Добірка наукової літератури з теми "WN 250.6"

ABSTRACT The nucleoside triphosphatase (NTPase)/helicase associated with nonstructural protein 3 of West Nile (WN) virus was purified from cell culture medium harvested from virus-infected Vero cells. The purification procedure included sequential chromatography on Superdex-200 and Reactive Red 120 columns, followed by a concentration step on an Ultrogel hydroxyapatite column. The nature of the purified protein was confirmed by immunoblot analysis using a WN virus-positive antiserum, determination of its NH2 terminus by microsequencing, and a binding assay with 5′-[14C]fluorosulfonylbenzoyladenosine. Under optimized reaction conditions the enzyme catalyzed the hydrolysis of ATP and the unwinding of the DNA duplex with k cat values of 133 and 5.5 × 10−3 s−1, respectively. Characterization of the NTPase activity of the WN virus enzyme revealed that optimum conditions with respect to the Mg2+requirement and the monovalent salt or polynucleotide response differed from those of other flavivirus NTPases. Initial kinetic studies demonstrated that the inhibition (or activation) of ATPase activity by ribavirin-5′-triphosphate is not directly related to changes in the helicase activity of the enzyme. Further analysis using guanine andO 6-benzoylguanine derivatives revealed that the ATPase activity of WN virus NTPase/helicase may be modulated, i.e., increased or reduced, with no effect on the helicase activity of the enzyme. On the other hand the helicase activity could be modulated without changing the ATPase activity. Our observations show that the number of ATP hydrolysis events per unwinding cycle is not a constant value.

Purpose The purpose of this study is to conduct gas tungsten arc dissimilar welding of AISI 304 stainless steel and low carbon steel within a process window so as to investigate the effects of current, speed and gas flow rate (GFR) on the microstructure and mechanical properties of the weldments. Design/methodology/approach The welding experiment was carried out at different combinations of parameters using WN-250S Kaierda electric welding machine. A combination of scanning electron microscopy and energy dispersive X-ray spectroscopy was used to examine the microstructure of the weldments. Micro-hardness and tensile tests were performed using Vickers hardness tester and Instron universal testing machine, respectively. ANOVA was used to analyze the significance of the parameters on the mechanical properties. Findings The microstructure of the weld region is characterized with dendritic structure with the existence of ferrite and austenite phases. The utilized parameters show significant effects on the ultimate tensile strength (UTS) of the weldments. The current and GFR were found to be the most and least significant factors, respectively. Both the grain size and weld penetration contributed to the UTS of the weldments. The UTS (427-886 MPa) increased with decreasing current and welding speed. In all samples, the weld region exhibited higher hardness (297-396 HV) than the HAZ in the base metals (maximum of 223 Â ± 6 HV). All the three factors show significant effect with the welding speed contributing mostly to the hardness of the weld region. Originality/value The parametric combination that gives the optimum mechanical performance of the dissimilar gas tungsten arc weldments of AISI 304 stainless steel and low carbon steel was established.

