children

Pyogenic liver abscess (PLA) can be caused by bacteria entering the liver via the portal vein or primary bacteremia, or it can be cryptogenic. Recently, Klebsiella pneumoniae has been increasingly found as a PLA pathogen. PLA due to this bacterium often leads to formation of extrahepatic abscesses. The treatment of choice is dual therapy with insertion of percutaneous catheter drainage and antibiotic therapy. We report 2 cases of PLA due to K. pneumoniae in immunocompetent children. We successfully treated patient 1 with percutaneous catheter drainage for 18 days and 6-week course of antibiotic therapy. Patient 2 was treated with percutaneous needle aspiration and antibiotic therapy for the same period. In both patients, the PLAs showed the ultrasound-confirmed resolutions after the dual therapy

Comparative study of the treatment outcomes of osteoporotic compression fractures without neurologic injury using a rigid brace, a soft brace, and no brace : a prospective randomized controlled non-inferiority trial

BACKGROUND:The efficacy of brace application for the treatment of osteoporotic compression fractures remains unclear. The purpose of this study was to compare the treatment outcomes in patients with osteoporotic compression fractures with regard to whether the patients had no braces, rigid braces, or soft braces.METHODS:We randomly assigned sixty patients with acute one-level osteoporotic compression fractures within three days of injury to the no-brace, soft-brace, and rigid-brace groups through 1:1:1 allocation. The primary outcome was the baseline adjusted Oswestry Disability Index score at twelve weeks after compression fracture. The non-inferior margin of the Oswestry Disability Index was set at an average of 10 points.RESULTS:The baseline adjusted Oswestry Disability Index score at twelve weeks after compression fracture in the no-brace group was not inferior to that in the soft-brace or rigid-brace groups. The mean adjusted Oswestry Disability Index score was 35.95 points (95% confidence interval, 25.42 to 46.47 points) in the no-brace group and 37.83 points (95% confidence interval, 26.77 to 48.90 points) in the soft-brace group, with a difference of -1.88 points (95% confidence interval, -7.02 to 9.38 points) between the groups. Similarly, the mean adjusted Oswestry Disability Index score was 35.95 points (95% confidence interval, 25.42 to 46.47 points) in the no-brace group and 33.54 points (95% confidence interval, 23.79 to 43.29 points) in the rigid-brace group, with a difference of 2.41 points (95% confidence interval, -7.86 to 9.27 points) between the groups. During the follow-up assessment period, there was no significant difference among the groups for the overall Oswestry Disability Index scores (p = 0.260), visual analog scale for pain scores for back pain (p = 0.292), and anterior body compression ratios (p = 0.237). However, the Oswestry Disability Index scores and the visual analog scale scores for back pain significantly improved with time after the fractures (p < 0.001), and the body compression ratios significantly decreased with time in all three groups (p < 0.001).CONCLUSIONS:The Oswestry Disability Index scores for the treatment of compression fractures without a brace were not inferior to those with soft or rigid braces. Moreover, the improvement in back pain and progression of anterior body compression were similar among the three groups.OAIID:oai:osos.snu.ac.kr:snu2014-01/102/0000004226/15SEQ:15PERF_CD:SNU2014-01EVAL_ITEM_CD:102USER_ID:0000004226ADJUST_YN:NEMP_ID:A079510DEPT_CD:801CITE_RATE:4.309DEPT_NM:의학과SCOPUS_YN:NCONFIRM:

Radioprotective effects of fucoidan on bone marrow cells: improvement of the cell survival and immunoreactivity

Fucoidan is a sulfated polysaccharide purified from brown algae including Fucus vesiculosus and has a variety of biological effects including mobilization of hematopoietic progenitor cells. Recently, we demonstrated that fucoidan stimulates the antigen-presenting functions of dendritic cells. In this study, we investigated the radioprotective effects of fucoidan on bone marrow cells (BMCs), which are the main cellular reservoir for the hematopoietic and immune system. To evaluate the effects of fucoidan, we assayed cell viability and immune responses. In a viability assay, fucoidan significantly increased the viability of BMCs. Based on the results of flow cytometric analysis, the increased viability of fucoidan-treated BMCs was attributed to the inhibition of radiation-induced apoptosis. Furthermore, fucoidan altered the production of immune-related cytokines from BMCs and increased the capability of BMCs to induce proliferation of allogeneic splenocytes. Taken together, our study demonstrated that fucoidan has radioprotective effects on BMCs with respect to cell viability and immunoreactivity. These results may provide valuable information, useful in the field of radiotherapy

Anaplastic Thyroid Carcinoma Initially Presented with Abdominal Cutaneous Mass and Hyperthyroidism

A 63-year-old female was admitted to our hospital with a tender abdominal wall mass about 15 cm in diameter, which she had for 1 month. About 1 week earlier, the patient had also perceived a mass in the neck area. Computed tomography revealed huge thyroid and periumbilical masses. The thyroid hormone levels were consistent with a hyperthyroid state. Pathological examination of the thyroid mass was compatible with anaplastic thyroid carcinoma (ATC) and the abdominal cutaneous mass was shown to be metastatic ATC. Despite palliative radiotherapy and chemotherapy, the patient died of respiratory failure on her 63rd day of hospitalization. This case demonstrates that abdominal cutaneous metastasis and hyperthyroidism can occur as initial manifestations of ATC. To our knowledge, this is the first reported case

Severe Pulmonary Adverse Effects in Lymphoma Patients Treated with Cyclophosphamide, Doxorubicin, Vincristine, and Prednisone (CHOP) Regimen Plus Rituximab

BACKGROUND/AIMS: The aim of our study was to determine the incidence and clinical features of severe pulmonary complications in patients receiving cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) or rituximab plus CHOP (R-CHOP) as the initial treatment for lymphoma. METHODS: A retrospective analysis of pulmonary infection and drug-induced interstitial pneumonitis (DIIP) was performed using lymphoma registry data. R-CHOP was administered in 71 patients and CHOP in 29 patients. RESULTS: The severe pulmonary adverse events tended to occur more frequently with R-CHOP (18.3%) than CHOP alone (13.8%), although the difference was not significant (p = 0.771). DIIP occurred in five patients in the R-CHOP arm (7%) and in one in the CHOP arm (3%). The continuous use of steroids for conditions other than lymphoma significantly increased the risk of pulmonary infection including Pneumocystis jiroveci pneumonia (p = 0.036) in the multivariate analysis. International prognostic index, tumor stage, smoking, previous tuberculosis, chronic obstructive pulmonary disease, and lymphoma involvement of lung parenchyma were not related to pulmonary adverse events. Patients who experienced severe pulmonary events showed shorter survival when compared to those without complications (p = 0.002). CONCLUSIONS: Our experiences with serial cases with DIIP during chemotherapy and the correlation of continuous steroid use with pulmonary infection suggest that the incidence of pulmonary complications might be high during lymphoma treatment, and careful monitoring should be performed.ope