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Evaluation of variability of silymarin complex in Silybi mariani fructus harvested during two production years 两个生产年份水飞蓟果实水飞蓟素复合物变异性的评价
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2020-0023
M. Habán, D. Zvercová, M. Adamjaková
Abstract Milk thistle [Silybum marianum (L.) Gaertn.], a member of Asteraceae family, is one of the most cultivated medicinal plants widespread throughout the world. The pharmacological drug is a ripe fruit without pappus – Silybi mariani fructus – containing flavonolignans and generating silymarin complex. In folk medicine, it is used for the treatment of liver disorders, kidney problems, rheumatism as well as gastronomic disturbances, cardiac and neurotic disorders, and fever. The components of silymarin complex are useful in cancer prevention and treatment. The aim of the study was to determine the amount of silymarin complex contained in the fruit of the harvest of two consecutive years and how much they differ from one another. Representative samples of fruit were collected in 2015 and 2016 and distributed by a company Agrofos (Slovakia). Regarding the analytical method, we used a high-performance liquid chromatography (HPLC); the method was approved by the European Pharmacopoeia 10. The statistical significance was on the level P < 0.05. The total content of silymarin complex was 15.28 ± 0.06 g.kg−1 (in 2015) and 16.65 ± 0.09 g.kg−1 (in 2016). In both studied years, the highest representation of silybin B was observed (7.04 ± 0.07 g.kg−1 versus 5.92 ± 0.08 g.kg−1). The differences between the individual fractions of the silymarin complex were statistically significant. There was also a significant difference of 9% in the total silymarin content between 2015 and 2016. In conclusion, we can state that both samples of Silybi mariani fructus meet the requirements of the European Pharmacopoeia.
摘要乳蓟(Silybum marianum(L.)Gaertn.)是菊科的一员,是世界上种植最广泛的药用植物之一。该药物是一种不含pappus的成熟水果——水飞蓟果——含有黄酮类化合物并产生水飞蓟素复合物。在民间医学中,它被用于治疗肝脏疾病、肾脏问题、风湿病以及胃肠道疾病、心脏和神经系统疾病以及发烧。水飞蓟素复合物的组分可用于癌症的预防和治疗。该研究的目的是确定连续两年收获的果实中所含水飞蓟素复合物的含量,以及它们之间的差异。2015年和2016年采集了具有代表性的水果样本,并由Agrofos(斯洛伐克)公司分发。关于分析方法,我们使用了高效液相色谱法;该方法被欧洲药典10批准。差异有统计学意义(P<0.05)。水飞蓟素复合物的总含量为15.28±0.06 g.kg-1(2015年)和16.65±0.09 g.kg-1(2016年)。在两个研究年份中,观察到水飞蓟宾B的最高代表性(7.04±0.07 g.kg−1对5.92±0.08 g.kg−2)。水飞蓟素复合物的各个部分之间的差异具有统计学意义。2015年至2016年间,水飞蓟素总含量也存在9%的显著差异。总之,我们可以声明水飞蓟果的两个样品都符合欧洲药典的要求。
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引用次数: 0
Book of Abstracts 38th Technology Days 9th and 10th September 2021 第38届技术日2021年9月9日和10日
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2021-0010
M. Molitorisová
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引用次数: 0
Nephroprotective Effect of Coenzyme Q10 alone and in Combination with N-acetylcysteine in Diabetic Nephropathy 辅酶Q10单独及联合N-乙酰半胱氨酸对糖尿病肾病的保护作用
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2020-0020
Manoj Mahajan, A. Upaganlawar, Chandrashekar D. Upasani
