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Cytotoxic of Usnic Acid Isolated from Ramalina sp. 从 Ramalina sp.
Q4 Agricultural and Biological Sciences Pub Date : 2024-07-17 DOI: 10.11594/jtls.14.02.14
Akhmad Darmawan, M. Maulidiyah, Megawati Megawati, Novita Ariani, Sitti Aisya, S. Sukirno, Ahmad Randy, G. Primahana, Medi Hendra, Muhammad Nurdin
Ramalina sp. (Ramalinaceae) is a type of lichen known to contain many active secondary metabolite compounds that have the potential to be used as medicine or medicinal raw materials. One of the biological activities possessed by Ramalina sp. lichen is its anticancer activity. This research aims to isolate and identify active secondary metabolite compounds from the methanol extract of the Ramalina sp. lichen and to find out the cytotoxic activity of the isolated compound against MCF7 breast cancer cells. Compound 1 (usnic acid) was successfully isolated from fraction A. The isolation and purification process was carried out starting with a maceration process using acetone solvent, followed by silica gel column chromatography using a gradient solvent system consisting of n-hexane, n-hexane: EtOAc, EtOAc, EtOAc: MeOH, and MeOH with 5% increment of polarity to obtain 17 fractions (F-1 to F-17). From the 17 fractions obtained, fraction 3 (F-3) and fraction 4 (F-4) (eluted with n-hexane: EtOAc 30%), which had the same TLC profile, were combined and named as fraction A. Compound 1 (50 mg) is a yellow needle crystal that was formed in a bottle of fraction A, which was obtained after the process of combining fractions F-3 and F-4 and solvent evaporation process. The crystals were then separated and purified with CHCl3 and MeOH. Compound 1 was then characterized based on spectroscopic data. Various spectroscopic analysis data, including FTIR, 1D- and 2D-NMR, and LC-ESI-MS, show that Compound 1 is a dibenzofuran derivative compound with 18 carbons (3 from carbonyl groups (C=O) and 3 from methyl groups) and 2 hydroxyls (-OH). Cytotoxicity assay showed that at a low concentration of 18.75 ug/mL, Com-pound 1 caused a 67.06% decrease in MCF7 viability.
Ramalina sp.(Ramalinaceae)是一种地衣,已知其含有许多活性次生代谢化合物,具有作为药物或药用原料的潜力。Ramalina 地衣所具有的生物活性之一是其抗癌活性。本研究旨在从 Ramalina sp. 地衣的甲醇提取物中分离和鉴定活性次生代谢物化合物,并研究分离出的化合物对 MCF7 乳腺癌细胞的细胞毒活性。化合物 1(usnic acid)从馏分 A 中成功分离出来。分离和纯化过程首先使用丙酮溶剂进行浸渍,然后使用硅胶柱色谱法,其梯度溶剂系统包括正己烷、正己烷、EtOAc、EtOAc、EtOAc、EtOAc、EtOAc、EtOAc、EtOAc、EtOAc、EtOAc、EtOAc:EtOAc、EtOAc、EtOAc:MeOH,以及极性递增 5% 的 MeOH,得到 17 个馏分(F-1 至 F-17)。从得到的 17 份馏分中,将具有相同 TLC 图谱的馏分 3(F-3)和馏分 4(F-4)(用正己烷:EtOAc 30%洗脱)合并,命名为馏分 A。晶体经分离后用 CHCl3 和 MeOH 纯化。然后根据光谱数据对化合物 1 进行表征。各种光谱分析数据,包括傅立叶变换红外光谱、一维和二维核磁共振以及 LC-ESI-MS 显示,化合物 1 是一种二苯并呋喃衍生物化合物,含有 18 个碳原子(3 个来自羰基(C=O),3 个来自甲基)和 2 个羟基(-OH)。细胞毒性试验表明,在 18.75 微克/毫升的低浓度下,化合物 1 可使 MCF7 的存活率降低 67.06%。
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引用次数: 0
Mycobacterium leprae Survival Inside Acanthamoeba sp. Isolated from Water Source in Leprosy Endemic Area, Indonesia 麻风分枝杆菌在印度尼西亚麻风病流行区水源分离出的棘阿米巴中的存活率
Q4 Agricultural and Biological Sciences Pub Date : 2024-07-17 DOI: 10.11594/jtls.14.02.07
Ratna Wahyuni, Linda Astari, I. Iswahyudi, Sepling Paling, D. Adriaty, Siti Kurniawati, Syifa Aulia, Bandaru Rahmatari, C. Prakoeswa, I. Agusni, Shinzo Izumi
