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Microbial canthaxanthin: an orange-red keto carotenoid with potential pharmaceutical applications. 微生物角黄素:一种具有潜在药用价值的橙红色酮类胡萝卜素。
Pub Date : 2023-09-25 eCollection Date: 2023-01-01 DOI: 10.5114/bta.2023.130733
Vinita Gaur, Surojit Bera

Canthaxanthin is an orange-red keto-carotenoid that occurs naturally and is also manufactured by synthetic methods for regular applications. In nature, canthaxanthin mainly exists in microbes such as different bacterial species, fungi, and algae, as well as in animals such as crustaceans, certain fishes, and birds. However, the amount of canthaxanthin produced in these organisms varies significantly. Additionally, the compound can be generated from genetically modified organisms using genetic engineering techniques Canthaxanthin finds extensive application as an additive in animal feed, in the pharmaceutical industry, as a coloring agent for various food products, and in cosmetics. It has powerful antioxidant properties and plays a role in lipid metabolism, neuroprotection, and immunomodulation. This article gives an extensive insight into the structure and methods of synthesis of canthaxanthin along with its various newly discovered sources identified so far. The significant applications of canthaxanthin, particularly its role in pharmaceuticals, are critically evaluated. Furthermore, the article discusses future aspects and challenges associated with canthaxanthin production and regulation.

虾青素是一种橙红色的酮类胡萝卜素,天然存在,也通过合成方法生产,用于常规应用。在自然界中,斑蝥素主要存在于微生物中,如不同的细菌、真菌和藻类,以及动物中,如甲壳类动物、某些鱼类和鸟类。然而,在这些生物体中产生的角黄素的量差异很大。此外,该化合物可以使用基因工程技术从转基因生物中产生。虾青素作为一种添加剂在动物饲料、制药工业、各种食品的着色剂和化妆品中有着广泛的应用。它具有强大的抗氧化特性,在脂质代谢、神经保护和免疫调节中发挥作用。本文对角黄素的结构、合成方法及其迄今为止新发现的各种来源进行了深入的研究。对斑蝥素的重要应用,特别是其在药物中的作用进行了批判性评价。此外,本文还讨论了斑蝥素生产和调控的未来方面和挑战。
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引用次数: 0
Impact of Enterococcus italicus ONU547 on the growth and acclimatization of micropropagated Rubus fruticosus L. and Paulownia tomentosa Steud. plants to ex vitro conditions. 意大利肠球菌ONU547对微繁殖悬钩子和毛泡桐生长和驯化的影响。植物体外条件。
Pub Date : 2023-09-25 eCollection Date: 2023-01-01 DOI: 10.5114/bta.2023.130732
Nadiia Tytarenko, Nataliia Tesliuk, Andrii Merlich, Thomas Haertlé, Volodymyr Ivanytsia

Clonal micropropagation is an effective method for plant reproduction, applicable in both scientific and industrial domains. However, a significant number of microclones are lost during the ex vitro acclimatization process. To address this, the introduction of beneficial microorganisms into the rhizosphere of micropropagated plants could have a positive effect on the survival rates and external characteristics of acclimatized plantlets. The aim of this study was to determine the protective and growth-promoting potential of Enterococcus italicus ONU547 and its effect on micropropagated plants during acclimatization. The antagonistic activity of the bacteria was determined using the agar block method. Lepidium sativum L. seeds were inoculated with bacterial suspensions at concentrations of 106, 107, and 108 CFU/ml. Subsequently, the roots of the microclones were treated with suspensions of 106 and 107 CFU/ml, and biometric characteristics were measured. The results demonstrated antagonistic properties against various phytopathogenic fungi, including Aspergillus niger, Cladosporium cladosporioides, Alternaria alternata, Alternaria tenuissima, Rhizoctonia cerealis, Penicillium expansum, and Paecilomyces variotii. Inoculation of L. sativum L. seeds resulted in improved germination rates, increased root numbers, and enhanced root and shoot lengths. Similarly, the effects of the studied bacteria on Rubus fruticosus L. and Paulownia tomentosa Steud. during the acclimatization stage led to higher survival rates, increased shoot lengths, greater node numbers, and larger leaf areas. A concentration of 107 CFU/ml was identified as optimal for inoculating the microclones. The findings indicate that E. italicus ONU547 holds promise for the inoculation of micropropagated plants during the acclimatization process. Further research is recommended to establish the specific interaction mechanisms between these bacteria and plants.