Background:Patients with systemic lupus erythematosus (SLE) who achieved the clinical state as serologically active clinically quiescent (SACQ). It appears to account for 6–12% of all patients with SLE, but there is disagreement about whether such patients are indeed clinically stable [1-3], especially in Chinese patients. And there is no conclusion as to what kind of treatment should be taken for such patients.Objectives:To clarify the frequency and outcome of SACQ patients in lupus. And to identify factors associated with the flare of disease.Methods:Clinical data of patients diagnosed as SLE and followed in Peking University First Hospital from 2009 to 2015 were retrospectively reviewed. 682 patients with systemic lupus erythematosus who were followed up for more than 6 months at Peking University First Hospital from January 2007 to December 2015 were summarized. SACQ was defined as an at least a 6-month period with persistent serologic activity and without clinical activity and could be taking a daily dose of prednisone or equivalent less than 7.5 mg. Serologically quiescent clinically quiescent (SQCQ) patients and serologically active clinically active (SACA) patients served as control groups. Data including demographics, initial symptoms, duration to SACQ, treatments before and after SACQ, and characteristics of the flare group were analyzed.Results:Of the 682 patients, 170 were SACQ patients (24.9%), 187 were SQCQ patients, and 325 were SACA patients (47.7%). SQCQ patients (38.61±15.08 years old) were older at study start than SACQ patients (38.61±15.08 years vs. 32.09±14.35 years, p<0.001), but there was no significant difference between that of SACQ and SACA patients. 56 of the 170 SACQ patients (32.9%) experienced flare. Corticosteroids (OR 1.317, 95% CI 1.131 to 1.534; p<0.001) was an independent risk factor for flare, while antimalarials (OR 0.265, 95% CI 0.118 to 0.599; p=0.001) and immunosuppressants (OR 0.316, 95% CI 0.149 to 0.670; p=0.003) were protective factors.Conclusion:About one third of SLE patients with SACQ experience flare, more than that of patients with SQCQ. Thus, approach to prevent relapse in SACQ patient is required. Maintenance therapy of hydroxychloroquine and immunosuppressant agents may be protective and beneficial treatment strategy in these patients need further investigation.References:[1]Gladman DD, Urowitz MB, Keystone EC. Serologically active clinically quiescent systemic lupus erythematosus: a discordance between clinical and serologic features. Am J Med 1979; 66:210-5.[2]Huang WN, Tso TK, Wu HC, Yang HF, Tsay GJ. Impaired phagocytosis of apoptotic cell material in serologically active clinically quiescent patients with systemic lupus erythematosis. Int J Rheum Dis 2016; 19:1310-6.[3]Steiman AJ, Gladman DD, Ibañez D, Urowitz MB. Prolonged serologically active clinically quiescent systemic lupus erythematosus: frequency and outcome. J Rheumatol 2010; 37:1822-7.Disclosure of Interests:None declared

BackgroundSurgery remains the mainstay of treatment for resectable stage III non-small cell lung cancer (NSCLC). The preliminary results from some pilot trials have shown that neoadjuvant immunotherapy in NSCLC is safe and tolerable.1 2Hypothesizing that neoadjuvant toripalimab (a humanized anti-PD-1 antibody) plus chemotherapy can improve the outcome in resectable NSCLC, we are conducting a randomized, double-blind, placebo-controlled, phase III study to evaluate the efficacy and safety of toripalimab plus platinum-based doublet chemotherapy as neoadjuvant/adjuvant therapy for patients with resectable stage III NSCLC.MethodsThis ongoing study enrolls patients aged 18–70 years with treatment-naïve, histopathologically confirmed resectable stage III NSCLC without EGFR mutation or ALK translocation, ECOG PS 0–1, and adequate organ function. Eligible subjects are randomized (1:1) into experimental or control group, to receive perioperative toripalimab 240 mg or placebo combined with chemotherapy for 4 cycle in total (Docetaxel 60–75 mg/m2 or Paclitaxel 175 mg/m2 with platinum [squamous histology] or Pemetrexed 500 mg/m2 with platinum [non-squamous histology]) every 3 weeks for three cycles followed by surgery, and one more cycle after surgery, then monotherapy of toripalimab 240 mg or placebo every 3 weeks up to 13 cycles is delivered. Adjuvant radiotherapy is allowed. Randomization is stratified by tumor stage(IIIA vs IIIB), pathological type (squamous vs non-squamous), PD-L1 expression (PD-L1≥1% vs <1% or not evaluable) and planned surgical procedure (pneumonectomy vs lobectomy). Radiographic response is assessed within 4–6 weeks after last dose of neoadjuvant therapy, at 30 days after surgery and every 12 weeks thereafter. Primary endpoints are major pathologic response (MPR) rate evaluated by blind independent central pathology review (BIPR-MPR) and event-free survival evaluated by investigator (INV-EFS). Secondary endpoints include pathologic complete response (pCR) rate evaluated by BIPR and investigators (BIPR-pCR and INV-pCR), disease-free survival (DFS), 2–3 years OS rate, OS, safety, and feasibility of surgery. Exploratory endpoints are potential correlations between biomarkers and efficacy. A stratified Cochran Mantel Haenszel method will be used to assess binary endpoints. A Kaplan-Meier method, a stratified log-rank test and a stratified Cox proportional hazards model will be used to assess survival endpoints.Planned enrollment is 406 patients. The study is actively enrolling at 52 Chinese sites.ResultsN/AConclusionsN/AAcknowledgementsN/ATrial RegistrationThe Clinical trials. gov no NCT04158440Ethics ApprovalThis study was approved by the Ethics Board of all the involved sites; Approval number of Shanghai Chest Hospital: LS1936ConsentN/AReferencesForde PM, Chaft JE, Smith KN, et al. Neoadjuvant PD-1 blockade in resectable lung cancer N Engl J Med 2018;378:1976–1986Hellmann MD, Chaft JE, William WN Jr, et al. Pathological response after neoadjuvant chemotherapy in resectable non-small-cell lung cancers: proposal for the use of major pathological response as a surrogate endpoint. Lancet oncol 2014;15:e42–50.

