Control of nitrification of fertilizer nitrogen: Effect of inhibitors,banding and nesting

Laboratory incubation and field experiments were conducted to evaluate thiourea, ATC (4-amino-1, 2, 4 triazole hydrochloride) and N-Serve 24 E (2-chloro-6-trichloromethyl-pyridine) as inhibitors of nitrification of fertilizer N. In the incubation experiment, most of the added aqueous NH₃ or urea was nitrified at 14 days on both soils, but addition of the inhibitors to fertilizer N decreased the conversion of NH₄−N to NO₃−N markedly. There was less nitrification for ATC and thiourea but not for N-Serve 24 E when the fertilizers and the inhibitors were placed at a point as opposed to when mixed into soil. After 28 days, ATC and N-Serve 24 E were more effective in inhibiting nitrification than thiourea. ATC and N-Serve 24 E also inhibited release of mineral N (NH₄−N+NO₃−N) from native soil N. In the uncropped field experiment, which received N fertilizers in the fall, nitrification of fall-applied N placed in the 15-cm bands was almost complete by early May in the Malmo soil, but not in the Breton soil. When ATC or thiourea had been applied with urea, nitrification of fall-applied N was depressed by May and the recovery of applied N as NH₄−N was greater with increasing band spacing to 60 cm or placing N fertilizer in nests (a method of application where urea prills were placed at a point in the soil in the center of 60×60 cm area). In late June, the percentage recovery of fall-applied N in soil as NH₄−N or mineral N increased with wide band spacing, or nest placement, or by adding ATC to fertilizer N on both soils. These results indicate that placing ammonium-based N fertilizers in widely-spaced bands or in nests with low rates of inhibitors slows nitrification enough to prevent much of the losses from fall-applied N. Scientific Paper No. 552, Lacombe Research Station, Research Branch, Agric, Can.     

Overexpression of RYBP inhibits proliferation,invasion, and chemoresistance to cisplatin in anaplastic thyroid cancer cells via the EGFR pathway

Ring1 and YY1 binding protein (RYBP), a new member of the polycomb group protein family, has been reported to play an important role in various biological processes. Recently, more and more studies have demonstrated an implication of RYBP in cancer development. However, the specific role of RYBP in anaplastic thyroid cancer (ATC) remains unknown. In this study, we investigated for the first time the expression pattern and biological functions of RYBP in ATC. We showed that RYBP was lowly expressed in ATC tissues and cell lines. We also found that overexpression of RYBP inhibited ATC cell proliferation, invasion, and cisplatin resistance. Furthermore, we observed that upregulation of RYBP decreased the phosphorylation of EGFR and ERK1/2 in ATC cells. Taken together, our data indicated that RYBP might be considered as a promising therapeutic target for the treatment of ATC.     

凝血与纤溶指标与颅脑损伤后急性创伤性凝血病的关系及对脑心综合征的预测效能研究

摘要 目的：分析凝血与纤溶指标与颅脑损伤（TBI）后急性创伤性凝血病（ATC）的关系及对脑心综合征（CCS）的预测效能。方法：选取2020年1月～2022年12月本院收治的80例TBI患者作为研究对象,分别检测并计算非ATC患者与ATC患者、非CCS患者与CCS患者的凝血酶原时间（PT）、活化部分凝血酶原时间（APTT）、D-二聚体（D-D）、凝血酶-抗凝血酶复合物（TAT）、纤溶酶-α2纤溶酶抑制物复合物（PIC）水平和国际标准化比值（INR）、TAT/PIC比值;并采用双变量Spearman相关性检验凝血与纤溶指标的相关性,建立多因素Logistic模型分析TBI后ATC和TBI合并CCS的影响因素,同时分析凝血与纤溶指标对TBI合并CCS的预测效能。结果：80例TBI患者的ATC发生率为27.50%;与非ATC组比较,ATC组PT、APTT、D-D水平较高,TAT/PIC比值较低（P＜0.05）。PT、APTT、D-D与颅脑损伤后ATC呈正相关性,TAT/PIC与ATC呈负相关性。多因素logistic分析结果显示,TAT/PIC是颅脑损伤后ATC的独立危险因素（P＜0.05）。80例TBI患者的CCS发生率为20.00%;与非CCS组比较,CCS组PT、APTT、D-D水平较高,TAT/PIC比值较低（P＜0.05）。多因素logistic分析结果显示,PT、APTT、D-D、TAT/PIC是TBI合并CCS的独立危险因素（P＜0.05）。TBI合并CCS预测中,PT、APTT、D-D的ACU均＞0.70,TAT/PIC＞0.85。结论：TAT/PIC与TBI后ATC存在一定关联,同时还能预测CCS的发生,在TBI患者预后预测方面具有指导意义。     