克隆微繁殖是一种有效的植物繁殖方法,适用于科学和工业领域。然而,在体外驯化过程中,大量的微克隆丢失。为了解决这一问题,将有益微生物引入微繁殖植物的根际可以对驯化植株的存活率和外部特性产生积极影响。本研究的目的是确定意大利肠球菌ONU547的保护和促生长潜力及其在驯化过程中对微繁殖植物的影响。用琼脂块法测定细菌的拮抗活性。用浓度分别为106、107和108CFU/ml的细菌悬浮液接种Lepidium sativum L.种子。随后,用106和107CFU/ml的悬浮液处理微克隆的根,并测量生物特征。结果表明,对各种植物病原真菌具有拮抗作用,包括黑曲霉、枝孢霉、交链孢、细链孢、蜡状丝核菌、扩展青霉和变异拟青霉。接种L.sativum L.种子可提高发芽率,增加根系数量,并增加根和茎的长度。同样,研究的细菌对悬钩子和毛泡桐的影响。在驯化阶段导致更高的存活率、增加的茎长、更大的节数和更大的叶面积。107CFU/ml的浓度被确定为接种微克隆的最佳浓度。研究结果表明,E.italicus ONU547有望在驯化过程中接种微繁殖植物。建议进一步研究以确定这些细菌与植物之间的特定相互作用机制。
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引用次数: 0
Physiochemical response of Cicer arietinum to zinc-containing mesoporous silica nanoparticles under water stress. 水胁迫下茜草对含锌介孔二氧化硅纳米颗粒的物理化学响应。
Pub Date : 2023-09-25 eCollection Date: 2023-01-01 DOI: 10.5114/bta.2023.130729
Maryam Mohamadzadeh, Mohsen Janmohammadi, Amin Abbasi, Naser Sabaghnia, Viorel Ion

Chickpea is an important food legume cultivated in semiarid regions, where water scarcity and nutrient deficiencies negatively affect crop production. This study aimed to investigate the effect of zinc and silicon from different sources, including bulk and nanostructures, on various biochemical traits of chickpea plants grown under field conditions in Maragheh, Northwest Iran. The main experimental factor consisted of three soil moisture levels: irrigation to 90% of field capacity (FC), 60% FC, and 30% FC. The subplots were assigned for foliar application of different fertilizers: control (distilled water), zinc sulfate (ZnSO), silicon dioxide nanoparticles (SiO2 NPs), ZnSO + SiO2 NPs, and zinc-containing mesoporous silica nanoparticles (MSNPs -Zn). The results showed that although decreased soil moisture had a negative impact on several biochemical processes, foliar application of Zn and Si in both conventional bulk and nanostructure significantly affected plant antioxidant system, plasma membrane integrity, and the concentrations of photosynthetic pigments and compatible solutes. However, the most inducing effects on catalase, ascorbate peroxidase, guaiacol peroxidase, superoxide dismutase, and anthocyanin were observed with the foliar spray of MSNPs-Zn and ZnSO + SiO2 under 60% FC. Moreover, foliar spray of MSNPs-Zn alleviated the negative effects of water deficit stress on photosynthetic pigments (chlorophyll a /b and carotenoid content). Water stress significantly induced the accumulation of free proline in the leaves. Overall, the results indicated that foliar spray of MSNPs -Zn, especially under 60% FC, improved the plant's defense system, scavenged reactive oxygen species, and enhanced the accumulation and stability of pigments, thereby mitigating the effects of drought stress.