Mycobacterium leprae is an acid-fast bacterium that causes leprosy diseases, which remains a problem worldwide. Even though leprosy prevalence in the world has decreased significantly, many endemic pocket areas continue reporting new cases and harbor M. leprae in the environment, including water and soil. The presence of obligate intracellular bacteria-M. leprae in the environment raises a question on how it survives. Free-living amoeba has been proposed as its reservoir host in the environment. The study was conducted to give evidence that M. leprae can survive inside free-living amoeba isolated from water sources of leprosy endemic areas. M. leprae from leprosy patients was cultured together with Acanthamoeba sp. isolated from the water source of the leprosy endemic area. Viability and duplication of M. leprae inside amoeba then observed at days 14 and 28 using reverse transcriptase PCR and qPCR. The results showed that M. leprae survived inside the amoeba until day 28, but no bacterial replication was observed. The study reveals in vitro evidence of viable M. leprae inside free-living amoeba of leprosy endemic area environment
麻风分枝杆菌是一种导致麻风病的酸性无菌细菌,麻风病仍然是一个世界性问题。尽管麻风病在全球的流行率已大幅下降,但许多麻风病流行的袖珍地区仍不断报告新病例,并在水和土壤等环境中孳生麻风分枝杆菌。环境中存在麻风杆菌这种必须存在于细胞内的细菌,这就提出了麻风杆菌如何生存的问题。有人提出,自由生活的变形虫是其在环境中的贮存宿主。这项研究旨在证明麻风杆菌可以在从麻风病流行地区的水源中分离出来的自由生活阿米巴中存活。研究人员将麻风病人体内的麻风杆菌与从麻风病流行区水源中分离出的棘阿米巴原虫一起进行培养。然后利用逆转录酶 PCR 和 qPCR 技术,在第 14 天和第 28 天观察麻风杆菌在阿米巴体内的存活率和复制情况。结果显示,麻风杆菌在变形虫体内存活到第 28 天,但没有观察到细菌复制。该研究揭示了麻风病流行地区环境中自由生活的变形虫体内存在可存活的麻风杆菌的体外证据。
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引用次数: 0
Effect of Multiple Types of High-fat Diet Induction on Gut Microbiota Profile and Butyrate Levels on NAFLD Rat Model 多种高脂饮食诱导对非酒精性脂肪肝模型大鼠肠道微生物群谱和丁酸盐水平的影响
Q4 Agricultural and Biological Sciences Pub Date : 2024-07-17 DOI: 10.11594/jtls.14.02.08
Syifa Mustika, Nabila Ramadhani, Novita Apramadha Kartika Sari, Dian Handayani, Dewi Santosaningsih
Dysbiosis, marked by a decline in Firmicutes (such as Clostridium spp.) and a rise in Proteobacteria (such as Escherichia coli), is recognized as a pathway linked to the progression of non-alcoholic fatty liver disease (NAFLD). High-fat food may influence the development of NAFLD. This study investigates the effect of different high-fat diets on gut microbiota composition and butyrate levels in a rat model. Four groups of Rattus norvegicus strain Wistar (n=27) were fed specific diets for 12 weeks: normal diet (ND, n=7), high-fat diet (HFD, n=6), western diet (WD, n=7), and high-fat-high-fructose diet (HFHFD, n=7). Stools were cultured on Eosin Methylene Blue (EMB) for Escherichia coli and Forget Fredette Agar (FFA) for Clostridium spp. Colonies were counted using Total Plate Count (TPC), and butyrate levels were measured with gas chromatography. Liver histology was assessed with the NAFLD activity score (NAS). Gut microbiota were analyzed using Crosstab and Chi-Square tests, and butyrate levels with the Kruskal-Wallis test. Butyrate levels decreased in NAFLD-inducing diet groups (HFD, WD, HFHFD) compared to ND (p=0.021). Escherichia coli colony counts were significantly higher in NAFLD-inducing diet groups (p=0.048), while Clostridium spp. counts were higher in ND (p=0.001). Liver histological changes, particularly in the WD group, resembled NAFLD. These findings imply that different high-fat diets can alter the gut microbiome, potentially influencing NAFLD development.