BackgroundBackground: Early stage non-small cell lung cancer (NSCLC) possessed highly local and distant recurrence rates after surgery. Recent studies showed neoadjuvant therapy can improve pathological response and postoperative survival.1–3 Using immune checkpoint inhibitors as a neoadjuvant treatment could release neoantigens from dying tumour cells and stimulate the priming and expansion of neoantigen-specific T cells in the tumour before surgical resection.4 Previous study indicated radiotherapy combined with chemotherapy as neoadjuvant didn’t improve event-free or overall survival in stage III/N2 NSCLC.5 Subsequently, the study of neoadjuvant chemoradiation plus pembrolizumab have demonstrated that radiotherapy can enhance the effect of immunotherapy, achieving better complete pathological response.6Stereotactic body radiation therapy (SBRT) is a precise radiotherapy model that has shown good efficacy in treating early lung cancer, combining with immunetherapy as neoadjuvant therapy for NSCLC may improve pathological response and postoperative survival.MethodsThis is a prospective, single-center, two-part, phase II study assessing the safety, tolerability and efficacy of SBRT combined with toripalimab as neoadjuvant in patients with stage IIB–IIIA NSCLC without driver mutations (Clinical trial information: ChiCTR2000029277). Eligibility criteria include IIB–IIIA NSCLC (AJCC v8), adequate organ function, and ECOG PS 0 or 1. The primary endpoints were the safety and pathologic response, while the secondary endpoint was the radiographic response, recurrence-free survival and overall survival. The trial also aimed at exploring prognosis biomarkers (included Tumor infiltrates lymphocytes, CD8+Tcell, PD-L1 protein expression, tTMB and the correlation with pathologic response rate). 30 patients will receive SBRT (50 Gy/5 fractions over 5 days)on the first week. Afterwards, all patients will receive 2 cycles of intravenous toripalimab starting on Day15 (240 mg, q3w). Preoperative assessment will be performed two weeks (week 8) before surgery (week 10). In part 1 of the study, 6 patients will be enrolled to determine the safety and feasibility of the combination therapy. Subjects will be followed up for 90 days to observe perioperative adverse events. If the rate of delayed surgery (delay time >37 days) is >90% or the rate of grade 3/4 adverse effects is >70%, the study will be terminated. if not, 24 patients will be enrolled in part 2 of the study. This study is currently open and accruing patients.ReferencesCascone T, William WN Jr, Weissferdt A, et al. Neoadjuvant nivolumab or nivolumab plus ipilimumab in operable non-small cell lung cancer: the phase 2 randomized NEOSTAR trial. Nat Med 2021;27(3):504–514.Provencio M, Nadal E, Insa A, et al. Neoadjuvant chemotherapy and nivolumab in resectable non-small-cell lung cancer (NADIM): an open-label, multicentre, single-arm, phase 2 trial. Lancet Oncol 2020 Nov;21(11):1413–1422.Forde PM, Chaft JE, Smith KN, et al. Neoadjuvant PD-1 Blockade in Resectable Lung Cancer. N Engl J Med 2018 May 24;378(21):1976–1986.Liu J, Blake SJ, Yong MC, et al. Improved efficacy of neoadjuvant compared to adjuvant immunotherapy to eradicate metastatic disease. Cancer Discov 2016;6: 1382–99.Pless M, Stupp R, Ris HB, et al. Induction chemoradiation in stage IIIA/N2 non-small-cell lung cancer: a phase 3 randomised trial. Lancet 2015 12;386(9998): 1049–56.Lemmon C, Videtic GM. M, Murthy SC, et al. A phase I safety and feasibility study of neoadjuvant chemoradiation plus pembrolizumab followed by consolidation pembrolizumab in resectable stage IIIA non-small cell lung cancer. J Clin Oncol 2020; 38 (suppl; abstr 9009).