鹰嘴豆是半干旱地区种植的重要食用豆类,缺水和营养缺乏对作物生产产生负面影响。本研究旨在研究来自不同来源的锌和硅,包括大块和纳米结构,对伊朗西北部马拉赫种植的鹰嘴豆植物在田间条件下的各种生化性状的影响。主要试验因子由三个土壤水分水平组成:灌溉至田间容量的90%、60%和30%。将亚地块分配给不同肥料的叶面施用:对照(蒸馏水)、硫酸锌(ZnSO)、二氧化硅纳米颗粒(SiO2 NPs)、ZnSO+SiO2 NPs和含锌介孔二氧化硅纳米粒子(MSNPs-Zn)。结果表明,尽管土壤水分的减少对几个生化过程产生了负面影响,但在常规本体和纳米结构中叶面施用锌和硅会显著影响植物抗氧化系统、质膜完整性以及光合色素和相容溶质的浓度。然而,在60%FC下,MSNPs-Zn和ZnSO+SiO2对过氧化氢酶、抗坏血酸过氧化物酶、愈创木酚过氧化物酶、超氧化物歧化酶和花青素的诱导作用最强。此外,叶面喷洒MSNPs-Zn减轻了水分亏缺胁迫对光合色素(叶绿素a/b和类胡萝卜素含量)的负面影响。水分胁迫显著诱导叶片中游离脯氨酸的积累。总体而言,结果表明,叶面喷施MSNPs-Zn,特别是在60%FC条件下,改善了植物的防御系统,清除了活性氧,增强了色素的积累和稳定性,从而减轻了干旱胁迫的影响。
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引用次数: 0
Expression of the gene encoding blood coagulation factor VIII without domain B in E. coli bacterial expression system. 编码无结构域B的凝血因子VIII的基因在大肠杆菌细菌表达系统中的表达。
Pub Date : 2023-09-25 eCollection Date: 2023-01-01 DOI: 10.5114/bta.2023.130728
Anna Mazurkiewicz-Pisarek, Alina Mazurkiewicz, Diana Mikiewicz, Piotr Baran, Tomasz Ciach
In this article, we have demonstrated the feasibility of generating an active form of recombinant blood coagulation factor VIII using an E. coli bacterial expression system as a potential treatment for hemophilia type A. Factor VIII (FVIII), an essential blood coagulation protein, is a key component of the fluid phase blood coagulation system. So far, all available recombinant FVIII formulations have been produced using eukaryotic expression systems. Mammalian cells can produce catalytically active proteins with all the necessary posttranslational modifications. However, cultivating such cells is time-consuming and highly expensive, and the amount of the obtained product is usually low. In contrast to eukaryotic cells, bacterial culture is inexpensive and allows the acquisition of large quantities of recombinant proteins in a short time. With this study, we aimed to obtain recombinant blood coagulation factor VIII using the E. coli bacterial expression system, a method not previously explored for this purpose. Our research encompasses the synthesis of blood coagulation factor VIII and its expression in a prokaryotic system. To achieve this, we constructed a prokaryotic expression vector containing a synthetic factor VIII gene, which was then used for the transformation of an E. coli bacterial strain. The protein expression was confirmed by mass spectrometry, and we assessed the stability of the gene construct while determining the optimal growth conditions. The production of blood coagulation factor VIII by the E. coli bacterial strain was carried out on a quarter-technical scale. We established the conditions for isolation, denaturation, and renaturation of the protein, and subsequently confirmed the activity of FVIII.
在这篇文章中,我们已经证明了使用大肠杆菌细菌表达系统产生活性形式的重组凝血因子VIII作为a型血友病的潜在治疗方法的可行性。因子VIII(FVIII)是一种必需的凝血蛋白,是液相凝血系统的关键组分。到目前为止,所有可用的重组FVIII制剂都是使用真核表达系统生产的。哺乳动物细胞可以产生具有所有必要翻译后修饰的催化活性蛋白质。然而,培养这样的细胞是耗时且高成本的,并且所获得的产物的量通常较低。与真核细胞相比,细菌培养成本低廉,可以在短时间内获得大量重组蛋白。在这项研究中,我们旨在使用大肠杆菌表达系统获得重组凝血因子VIII,这是一种以前没有探索过的方法。我们的研究包括凝血因子VIII的合成及其在原核系统中的表达。为了实现这一点,我们构建了含有合成因子VIII基因的原核表达载体,然后将其用于转化大肠杆菌菌株。蛋白质表达通过质谱法得到证实,我们在确定最佳生长条件的同时评估了基因构建体的稳定性。由大肠杆菌菌株生产凝血因子VIII是在四分之一的技术规模上进行的。我们建立了分离、变性和复性蛋白质的条件,随后证实了FVIII的活性。
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引用次数: 0
Effect of sucrose as an elicitor in increasing quercetin-3-O-rhamnoside (quercitrin) content of chrysanthemum (Chrysanthemum morifolium Ramat) callus culture based on harvest time differences. 基于收获时间差异,蔗糖作为诱导子在提高菊花愈伤组织培养中槲皮素-3-O-鼠李糖苷(槲皮素)含量方面的作用。
Pub Date : 2023-09-25 eCollection Date: 2023-01-01 DOI: 10.5114/bta.2023.130731
Tia Setiawati, Annisa N Arofah, Mohamad Nurzaman, Annisa Annisa, Asep Z Mutaqin, Rusdi Hasan