Abstract Aim Oxidative stress due to chronic hyperglycaemia is a key factor in the development and progression of various microvascular complications including diabetic nephropathy (DN) and associated renal injury. Treatment with antioxidants is one of the strategies to protect the kidney from oxidative tissue damage to improve renal physiology during DN. The investigation, therefore, was designed to assess the nephroprotective effect of coenzyme Q10 (CoQ10) and N-acetylcysteine (NAC), either alone or in combination in streptozotocin (STZ)-nicotinamide (NAD) induced diabetic nephropathy (DN) in rats. Methods T2DM induced by STZ (55 mg/kg, i.p.)-NAD (110 mg/kg, i.p.) in Sprague-Dawley rats (220–250 g) was confirmed by the elevated blood glucose level and glycated haemoglobin. DN was assessed by renal function tests. The diabetic rats were treated with CoQ10 (10 mg/kg, p.o.) and/or NAC (300 mg/kg, p.o.) for 8 weeks after confirmation of DN. Oxidative tissue damage due to STZ-NAD was estimated by malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT), reduced glutathione (GSH), myeloperoxidase (MPO) and nitric oxide (NO) in the renal homogenate. Results Data showed significant alteration in serum and urinary creatinine, total protein, albumin, serum urea, blood urea nitrogen (BUN) and uric acid in diabetic animals as compared to the control rats. CoQ10 and/or NAC effectively alleviated the disturbances in renal function. Diabetic rats showed increased MDA, decreased SOD and CAT activities and decreased GSH along with a significant increase in MPO activity and nitrite content. Treatment with the aforementioned antioxidants and their combination ameliorated the kidney damage as indicated by the reduced OS with improved renal function. Conclusion The investigation suggests that the chronic hyperglycaemia-induced OS leads to the development and progression of DN. The combined treatment with CoQ10 and NAC has shown a remarkable nephroprotective effect suggesting that combined antioxidant therapy with CoQ10 and NAC may be useful in the attenuation of DN.
目的慢性高血糖引起的氧化应激是糖尿病肾病(DN)及相关肾损伤等多种微血管并发症发生发展的关键因素。抗氧化剂治疗是保护肾脏免受氧化性组织损伤以改善肾脏生理机能的策略之一。因此,本研究旨在评估辅酶Q10 (CoQ10)和n -乙酰半胱氨酸(NAC)单独或联合使用对链脲霉素(STZ)-烟酰胺(NAD)诱导的大鼠糖尿病肾病(DN)的肾保护作用。方法采用STZ (55 mg/kg, i.p)-NAD (110 mg/kg, i.p)诱导Sprague-Dawley大鼠(220 ~ 250 g) T2DM,血糖和糖化血红蛋白升高。通过肾功能检查评估DN。确认糖尿病大鼠DN后,给予辅酶q10 (10 mg/kg, p.o.)和/或NAC (300 mg/kg, p.o.) 8周。通过肾脏匀浆中的丙二醛(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)、还原性谷胱甘肽(GSH)、髓过氧化物酶(MPO)和一氧化氮(NO)来评估STZ-NAD引起的氧化性组织损伤。结果糖尿病动物血清和尿肌酐、总蛋白、白蛋白、血清尿素、血尿素氮(BUN)、尿酸等指标与对照组相比均有显著变化。辅酶q10和/或NAC可有效缓解肾功能紊乱。糖尿病大鼠MDA升高,SOD和CAT活性降低,GSH降低,MPO活性和亚硝酸盐含量显著升高。用上述抗氧化剂及其联合治疗可以改善肾损伤,这表明OS降低,肾功能改善。结论慢性高血糖诱导的OS可导致DN的发生发展。CoQ10和NAC联合治疗显示出显著的肾保护作用,提示CoQ10和NAC联合抗氧化治疗可能有助于DN的衰减。
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引用次数: 0
Phospholipid bilayers in model membranes and drug delivery systems: from physics to pharmacy 磷脂双分子层在模型膜和药物传递系统:从物理学到药学
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2021-0008
D. Uhríková
Abstract Lipids spontaneously aggregate in an aqueous environment forming supramolecular structures of various architectures known as liquid crystalline mesophases. Their thermodynamic properties determined by dual polar/apolar nature coupled with the possibility to modulate the structural parameters, phase geometry and stability are challenging for applications in drug delivery systems. We review a few examples of functionality of lipid bilayers.