Background75% of diagnosed bladder tumors are non-muscle-invasive (NMIBC)[1, 2]. Most require intravesical instillation of M.bovis Bacillus Calmette-Guérin (BCG). Recurrence after immunotherapy occurs in ~50% patients. Development of treatments for BCG-resistant disease has lagged partly because few studies have attempted to understand the relationship between timing of tumor recurrence, reasoning for the recurrence, and the state of immune system at the time of recurrence.Immune exhaustion is observed following microbial infections, cancers and chronic inflammation [3–5]. Natural Killer (NK) cells are among the earliest responders[6–8] and undergo a similar program of exhaustion as T cells[9]. HLA-E strongly inhibits NKG2A-expressing NK and CD8+T cells and is commonly upregulated on tumors[10]. We evaluated the potential restorative capacity of NKG2A and PD-L1-blockade for reinvigorating NK and CD8+T cell antitumor functions in in BCG-resistant bladder cancer.Methods mRNA analysis of 2,892 genes was performed on tumor tissue of NMIBC patients before and after BCG therapy (n=35). Immunostaining (serial-IHC,immunofluorescence,imaging-mass cytometry) was performed on consecutive tissue sections. Single-cell-RNA-sequencing (scRNAseq) was performed on fresh bladder tumors (NMIBC,n=4; MIBC,n=9). OLink Proteomics (”Inflammation” panel) was performed longitudinally on plasma/urine from a prospective cohort of NMIBC patients. Patient tumors (n=3) were expanded as organoids and co-cultured with autologous tumor-derived NK and CD8+T cells in presence/absence of anti-PD-L1/NKG2A antibodies.ResultsWe demonstrate a robust local TME and systemic response to BCG that correlates with chronic inflammation and adaptive resistance rather than with preventing tumor recurrence. This resistance is mediated through IFN-γ-production by tumor-infiltrating NKG2A+NK and NKG2A+PD-1+CD8+T cells and results in increased HLA-E and PD-L1 on recurring tumors. Co-culture of treatment-naïve NMIBC tumors with recombinant IFN-gamma directly enhanced expression of PD-L1 and HLA-E. Longitudinal analysis of plasma before and during BCG immunotherapy revealed an inflammatory signature, including but not limited to IFN-gamma, that is maintained throughout treatment.Immunostaining and scRNAseq of NMIBC specimens revealed highly enriched infiltration by NKG2A+NK and NKG2A+CD8+T cells in HLA-EBrightPD-L1+ tumors and were spatially organized relative to tumors in a manner suggesting direct inhibition. Tumor-derived NK and CD8+T cells from BCG-resistant patients were co-cultured with autologous tumor organoids. Preliminary analyses demonstrated an improved anti-tumor response in presence of NKG2A/PD-L1-blockade.ConclusionsOur data support a model of BCG-resistance that points to a novel checkpoint axis that contributes to BCG-resistance: HLA-E/NKG2A. New insights into this axis in NMIBC and how it is altered with repeated BCG exposure will enable us to explore combination therapies (PD-L1/NKG2A-blockade) that may reduce BCG-resistance and provide durable response.ReferencesEidinger D, Morales A: Discussion paper: treatment of superficial bladder cancer in man. Ann N Y Acad Sci 1976, 277:239–240.Morales A, Eidinger D, Bruce AW: Intracavitary Bacillus Calmette-Guerin in the treatment of superficial bladder tumors. J Urol 1976, 116:180–183.Blank CU, Haining WN, Held W, Hogan PG, Kallies A, Lugli E, Lynn RC, Philip