Chrysanthemum (Chrysanthemum morifolium) contains secondary metabolites, such as flavonoid compounds, especially luteolin-7-glucoside and quercetin-3-O-rhamnoside (quercitrin), in its tissues. Utilizing sucrose as an elicitor through callus culture presents an alternative method to enhance the production of secondary metabolites. This research aimed to determine the best sucrose concentration and harvest time for maximizing quercitrin content in chrysanthemum callus culture. The research employed a completely randomized design with four treatment groups: 0, 30, 45, and 60 g/l of sucrose added to MS medium containing 4 ppm 2,4-dichlorophenoxyacetic acid (2,4-D). Callus samples were harvested on the 15th and 30th days of culture. The observed parameters included callus morphology (color and texture), fresh weight, dry weight, the diameter of the callus, and quercitrin content analyzed using high-performance liquid chromatography. The results showed that all callus cultures exhibited intermediate textures and varied colors, predominantly shades of brown. The treatment involving 45 g/l of sucrose with a 30th-day harvest yielded the highest fresh weight, dry weight, and quercitrin content, namely 2.108 g, 0.051 g, and 0.437 mg/g DW, respectively. Notably, the quercitrin content exhibited a 63.67% increase compared to the control.

菊花(菊花)在其组织中含有次生代谢产物,如类黄酮化合物,特别是木犀草素-7-葡萄糖苷和槲皮素-3-O-鼠李糖苷(槲皮素)。利用蔗糖作为诱导子通过愈伤组织培养提供了一种提高次生代谢产物产生的替代方法。本研究旨在确定菊花愈伤组织培养中最大限度提高槲皮素含量的最佳蔗糖浓度和收获时间。该研究采用了一种完全随机的设计,分为四个处理组:0、30、45和60克/升蔗糖添加到含有4 ppm 2,4-二氯苯氧乙酸(2,4-D)的MS培养基中。在培养的第15天和第30天收获愈伤组织样品。观察到的参数包括愈伤组织的形态(颜色和质地)、鲜重、干重、愈伤组织的直径以及用高效液相色谱分析的槲皮素含量。结果表明,所有愈伤组织培养物都表现出中等质地和不同的颜色,主要是棕色。使用45g/l蔗糖和第30天收获的处理产生最高的鲜重、干重和槲皮素含量,分别为2.108g、0.051g和0.437mg/g DW。值得注意的是,与对照相比,槲皮素含量增加了63.67%。
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引用次数: 0
Identification of potential CD8+ epitopes in pp62 polyprotein of African swine fever virus using computational immunology. 使用计算免疫学鉴定非洲猪瘟病毒pp62多蛋白中潜在的CD8+表位。
Pub Date : 2023-09-25 eCollection Date: 2023-01-01 DOI: 10.5114/bta.2023.130726
Mark Lester C Galicia, Dale Jonathan M Morales, Precious Grace B Pogado, Ashley L Quebrado, Leana Rich Herrera-Ong

