Mass Spectrometry Study of Reactive Species in a Microhollow Cathode Discharge in He+H2O Mixtures

Reactive species created by a microhollow cathode discharge (MHCD) in He+H2O mixtures were investigated at 160 Torr using the molecular beam mass spectrometry. Ion currents of He+, HeH+, O+, OH+, H+(H2O), and H+(H2O)2 were measured as functions of H2O relative concentration (0.24 - 14%) and discharge current (5 - 15 mA). When the concentration exceeds 6%, most of the ions were decreased, but H+(H2O)2 (cluster ion) significantly increased at discharge current of 5 mA, whose ion current was very low compared with those of other ions. With increasing the discharge current, the cluster ion showed a sharp decrease, while the other ions were almost constant. These features were reasonably explained by the increase of the gas temperature and the plasma rarefaction due to the Joule heating of the working gas

Therapeutic potential of PRL-3 targeting and clinical significance of PRL-3 genomic amplification in gastric cancer

<p>Abstract</p> <p>Background</p> <p>Phosphatase of regenerating liver-3 (PRL-3) has deserved attention as a crucial molecule in the multiple steps of metastasis. In the present study, we examined the mechanisms regulating PRL-3 expression, and assessed the clinical potential of PRL-3-targeted therapy in gastric cancer.</p> <p>Methods</p> <p>PRL-3 genomic amplification was analyzed using quantitative-polymerase chain reaction and/or fluorescence in situ hybridization in 77 primary gastric tumors. The anticancer activity of PRL-3 inhibitor (1-4-bromo-2-benzylidene rhodanine) treatment was evaluated against cancer cells with different genetic and expression status.</p> <p>Results</p> <p>PRL-3 genomic amplification was closely concordant with high level of its protein expression in cell lines, and was found in 20% (8/40) among human primary tumors with its expression, which were all stage III/IV disease (40%, 8/20), but in none (0/37) among those without expression. Additionally, PRL-3 genomic amplification was associated with metastatic lymph node status, leading to advanced stage and thereby poor outcomes in patients with lymph node metastasis (<it>P </it>= 0.021). PRL-3 small interfering RNA robustly repressed metastatic properties, including cell proliferation, invasion, and anchorage-independent colony formation. Although neither PRL-3 genomic amplification nor expression level was responsible for the sensitivity to PRL-3 inhibitor treatment, the inhibitor showed dose-dependent anticancer efficacy, and remarkably induced apoptosis on all the tested cell lines with PRL-3 expression.</p> <p>Conclusions</p> <p>We have for the first time, demonstrated that PRL-3 genomic amplification is one of the predominant mechanisms inducing its expression, especially in more advanced stage, and that PRL-3-targeted therapy may have a great potential against gastric cancer with its expression.</p

Population pharmacokinetic modeling of GS‐441524, the active metabolite of remdesivir, in Japanese COVID‐19 patients with renal dysfunction

腎障害患者におけるレムデシビルの薬物動態モデルを構築 --新型コロナウイルス感染症治療薬の適正使用に向けて--. 京都大学プレスリリース. 2021-11-25.Remdesivir, a prodrug of the nucleoside analog GS-441524, plays a key role in the treatment of coronavirus disease 2019 (COVID-19). However, owing to limited information on clinical trials and inexperienced clinical use, there is a lack of pharmacokinetic (PK) data in patients with COVID-19 with special characteristics. In this study, we aimed to measure serum GS-441524 concentrations and develop a population PK (PopPK) model. Remdesivir was administered at a 200 mg loading dose on the first day followed by 100 mg from day 2, based on the package insert, in patients with an estimated glomerular filtration rate (eGFR) greater than or equal to 30 ml/min. In total, 190 concentrations from 37 Japanese patients were used in the analysis. The GS-441524 trough concentrations were significantly higher in the eGFR less than 60 ml/min group than in the eGFR greater than or equal to 60 ml/min group. Extracorporeal membrane oxygenation in four patients hardly affected the total body clearance (CL) and volume of distribution (Vd) of GS-441524. A one-compartment model described serum GS-441524 concentration data. The CL and Vd of GS-441524 were significantly affected by eGFR readjusted by individual body surface area and age, respectively. Simulations proposed a dose regimen of 200 mg on day 1 followed by 100 mg once every 2 days from day 2 in patients with an eGFR of 30 ml/min or less. In conclusion, we successfully established a PopPK model of GS-441524 using retrospectively obtained serum GS-441524 concentrations in Japanese patients with COVID-19, which would be helpful for optimal individualized therapy of remdesivir