M, Rao A, Restifo NP et al: Defining ‘T cell exhaustion’. Nat Rev Immunol 2019, 19:665–674.Hashimoto M, Kamphorst AO, Im SJ, Kissick HT, Pillai RN, Ramalingam SS, Araki K, Ahmed R: CD8 T Cell Exhaustion in Chronic Infection and Cancer: Opportunities for Interventions. Annu Rev Med 2018, 69:301–318.McLane LM, Abdel-Hakeem MS, Wherry EJ: CD8 T Cell Exhaustion During Chronic Viral Infection and Cancer. Annu Rev Immunol 2019, 37:457–495.Lanier LL: NK cell receptors. Annu Rev Immunol 1998, 16:359–393.Biron CA, Gazzinelli RT: Effects of IL-12 on immune responses to microbial infections: a key mediator in regulating disease outcome. Curr Opin Immunol 1995, 7:485–496.Welsh RM, Jr.: Cytotoxic cells induced during lymphocytic choriomeningitis virus infection of mice. I. Characterization of natural killer cell induction. J Exp Med 1978, 148:163–181.da Silva IP, Gallois A, Jimenez-Baranda S, Khan S, Anderson AC, Kuchroo VK, Osman I, Bhardwaj N: Reversal of NK-cell exhaustion in advanced melanoma by Tim-3 blockade. Cancer Immunol Res 2014, 2:410–422.van Hall T, Andre P, Horowitz A, Ruan DF, Borst L, Zerbib R, Narni-Mancinelli E, van der Burg SH, Vivier E: Monalizumab: inhibiting the novel immune checkpoint NKG2A. J Immunother Cancer 2019, 7:263.Ethics ApprovalPrimary urothelial bladder cancer tumor tissue was obtained after obtaining informed consent in the context of an Institutional Review Board (IRB)-approved genitourinary cancer clinical database and specimen collection protocol (IRB #10-1180) at the Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai (New York, NY).

The toxicity of ammonia nitrogen (AN) has always caused severe harm to aquatic animals in intensive aquaculture conditions, especially in saline-alkali aquaculture waters. The application of AN removal bacteria is a safe and effective method for controlling the AN concentration in aquaculture water through direct conversion to bacterial protein. However, there is still a lack of AN removal bacteria that are appropriate for saline-alkali aquaculture conditions. In this study, three AN removal strains, namely, Bacillus idriensis CT-WN-B3, Bacillus australimaris CT-WL5-10, and Pseudomonas oleovorans CT-WL5-6, were screened out under alkaline conditions from the alkali-tolerant strains distributed in carbonate saline-alkali soil and water environments in Northeast China. Under different pH (8.0–9.0), salinities (10–30 g/L NaCl), alkalinities (10–30 mmol/L NaHCO3), and AN concentrations (1–3 mg/L), corresponding to the actual conditions of saline-alkali aquaculture waters, the AN removal rates and relative characteristics of these strains were analyzed. The results showed that all of the three strains were efficient on AN removal under various conditions, and the highest removal rate reached up to 3 × 10–13 mg/cfu/h. Both CT-WL5-10 and CT-WL5-6 were most efficient under pH 9.0 with 3 mg/L initial AN, while pH 8.5 with 2 mg/L AN was the best fit for CT-WN-B3. In 96-h pure incubation of these strains in alkali media, approximately 90% AN was removed, and pH values were decreased by 2.0 units within 12 h accompanied by the growth of the strains. In addition, salinity and alkalinity slightly disturbed the removal rates of CT-WL5-10 and CT-WL5-6, but there were at least 65% AN removed by them within 24 h. These results indicated that all three strains have good application prospect in saline-alkali aquaculture waters.