摘要脂质在水性环境中自发聚集,形成各种结构的超分子结构,称为液晶中间相。它们的热力学性质由双极性/非极性决定,再加上调节结构参数、相几何形状和稳定性的可能性,对药物递送系统的应用具有挑战性。我们回顾了脂质双层功能的几个例子。
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引用次数: 0
Purification of Murine Gammaherpesvirus 68 With Use of Differential Centrifugation 差速离心纯化小鼠γ疱疹病毒68
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2021-0009
R. Hódosi, E. Nováková, M. Šupolíková
Abstract The method for separation of viral particles in a concentrated form from the environment is called virus purification. Viruses are required to be purified for a range of studies in which it is necessary to distinguish the properties or structure of a virus from the host cells or culture media, including analysis of viral polypeptide structures and membrane glycoprotein function. Our objective was to purify murine gammaherpesvirus 68 (MHV-68, MuHV-4) using the centrifuge, equipment and other materials available in our laboratory. After infection of baby hamster kidney 21 (BHK-21) cells with MHV-68 with the multiplicity of infection (MI) of 0.01 and following virus multiplication, we repeatedly froze and thawed the cell culture to disrupt the cells and release the virus particles into the culture medium. We used low-speed centrifugation (3000 rpm at 4°C) to separate the viral particles from cell debris. Subsequently, we transferred the supernatant containing virus particles to a fresh centrifuge tube and centrifuged at a speed of 8000 rpm (8801 g) and 11,000 rpm (=16,639 g) and at 4°C. We tested different centrifugation durations of 2, 4, 6 and 8 hours. To evaluate the quality of the obtained purified MHV-68 virus by this method and compare it to purified MHV-68 sample acquired by conventional ultracentrifugation on sucrose cushion (30%, w/v), we used the SDS-PAGE separation method using a 4%–20% (w/v) and 6%–14% (w/v) gradient gel. We obtained the best results with 6-hour-long centrifugation at 11,000 rpm. In conclusion, we managed to optimise virus purification method using the equipment available in our laboratory and prepared purified MHV-68 virus in sufficient concentration for determination of MHV-68 virus proteins.
从环境中分离浓缩形式的病毒颗粒的方法称为病毒纯化。在一系列研究中,需要对病毒进行纯化,以区分病毒与宿主细胞或培养基的特性或结构,包括病毒多肽结构和膜糖蛋白功能的分析。我们的目的是利用离心机、设备和实验室提供的其他材料纯化小鼠γ疱疹病毒68 (MHV-68, MuHV-4)。在感染倍数(MI)为0.01的MHV-68感染幼鼠肾21 (BHK-21)细胞后,在病毒增殖后,反复冻融细胞培养液,破坏细胞,将病毒颗粒释放到培养基中。我们使用低速离心(3000 rpm, 4°C)将病毒颗粒从细胞碎片中分离出来。随后,我们将含有病毒颗粒的上清液转移到新鲜的离心管中,在4°C的条件下,以8801 g和11000 rpm (= 16639 g)的速度离心。我们测试了不同的离心时间为2、4、6和8小时。为了评价该方法纯化的MHV-68病毒的质量,并将其与常规在蔗糖缓冲液(30%,w/v)上超离心纯化的MHV-68样品进行比较,我们采用SDS-PAGE分离方法,采用4%-20% (w/v)和6%-14% (w/v)梯度凝胶分离。我们在11000 rpm下离心6小时获得了最好的结果。综上所述,我们利用实验室现有的设备优化了病毒纯化方法,并制备了足够浓度的纯化MHV-68病毒,用于检测MHV-68病毒蛋白。
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引用次数: 0
Characterization of bioactive substances MHGF-68 on tumour cell lines with LiveFlow In Vitro Technology LiveFlow体外技术对肿瘤细胞系活性物质MHGF-68的表征
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2020-0021
R. Hódosi, E. Nováková, K. Macková, M. Molitorisová, M. Šupolíková