Pharmacist-physician collaborative care for outpatients with left ventricular assist devices using a cloud-based home medical management information-sharing system: a case report

[Background] The standard anticoagulation therapy for patients implanted with left ventricular assist devices (LVADs) includes warfarin therapy. We developed a cloud-based home medical management information-sharing system named as LVAD@home. The LVAD@home system is an application designed to be used on iPad tablet computers. This system enables the sharing of daily information between a patient and care providers in real time. In this study, we reported cases of outpatients with LVADs using this system to manage anticoagulation therapy. [Case presentation] The patient, a man in his 40s with end-stage heart failure owing to non-ischemic dilated cardiomyopathy, underwent LVAD implantation and warfarin was started on postoperative day 1. He started to use LVAD@home to manage warfarin therapy after discharge (postoperative day 47). He sent his data to care providers daily. By using this system, the pharmacist observed his signs of reduced dietary intake 179 days after discharge, and after consulting the physician, told the patient to change the timing of the next measurement earlier than usual. On the next day, the prothrombin time-international normalized ratio increased from 2.0 to 3.0, and thus the dose was decreased by 0.5 mg. Four patients used this system to monitor warfarin therapy from October 2015 to March 2018. In these patients, the time in therapeutic range was 90.1 ± 1.3, which was higher than that observed in previous studies. Additionally, there were no thromboembolic events or bleeding events. [Conclusions] The cloud-based home management system can be applied to share real-time patient information of factors, including dietary intake that interact with warfarin. It can help to improve long-term anticoagulation outcomes in patients implanted with LVAD

合併症を有するB型大動脈解離に対するステントグラフト内挿術における腎動脈に対する治療戦略 : 多施設共同研究

Background: Management of abdominal branches associated with Stanford type B aortic dissection is controversial without definite criteria for therapy after thoracic endovascular aortic repair (TEVAR). This is in part due to lack of data on natural history related to branch vessels and their relationship with the dissection flap, true lumen, and false lumen. Purpose: To investigate the natural history of abdominal branches after TEVAR for type B aortic dissection and the relationship between renal artery anatomy and renal volume as a surrogate measure of perfusion. Materials and Methods: This study included patients who underwent TEVAR for complicated type B dissection from January 2012 to March 2017 at 20 centers. Abdominal aortic branches were classified with following features: patency, branch vessel origin, and presence of extension of the aortic dissection into a branch (pattern 1, supplied by the true lumen without branch dissection; pattern 2, supplied by the true lumen with branch dissection, etc). The branch artery patterns before TEVAR were compared with those of the last follow-up CT (mean interval, 19.7 months) for spontaneous healing. Patients with one kidney supplied by pattern 1 and the other kidney by a different pattern were identified, and kidney volumes over the course were compared by using a simple linear regression model. Results: Two hundred nine patients (mean age ± standard deviation, 66 years ± 13; 165 men and 44 women; median follow-up, 18 months) were included. Four hundred fifty-nine abdominal branches at the last follow-up were evaluable. Spontaneous healing of the dissected branch occurred in 63% (64 of 102) of pattern 2 branches. Regarding the other patterns, 6.5% (six of 93) of branches achieved spontaneous healing. In 79 patients, renal volumes decreased in kidneys with pattern 2 branches with more than 50% stenosis and branches supplied by the aortic false lumen (patterns 3 and 4) compared with contralateral kidneys supplied by pattern 1 (pattern 2 vs pattern 1: −16% ± 16 vs 0.10% ± 11, P = .002; patterns 3 and 4 vs pattern 1: −13% ± 14 vs 8.5% ± 14, P = .004). Conclusion: Spontaneous healing occurs more frequently in dissected branches arising from the true lumen than in other branch patterns. Renal artery branches supplied by the aortic false lumen or a persistently dissected artery with greater than 50% stenosis are associated with significantly greater kidney volume loss.博士（医学）・乙第1461号・令和2年6月30日Copyright © 2019 by authors and RSNA. This work is licensed under the Creative Commons Attribution International License (CC BY-NC-ND 4.0). https://creativecommons.org/licenses/by-nc-nd/4.0/

RINL, Guanine Nucleotide Exchange Factor Rab5-Subfamily, Is Involved in the EphA8-Degradation Pathway with Odin

