Histidine-Mediated Nickel and Zinc Translocation in Intact Plants of the Hyperaccumulator Noccaea caerulescens

In this work, the effect of exogenous histidine supply on zinc (Zn) and nickel (Ni) translocation to the shoots in intact plants of the hyperaccumulator Noccaea caerulescens F.K. Mey was studied. Three series of experiments were carried out. (1) Intact N. caerulescens plants (St-Félix-de-Pallières population) were pretreated for 4 h (12:00 till 16:00) with a MES/KOH-buffered 1 mM L-histidine solution or demineralized water, then exposed overnight (20 h) to 5, 25 or 250 µM Ni or Zn and harvested. (2) Intact N. caerulescens plants of the same population were pretreated with 1 mM L-histidine solution or demineralized water overnight (20 h) and then exposed to 250 µM Ni or Zn for 8 h during the day (10:00 till 18:00) and harvested. (3) Intact N. caerulescens plants (the calamine populations St-Félix-de-Pallières (SF) and La Calamine (LC), and the ultramafic population Monte Prinzera (MP)) were exposed for 8 h (10:00 till 18:00) to 250 µM Ni or Zn and then to 1 mM L-histidine solution or demineralized water overnight (20 h) and harvested. The Ni and Zn concentrations in the roots and shoots were determined by atomic absorption spectrophotometry. The translocation factor (TF), expressed as the shoot to root metal concentration ratio, the total plant Ni or Zn content, and the percentage of the total Ni or Zn content present in the shoot (% translocated) were calculated. A 4 h pretreatment with L-histidine during the afternoon (before metal exposure overnight) significantly decreased the Ni and Zn concentrations in the root and increased the concentration of Ni, but not of Zn, in the shoot, significantly increased both TF and the % translocated for both metals, albeit much more strongly for Ni, and also slightly, but significantly, increased the total plant content of Ni, but not of Zn. Overnight pretreatment with L-histidine (followed by metal exposure during the day) of the same population (SF) had basically similar effects on Ni translocation, but significantly decreased the plant total Ni content, and was without significant effects on Zn translocation, but considerably decreased the root Zn concentration. The different populations under study (SF, MP, LC) showed significant differences in their Ni and Zn uptake and translocation capacities, but in general showed qualitatively similar responses to post-treatment with L‑histidine that strongly increased the TF and the % translocated for both metals in SF and MP, whereas in LC the effect was prominent only for Ni. Significant population × histidine treatment effect interactions were obtained for the root Zn concentration, and the TF and % translocated for Ni, which were largely explained by a relatively low responsiveness to the L-histidine treatment in LC, compared to SF and/or MP. It is concluded that the high endogenous L-histidine concentrations in N. caerulescens are probably functional in the hyperaccumulation of both Ni and Zn. The overall stronger effect of exogenous L-histidine supply on the translocation of Ni, compared to Zn, seems to result, at least in part, from the high Zn burdens at the start of the treatments, particularly in the shoots, which largely mask the apparent effects of exogenous L-histidine supply on the shoot Zn concentration and, to a lower degree, the % Zn translocated

Non-immunogenic recombinant staphylokinase versus alteplase for patients with acute ischaemic stroke 4·5 h after symptom onset in Russia (FRIDA): a randomised, open label, multicentre, parallel-group, non-inferiority trial

Background: Non-immunogenic staphylokinase is modified recombinant staphylokinase with low immunogenicity, high thrombolytic activity, and selectivity to fibrin. We aimed to assess the safety and efficacy of a single intravenous bolus of non-immunogenic staphylokinase compared with alteplase in patients with acute ischaemic stroke within 4·5 h after symptom onset. Methods: We did a randomised, open-label, multicentre, parallel-group, non-inferiority trial in 18 clinical sites in Russia. We included patients aged 18 years and older with a diagnosis of acute ischaemic stroke (up to 25 points on the National Institutes of Health Stroke Scale). The study drug had to be administered within 4·5 h after the onset of symptoms. Patients were randomly assigned to receive either non-immunogenic staphylokinase (10 mg) or alteplase (0·9 mg/kg, maximum 90 mg), both administered intravenously. The randomisation sequence was created by an independent biostatistician using computer-generated random numbers. 84 blocks (block size of four) of opaque sealed envelopes were numbered sequentially from 1 to 336 and were opened in numerical order. Patients were unaware of their assigned treatment and were assessed by the study investigators who were also unaware of the treatment assignment on all trial days. Emergency department staff, who administered the assigned drug and opened the envelopes, were not masked to treatment. The primary efficacy endpoint was a favourable outcome, defined as a modified Rankin scale (mRS) score of 0–1 on day 90. The margin of non-inferiority was established as 16% for the difference in mRS score of 0–1 on day 90. Non-inferiority was tested using Welch's t-test for the primary outcome only. Endpoints were analysed in the per-protocol population, which comprised all randomly assigned patients who completed treatment without any protocol violations; this population was identical to the intention-to-treat population. This trial is completed and registered at ClinicalTrials.gov, NCT03151993. Findings: Of 385 patients recruited from March 18, 2017, to March 23, 2019, 336 (87%) were included in the trial. 168 (50%) patients were randomly assigned to receive non-immunogenic staphylokinase and 168 (50%) to receive alteplase. The median duration of follow-up was 89 days (IQR 89–89). 84 (50%) of 168 patients in the non-immunogenic staphylokinase group had a favourable outcome at day 90 compared with 68 (40%) of 168 patients in the alteplase group (odds ratio [OR] 1·47, 95% CI 0·93 to 2·32; p=0·10). The difference in the rate of favourable outcome at day 90 was 9·5% (95% CI –1·7 to 20·7) and the lower limit did not cross the margin of non-inferiority (pnon-inferiority <0·0001). Symptomatic intracranial haemorrhage occurred in five (3%) patients in the non-immunogenic staphylokinase group and in 13 (8%) patients in the alteplase group (p=0·087). On day 90, 17 (10%) patients in the non-immunogenic staphylokinase group and 24 (14%) patients in the alteplase group had died (p=0·32). 22 (13%) patients in the non-immunogenic staphylokinase group had serious adverse events, compared with 37 (22%) patients in the alteplase group (p=0·044). Interpretation: Non-immunogenic staphylokinase was non-inferior to alteplase for patients with acute ischaemic stroke. Mortality, symptomatic intracranial haemorrhage, and serious adverse events did not differ significantly between groups. Future studies are needed to continue to assess the safety and efficacy of non-immunogenic staphylokinase in patients with acute ischaemic stroke within the 4·5 h time window, and to assess the drug in patients with acute ischaemic stroke outside this time window with reperfusion CT or magnetic resonance angiography followed by thrombectomy if necessary. Funding: The Russian Academy of Sciences. © 2021 Elsevier Lt