The highly infectious African swine fever virus (ASFV) is currently the only known DNA arbovirus within the Asfarviridae family which primarily infects domestic pigs and wild boars. African swine fever (ASF) leads to a mortality rate of up to 100% which has caused massive socio-economic losses worldwide. Previous research indicates that ASFV's virulence can be attributed to polyprotein pp62, which plays a crucial role in viral assembly and core maturation. This particular study utilized in silico analysis to identify highly conserved cytotoxic T-cell epitopes in pp62 that can potentially serve as key components for future ASFV vaccines. To achieve this, the researchers retrieved, clustered, and aligned the peptide sequences of pp62. Subsequently, the aligned sequences were analyzed to identify epitopes that bind promiscuously to the swine major histocompatibility complex I (MHC I) alleles and exhibiting MHC IC50 values < 500 nM. Additionally, peptide sequences with positive proteasome and TAP scores were considered. Potential cross-reactivity was assessed by comparing the peptide sequences against available proteome sequences of Sus scrofa domesticus in various databases. Furthermore, molecular docking was conducted to evaluate the binding of candidate epitopes with swine leukocyte antigen-1*0401 (SLA-1*0401). The dissociation constants, binding energies, root mean square deviation, and root mean square fluctuation values for the SLA-epitope complexes were compared with a positive reference. In the course of the study, 21 highly conserved CD8+ epitopes were identified, out of which four were further assessed for their potential immunogenicity. The results demonstrated that the highly conserved CD8+ epitopes discovered in this study are promising for integration into future ASFV vaccine formulations. As preliminary data, it is anticipated that these findings will subsequently undergo in vitro and in vivo studies in the future.

传染性极强的非洲猪瘟病毒(ASFV)是目前已知的唯一一种主要感染家猪和野猪的阿氏病毒科DNA虫媒病毒。非洲猪瘟的死亡率高达100%,在全球范围内造成了巨大的社会经济损失。先前的研究表明,ASFV的毒力可归因于多蛋白pp62,它在病毒组装和核心成熟中起着至关重要的作用。这项特殊的研究利用计算机分析来鉴定pp62中高度保守的细胞毒性T细胞表位,这些表位可能成为未来ASFV疫苗的关键成分。为了实现这一点,研究人员对pp62的肽序列进行了检索、聚类和比对。随后,对比对的序列进行分析,以鉴定与猪主要组织相容性复合体I(MHC I)等位基因混杂结合并显示MHC IC50值<500nM的表位。此外,还考虑了蛋白酶体和TAP评分为阳性的肽序列。通过将肽序列与各种数据库中Sus scrofa domesticus的可用蛋白质组序列进行比较来评估潜在的交叉反应性。此外,进行分子对接以评估候选表位与猪白细胞抗原-1*0401(SLA-1*0401)的结合。将SLA表位复合物的解离常数、结合能、均方根偏差和均方根波动值与阳性参考进行比较。在研究过程中,鉴定了21个高度保守的CD8+表位,其中4个表位的潜在免疫原性得到了进一步评估。结果表明,本研究中发现的高度保守的CD8+表位有望整合到未来的ASFV疫苗配方中。作为初步数据,预计这些发现将在未来进行体外和体内研究。
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Biotechnologia
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