Enhanced dissolution of the substrate d,l‐2‐amino‐Δ2‐thiazoline‐4‐carboxylic acid and enzymatic production of l‐cysteine at high concentrations

l ‐Cysteine is widely used as a precursor in the pharmaceutical, cosmetic, food, and feed additive industries. It has been industrially produced from hydrolysis of human and animal hairs, which is limited for industrial production. At the same time, chemical hydrolysis causes the formation of intractable waste material. Thus, environmentally friendly methods have been developed. A big obstacle of currently available methods is the low substrate solubility leading to poor l ‐cysteine yield. Here, a method for improving the low solubility of the substrate d ,l ‐2‐amino‐Δ²‐thiazoline‐4‐carboxylic acid (d ,l ‐ATC) is presented and the enzymatic reaction at high concentration levels was optimized. The substrate was dissolved in large amounts in aqueous solutions by pH control using salts. d ,l ‐ATC solubility increased with an increasing solution pH due to its enhanced hydrophilicity, which can be achieved by a shift to dissociated carboxylic group (–COO^?). The highest d ,l ‐ATC solubility of 610 mM was obtained at pH 10.5. The maximum l ‐cysteine yield of 250 mM was attained at pH 9.1, which lies between the optimum values for high substrate solubility and reaction rate. The product yield could be increased by more than 10 times compared to those in previous reports, which is industrially meaningful.     

The crystal structures of ornithine carbamoyltransferase from Mycobacterium tuberculosis and its ternary complex with carbamoyl phosphate and L-norvaline reveal the enzyme's catalytic mechanism

Mycobacterium tuberculosis ornithine carbamoyltransferase (Mtb OTC) catalyzes the sixth step in arginine biosynthesis; it produces citrulline from carbamoyl phosphate (CP) and ornithine (ORN). Here, we report the crystal structures of Mtb OTC in orthorhombic (form I) and hexagonal (form II) space groups. The molecules in form II are complexed with CP and l-norvaline (NVA); the latter is a competitive inhibitor of OTC. The asymmetric unit in form I contains a pseudo hexamer with 32 point group symmetry. The CP and NVA in form II induce a remarkable conformational change in the 80s and the 240s loops with the displacement of these loops towards the active site. The displacement of these loops is strikingly different from that seen in other OTC structures. In addition, the ligands induce a domain closure of 4.4° in form II. Sequence comparison of active-site residues of Mtb OTC with several other OTCs of known structure reveals that they are virtually identical. The interactions involving the active-site residues of Mtb OTC with CP and NVA and a modeling study of ORN in the form II structure strongly rule out an earlier proposed mechanistic role of Cys264 in catalysis and suggest a possible mechanism for OTC. Our results strongly support the view that ORN with an already deprotonated N^ε atom is the species that binds to the enzyme and that one of the phosphate oxygen atoms of CP is likely to be involved in accepting a proton from the doubly protonated N^ε atom of ORN. We have interpreted this deprotonation as part of the collapse of the transition state of the reaction.     

Inhalation toxicity of soman vapor in non-anesthetized rats: A preliminary assessment of inhaled bronchodilator or steroid therapy

Respiratory toxicity, injury and treatment following vapor inhalational exposure to the chemical warfare nerve agent (CWNA) soman (GD) were examined in non-anesthetized rats. This study exposed male Sprague–Dawley rats (250–300 g) to 520, 560, 600, 825 or 1410 mg × min/m³ of soman in a customized head-out inhalation system. Signs of CWNA-induced cholinergic crises were observed in all soman-exposed animals. The LCt50 of vaporized soman as determined by probit analysis was 593.1 mg × min/m³. All animals exposed to 825 and 1410 mg × min/m³ developed severe convulsions and died within 4–8 min post-exposure. Edema measured by wet/dry weight ratio of the left lung lobe increased in a dose-dependent manner in all soman-exposed animals. Bronchoalveolar lavage (BAL) fluid and blood acetylcholinesterase (AChE) activities were inhibited dose-dependently in soman-exposed groups at 24 h. A significant increase in total BAL protein was observed in soman-exposed animals at all doses. AChE activity was inhibited in lung and whole brain tissues in all soman-exposed animals. Histopathological analysis of the lungs of animals exposed to 600 mg × min/m³ of soman revealed prominent morphological changes including alveolar histiocytosis, hemorrhage and inflammation consisting of neutrophilic exudate. Exposure of animals to 600 mg × min/m³ of soman followed by treatment with two actuations for 10 s of Combivent (21 μg of ipratropium bromide and 120 μg of albuterol sulfate) and Symbicort (80 μg budesonide and 4.5 μg formoterol) by inhalation into a modified metered dose inhaler (MDI) 10 min post-exposure resulted in increased minute volume, but did not decrease mortality. These results indicate that inhalation exposure to soman vapor causes acute respiratory toxicity and injury in untreated, un-anesthetized rats and that inhalation treatment with Combivent or Symbicort did improve the respiratory outcomes, but did not influence lethality.     