Abstract As part of experimental research, growth factor-like substances associated with MHV-68, named MHGF-68, were discovered in our laboratory. MHGF-68 activity was manifested by the ability to alter cell morphology, that is, normal phenotype to transformed, resp. suppresses the transformed phenotype of tumour cells. The aim of the experiments was to monitor the effect of MHGF-68 on the change of the cell actin cytoskeleton in the tumour cell line Hepa1c1c7, as well as the normal cell line NIH3T3, and compare conventional stationary cultivation and dynamic cultivation conditions using a LiveFlow system (In Vitro Technologies). LiveFlow is an advanced system to test the impact of different compounds on the cell cultures, which allows simulation of in vivo conditions thanks to continuous flow of cultivation medium. MHGF-68 was prepared with the infection of BHK-21 cells with MHV-68 virus under non-permissive conditions (41°C). After dynamic cultivation with MHGF-68, we observed changes in morphology on Hepa1c1c7 cells. In cells cultured in a dynamic environment, we observed more pronounced changes in cell morphology in comparison with cells cultured statically. We observed no changes in the cytoskeletal structures in the NIH 3T3 cell line affected by MHGF-68 in both types of cultivation. The advantage of LiveFlow in comparison to in vivo testing is that the experiments performed in this system are less time and money consuming. Dynamic cultivation in the LiveFlow system is suitable for optimizing experiments before testing substances in vivo.
作为实验研究的一部分,我们在实验室发现了与MHV-68相关的生长因子样物质,命名为MHGF-68。MHGF-68的活性表现为改变细胞形态的能力,即从正常表型转变为转化型。抑制肿瘤细胞的转化表型。本实验旨在监测MHGF-68对肿瘤细胞系Hepa1c1c7和正常细胞系NIH3T3细胞肌动蛋白骨架变化的影响,并利用LiveFlow系统(in Vitro Technologies)比较常规固定培养和动态培养条件。LiveFlow是一种先进的系统,用于测试不同化合物对细胞培养的影响,由于培养基的连续流动,它可以模拟体内条件。用MHV-68病毒感染BHK-21细胞,在非许可条件下(41℃)制备MHGF-68。MHGF-68动态培养后,我们观察到Hepa1c1c7细胞形态的变化。在动态环境中培养的细胞中,我们观察到与静态培养的细胞相比,细胞形态发生了更明显的变化。我们观察到MHGF-68在两种培养方式下对NIH 3T3细胞系的细胞骨架结构没有影响。与体内测试相比,LiveFlow的优点是在该系统中进行的实验花费的时间和金钱更少。LiveFlow系统中的动态培养适合于在体内测试物质之前优化实验。
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引用次数: 1
Study of the solubilisation process of bacterial model membranes induced by DDAO DDAO诱导细菌模型膜溶解过程的研究
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2020-0019
K. Želinská, J. Gallová
Abstract Solubilisation of two bacterial model membranes induced by N,N-dimethyl-1-dodecanamine-N-oxide (DDAO) was studied. The first model membrane consisted of a mixture of palmitoyloleoylphosphatidylethanolamine (POPE) and palmitoyloleoylphosphatidylglycerol (POPG) in a molar ratio 0.6:0.4 mol/mol, and a second model membrane was enriched with tetraoleoylcardiolipin (TOCL) with a composition POPE-POPG-TOCL = 0.67:0.23:0.1 mol/mol/mol. Solubilisation of these model membranes was studied by static light scattering (nephelometry). Effective ratio Re (the amount of DDAO integrated into the bilayer to the amount of lipid) at different steps of the solubilisation process was determined. The molar partition coefficient of DDAO was calculated – in case of the POPE-POPG membrane, Kp = 5,300 ± 400, for the POPE-POPG-TOCL membrane, Kp = 6,500 ± 500.