The Rab family of small guanosine triphosphatases (GTPases) plays a vital role in membrane trafficking. Its active GTP-bound state is driven by guanine nucleotide-exchange factors (GEFs). Ras and Rab interactor (or Ras interaction/interference)-like (RINL), which contains a conserved VPS9 domain critical for GEF action, was recently identified as a new Rab5 subfamily GEF in vitro. However, its detailed function and interacting molecules have not yet been fully elucidated. Here we found that RINL has GEF activity for the Rab5 subfamily proteins by measuring their GTP-bound forms in cultured cells. We also found that RINL interacts with odin, a member of the ankyrin-repeat and sterile-alpha motif (SAM) domain-containing (Anks) protein family. In addition, the Eph tyrosine kinase receptor EphA8 formed a ternary complex with both RINL and odin. Interestingly, RINL expression in cultured cells reduced EphA8 levels in a manner dependent on both its GEF activity and interaction with odin. In addition, knockdown of RINL increased EphA8 level in HeLa cells. Our findings suggest that RINL, as a GEF for Rab5 subfamily, is implicated in the EphA8-degradation pathway via its interaction with odin

Structure of RINL.

<p>(A) Diagram of the structural features of RIN family members. The lower numbers represent the amino acid residues. (B) FLAG-RIN1, RIN2, RIN3, and RINL were transiently co-transfected with myc-amphiphysin II (amph II) into HEK293T cells. Cells lysates were immunoprecipitated with anti-FLAG antibody, followed by immunoblotting with anti-myc and anti-FLAG antibodies. Total lysates were immunoblotted with anti-myc antibody. (C) Cell lysates from HEK293T cells were applied to a Superdex 200 Prep Grade gel filtration column. The elution position was compared with those of the globular size markers (upper panel). The fractions (0.5 ml) eluted from the column and total lysate (tot.) were analyzed by SDS-PAGE, and proteins were immunoblotted with anti-RINL antibody.</p

GEF activity of RINL for Rab5 subfamily proteins.

<p>(A–D) HEK293T cells expressing myc-Rab5b (A), Rab21 (B), Rab22 (C), or Rab31 (D) and FLAG-mock, RINL, RIN3, or Rabex-5 were metabolically radiolabeled with ³²P_i for 4 hours. Myc-Rab5 subfamily proteins were immunoprecipitated with an anti-myc monoclonal antibody, and nucleotides associating with each Rab protein were separated by thin-layer chromatography. The radioactivity of GTP and GDP was quantified, and the percentages (%) of each GTP-bound Rab are shown. Total lysates (bottom) and immunoprecipitated samples (middle) from the radiolabeled cells were separated by SDS-PAGE and immunoblotted with anti-FLAG and anti-myc antibodies, respectively. *p<0.05 vs. mock-transfected cells. (E) Myc-Rab3a, 7a, or 11a was co-transfected with FLAG-mock or RINL into HEK293T cells. The percentages of each GTP-bound Rab member in the metabolically radiolabeled cells are shown as described in (A). (F) Myc-Rab5b was co-transfected with wild type (WT), or the DP_AA or YT_AA mutant of FLAG-RINL into HEK293T cells. The percentages of GTP-Rab5b in the metabolically radiolabeled cells are shown as described in (A). Total lysates (bottom) and immunoprecipitated samples (middle) from the radiolabeled cells were separated by SDS-PAGE and immunoblotted with anti-FLAG and anti-myc antibodies, respectively. All data were obtained from more than three independent experiments and are shown as the mean ± S.E. (error bars). **p<0.01 vs. mock-transfected cells.</p

Identification of odin/Anks1a as an interacting molecule with RINL.

<p>(A) HeLa cell lysates were immunoprecipitated with normal rat IgG or anti-odin antibody, followed by immunoblotting with antibodies as indicated. (B) FLAG-RIN family or FLAG-mock were transfected into HEK293T cells. Cells lysates were immunoprecipitated with anti-FLAG antibody, followed by immunoblotting with antibodies as indicated. (C) FLAG-RINL and the indicated deletion mutants of myc-odin were transiently transfected into HEK293T cells. Cells lysates were immunoprecipitated with anti-myc antibody, followed by immunoblotting with antibodies as shown. (D) The indicated deletion mutants of FLAG-RINL were transiently transfected into HEK293T cells. Cells lysates were immunoprecipitated with anti-FLAG antibody, followed by immunoblotting with antibodies as indicated. (E) Myc-odin and V5-RINL were co-transfected with FLAG-tagged constitutively active (CA, lanes 2 and 4) or mock (lanes 1 and 3) into HEK293T cells. Cell lysates were immunoprecipitated with anti-myc antibody, followed by immunoblotting with antibodies as indicated. Aliquots of total lysates were also immunoblotted with antibodies as indicated.</p