Arabidopsis thaliana CENTRORADIALIS homologue (ATC) acts systemically to inhibit floral initiation in Arabidopsis

Floral initiation is orchestrated by systemic floral activators and inhibitors. This remote‐control system may integrate environmental cues to modulate floral initiation. Recently, FLOWERING LOCUS T (FT) was found to be a florigen. However, the identity of systemic floral inhibitor or anti‐florigen remains to be elucidated. Here we show that Arabidopsis thaliana CENTRORADIALIS homologue (ATC), an Arabidopsis FT homologue, may act in a non‐cell autonomous manner to inhibit floral initiation. Analysis of the ATC null mutant revealed that ATC is a short‐day‐induced floral inhibitor. Cell type‐specific expression showed that companion cells and apex that express ATC are sufficient to inhibit floral initiation. Histochemical analysis showed that the promoter activity of ATC was mainly found in vasculature but under the detection limit in apex, a finding that suggests that ATC may move from the vasculature to the apex to influence flowering. Consistent with this notion, Arabidopsis seedling grafting experiments demonstrated that ATC moved over a long distance and that floral inhibition by ATC is graft transmissible. ATC probably antagonizes FT activity, because both ATC and FT interact with FD and affect the same downstream meristem identity genes APETALA1, in an opposite manner. Thus, photoperiodic variations may trigger functionally opposite FT homologues to systemically influence floral initiation.     

Effect of ATC,N-Serve 24 E and thiourea nitrification inhibitors on yield and N uptake of barley fertilized with fall-applied N1

Solution urea and aqua NH₃ were injected in bands 9 cm deep and spaced 45 cm apart with and without nitrification inhibitors during October in 10 field experiments in north-central Alberta. ATC (4-amino-1,2,4-triazole hydrochloride), N-Serve 24 E (2-chloro-6-trichloromethyl-pyridine) and thiourea were used in two experiments, ATC only in another six experiments, and N-Serve 24 E only in another two experiments. Yield and apparent recovery of applied N in barley grain were determined. In the two experiments where fall treated plots were soil sampled in the following spring, 44% of the fall-applied N was recovered in the soil when inhibitors were not used. But where the inhibitors were added to the fall-applied N as NH₄−N in May was 4% and 31% without and with addition of inhibitors, respectively. Likewise, in experiments where three inhibitors were used, the treatments with inhibitors increased the yield and N recovery in grain by more than 50% compared to fall application without inhibitors. In the other experiments, fall-applied ATC or N-Serve 24 E did not always increase yield or N recovery in grain. Considering all experiments with ATC, the average recovery of applied N in barley grain was 28, 40 and 57% for fall banding, fall banding with ATC and spring application, respectively. In view of this and previous work in north-central Alberta, inhibitors injected in bands in the fall slowed nitrification and improved yield, but nests or large granules of urea were more effective. Scientific paper No 553, Lacomba Research Station, Research Branch, Agriculture Canada.     

Design,synthesis and biological evaluation of phthalimide-alkylamine derivatives as balanced multifunctional cholinesterase and monoamine oxidase-B inhibitors for the treatment of Alzheimer’s disease

A series of novel phthalimide-alkylamine derivatives were synthesized and evaluated as multi-functions inhibitors for the treatment of Alzheimer’s disease (AD). The results showed that compound TM-9 could be regarded as a balanced multi-targets active molecule. It exhibited potent and balanced inhibitory activities against ChE and MAO-B (huAChE, huBuChE, and huMAO-B with IC₅₀ values of 1.2 μM, 3.8 μM and 2.6?μM, respectively) with low selectivity. Both kinetic analysis of AChE inhibition and molecular modeling study suggested that TM-9 binds simultaneously to the catalytic active site and peripheral anionic site of AChE. Interestingly, compound TM-9 abided by Lipinski’s rule of five. Furthermore, our investigation proved that TM-9 indicated weak cytotoxicity, and it could cross the blood-brain barrier (BBB) in vitro. The results suggest that compound TM-9, an interesting multi-targeted active molecule, offers an attractive starting point for further lead optimization in the drug-discovery process against Alzheimer’s disease.