摘要研究了N,N-二甲基-1-十二胺-N-氧化物(DDAO)对两种细菌模型膜的增溶作用。第一种模型膜由棕榈酰油酰基磷脂酰乙醇胺(POPE)和棕榈酰油酰基磷脂酰甘油(POPG)组成,摩尔比为0.6:0.4 mol/mol;第二种模型膜富含四油酰基心磷脂(TOCL),组成为POPE-POPG-TOCL = 0.67:0.23:0.1 mol/mol/mol。通过静态光散射(浊度法)研究了这些模型膜的增溶作用。测定了溶解过程中不同步骤的有效比Re(整合到双分子层中的DDAO的量与脂质的量)。计算了DDAO的摩尔分配系数,对于POPE-POPG膜,Kp = 5,300±400,对于POPE-POPG- tocl膜,Kp = 6,500±500。
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引用次数: 0
Antimicrobial activity and cytotoxicity of transition metal carboxylates derived from agaric acid 木耳酸过渡金属羧酸盐的抗菌活性和细胞毒性
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2020-0018
L. Habala, Ľ. Pašková, A. Bilková, F. Bilka, B. Oboňová, J. Valentová
Abstract Carboxylato-type transition metal complexes with agaric acid, a bioactive natural compound derived from citric acid, were prepared, and tested in vitro for their antimicrobial activity and cytotoxicity. The products as well as agaric acid itself are amphiphilic compounds containing a hydrophilic head (citric acid moiety) and a hydrophobic tail (non-polar alkyl chain). The putative composition of the carboxylates was assigned on grounds of elemental analysis, infrared (IR) and high-resolution mass spectra (HR-MS), as well as in analogy with known complexes containing the citrate moiety. The metal carboxylates showed interesting activity in several microbial strains, especially against S. aureus (vanadium complex; MIC = 0.05 mg/ml). They were also tested for their cytotoxic activity in hepatocytes, the highest activity having been found in the copper(II) and manganese(II) complexes. Further research based on these preliminary results is needed in order to evaluate the influence of parameters like stability of the metal complexes in solution on the bioactivity of the complexes.
摘要以柠檬酸为原料制备了具有生物活性的天然化合物——木耳酸与羧化型过渡金属配合物,并对其抗菌活性和细胞毒性进行了体外测试。产物以及木耳酸本身是两亲性化合物,包含亲水性头部(柠檬酸部分)和疏水性尾部(非极性烷基链)。基于元素分析、红外(IR)和高分辨率质谱(HR-MS),以及与含有柠檬酸盐部分的已知配合物类似,确定了羧酸盐的推定组成。金属羧酸盐在几种微生物菌株中显示出有趣的活性,尤其是对金黄色葡萄球菌(钒络合物;MIC=0.05mg/ml)。还测试了它们在肝细胞中的细胞毒性活性,在铜(II)和锰(II)复合物中发现了最高的活性。需要在这些初步结果的基础上进行进一步的研究,以评估金属络合物在溶液中的稳定性等参数对络合物生物活性的影响。
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引用次数: 1
Rapid and simple CZE-UV method for quality control of B1 and B6 vitamins in drugs and dietary supplements 快速简便的CZE-UV法用于药物和膳食补充剂中B1和B6维生素的质量控制
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2021-0002
M. Matušková, I. Čižmárová, P. Chaľová, P. Mikuš, J. Piešťanský
Abstract The application of hydrodynamically closed capillary zone electrophoresis combined with convenient ultraviolet (UV) detection allows fast, simple, environmentally friendly and cost-effective analysis of ions or ionisable molecules. This technique has been used to determine two selected B vitamins (thiamine, pyridoxine) in various drug formulations. The developed method was characterised by excellent validation parameters, such as linearity, precision, accuracy, limit of detection and limit of quantification. The total time of analysis was lower than 13.5 min. The results indicate that the method is suitable for implementation in routine quality control of selected B vitamins in pharmaceutical and food samples.
摘要流体动力学封闭毛细管区带电泳与方便的紫外线(UV)检测相结合的应用,可以快速、简单、环保且经济高效地分析离子或电离分子。该技术已被用于测定各种药物制剂中两种选定的B族维生素(硫胺素、吡哆醇)。所开发的方法具有良好的验证参数,如线性、精密度、准确度、检测限和定量限。分析总时间小于13.5分钟。结果表明,该方法适用于药品和食品样品中所选B族维生素的常规质量控制。
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引用次数: 0
Possibilities of microemulsion application in rectal administration of indomethacin 微乳应用于直肠给药吲哚美辛的可能性
Pub Date : 2021-01-01 DOI: 10.2478/afpuc-2021-0012
M. Špaglová, M. Čuchorová, M. Čierna, V. Mikušová, K. Bauerová, S. Poništ
Abstract Rectal administration is a suitable route of administration for drugs that are either very irritating to the intestine (e.g., indomethacin) or are more effective when the first-pass effect in the liver is circumvented. Microemulsions are a tool for the improvement of penetration of sparingly soluble drugs. They are mainly used in topical and transdermal drug delivery. However, they find application also in other routes of administration, mainly due to their ability to solubilize sparingly soluble drugs. The selection of a suppository base depends on the physical properties of the drug. The study focused on evaluating the effect of the microemulsion as the solubilizer of sparingly soluble indomethacin in hydrophilic and lipophilic suppository bases compared with Polysorbate 80 as the excipient contained in the microemulsion. The reference suppositories were prepared by the traditional moulding technique from Adeps solidus or Macrogol suppository base without the previous drug solubilization. The microemulsion-based suppositories were prepared after the initial solubilization of the drug in the microemulsion or Polysorbate 80, followed by the addition of suppository base to maintain the same drug/solubilizer ratio. The suppositories were tested for softening time, hardness, and uniformity of mass. The dissolution test was performed using the dialysis tubing method in the basket apparatus. The amount of indomethacin released into the dissolution medium was determined spectrophotometrically at 320 nm. The results indicate that solubilization of indomethacin in the microemulsion had a positive effect on in vitro drug release but not as significant as in the case of Polysorbate 80 used alone. The enhancement ratio for Polysorbate 80 in Adeps suppositories was 2.9, for the microemulsion in Adeps suppositories was 1.1, and for Polysorbate 80 in Macrogol suppositories was 7.4 after 3 hours. The test of uniformity of mass had shown that all suppositories (reference, solubilizer-containing) are within the permitted limits. The softening time was reduced by adding the solubilizer to each type of suppository base.
直肠给药是一种合适的给药途径,对于那些对肠道非常刺激的药物(如吲哚美辛),或者当绕过肝脏的第一过效应时更有效。微乳是提高稀溶性药物渗透的一种工具。它们主要用于局部和透皮给药。然而,它们在其他给药途径中也有应用,主要是由于它们能溶解难溶性药物。栓剂碱的选择取决于药物的物理性质。本研究主要评价微乳作为稀溶性吲哚美辛在亲水性和亲脂性栓剂中的增溶剂与聚山楂酸80作为赋形剂的效果。参考栓剂采用传统的成型工艺制备,由adps solidus或Macrogol栓剂基底制备,无需药物增溶。将药物在微乳液或聚山楂酸酯80中初始增溶后,再加入栓剂碱,保持相同的药物/增溶剂比,制备微乳化栓剂。测试了栓剂的软化时间、硬度和质量均匀性。溶出度试验采用透析管法在篮式仪器中进行。采用分光光度法在320 nm处测定吲哚美辛在溶出介质中的释放量。结果表明,吲哚美辛在微乳中的增溶作用对体外药物释放有积极的影响,但不如单独使用聚山梨酯80的效果显著。3 h后,聚山梨酯80在Adeps栓剂中的增强比为2.9,微乳在Adeps栓剂中的增强比为1.1,Macrogol栓剂中的增强比为7.4。质量均匀性测试表明,所有栓剂(参比、含增溶剂)均在允许范围内。在不同类型的栓剂中加入增溶剂可以缩短软化时间。
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引用次数: 0
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European Pharmaceutical Journal
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