菌群失调的标志是固醇菌(如梭状芽孢杆菌属)的减少和蛋白菌(如大肠杆菌)的增加,被认为是与非酒精性脂肪肝(NAFLD)的发展有关的途径。高脂肪食物可能会影响非酒精性脂肪肝的发展。本研究调查了不同高脂饮食对大鼠肠道微生物群组成和丁酸水平的影响。对四组野鼠品系 Wistar(n=27)喂食特定饮食 12 周:正常饮食(ND,n=7)、高脂饮食(HFD,n=6)、西式饮食(WD,n=7)和高脂-高果糖饮食(HFHFD,n=7)。粪便经甲亚砜蓝(EMB)培养以检测大肠埃希氏菌,忘弗雷德特琼脂(FFA)培养以检测梭状芽孢杆菌。菌落计数采用总平板计数法(TPC),丁酸盐含量采用气相色谱法测量。肝脏组织学用非酒精性脂肪肝活动评分(NAS)进行评估。肠道微生物群采用交叉分析和奇偶检验进行分析,丁酸盐水平采用 Kruskal-Wallis 检验进行分析。与ND相比,NAFLD诱导饮食组(HFD、WD、HFHFD)的丁酸盐含量有所下降(p=0.021)。非酒精性脂肪肝诱导饮食组的大肠埃希氏菌落计数明显较高(p=0.048),而玖玖组的梭状芽孢杆菌计数较高(p=0.001)。肝脏组织学变化,尤其是在 WD 组,与非酒精性脂肪肝相似。这些研究结果表明,不同的高脂饮食会改变肠道微生物组,从而可能影响非酒精性脂肪肝的发生。
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引用次数: 0
Validation of Hand Grading Technique of Corpora Lutea in Embryo Recipient Cows 胚胎受体奶牛黄体手工分级技术的验证
Q4 Agricultural and Biological Sciences Pub Date : 2024-07-17 DOI: 10.11594/jtls.14.02.09
Gamage Dammika Rohan Kumara Perera, Pupulewatte Gedara Anil Pushpakumara, Ponweera Arachchige Basil Douglas Alexander
The success of embryo transfer in cattle may depend on several factors. The blood progesterone level of the embryo recipient would be one of the main factors secondary to the size and quality of the corpus luteum. Per-rectal palpation and by-hand grading of corpora lutea have been practiced routinely, even under commercial embryo transfer programs in cattle. This experiment aimed to evaluate the accuracy of the per-rectal hand grading technique of mid-phase corpora lutea in embryo recipient cows. A group of healthy cows (Friesian× Jersey, n=25), which had been selected as recipient animals with the special consideration of reproductive history in an embryo transfer program, was used in this study. Estrous of those animals were synchronized using a standard progesterone intravaginal devise-PGF2α protocol. Conven-tional evaluation and grading of corpora lutea were conducted at the per-rectal hand grading as 'A', 'B', 'C' and 'Cystic' (If diameters; < 1cm=C, 1-1.9cm=B, 2-2.5cm= A, >2.5cm= Cystic). In this study, the accuracy of those hand-grading techniques was evaluated using ultra-sound scanning technology on the same day. Identification accuracy of grades C, B, A, and Cysts in hand grading technique was 45%, 56%, 67%, and 50%, respectively. The average accuracy in the hand grading technique was 52%, and identification accuracy as suitable (A or B) corpora lutea for embryo transfer was 76%. Results raised the 24% probable loss of embryo transferrable recipients due to the hand-grading technique. Further, 8% of unsuitable cows can be selected as embryo recipients due to the practice of hand-grading techniques at embryo transfer programs. Although higher accuracy is a characteristic of the hand grading technique, the technology causes considerable loss due to inaccurate selection probability of recipients at embryo transfer programs.
牛的胚胎移植成功与否取决于多个因素。胚胎受体的血液孕酮水平是主要因素之一,其次是黄体的大小和质量。即使在牛的商业胚胎移植项目中,直肠触诊和人工黄体分级也是常规做法。本实验旨在评估胚胎受体奶牛直肠周围黄体中期手工分级技术的准确性。本研究使用了一组健康的奶牛(弗里斯兰牛×娟珊牛,n=25),这些奶牛被选为胚胎移植项目中的受体动物,并特别考虑了其繁殖史。使用标准的阴道内黄体酮装置-PGF2α方案对这些动物的雌性期进行同步。在直肠周围对黄体进行常规评估和分级,分为 "A"、"B"、"C "和 "囊肿 "四级(直径小于 1 厘米=C,1-1.9 厘米=B,2-2.5 厘米=A,大于 2.5 厘米=囊肿)。在本研究中,使用超声波扫描技术在同一天对这些手工分级技术的准确性进行了评估。用手工分级技术鉴别 C、B、A 和囊肿的准确率分别为 45%、56%、67% 和 50%。手工分级技术的平均准确率为 52%,而鉴定为适合胚胎移植的(A 或 B)黄体的准确率为 76%。结果表明,由于手工分级技术,胚胎移植受体可能损失 24%。此外,由于胚胎移植项目采用手工分级技术,8%的不合适奶牛可能被选为胚胎受体。虽然精确度较高是手工分级技术的一个特点,但由于胚胎移植项目选择受体的概率不准确,该技术造成了相当大的损失。
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引用次数: 0
The Role of Mycorrhizae on the Growth of Banana Planlets of Cultivar Raja (Musa Paradisiaca C.V. Raja) from Bali at Post Acclimatization 菌根对巴厘岛栽培品种拉贾(Musa Paradisiaca C.V. Raja)小香蕉在适应后生长的作用
Q4 Agricultural and Biological Sciences Pub Date : 2024-07-17 DOI: 10.11594/jtls.14.02.11
R. Dwiyani, I. Wirya, I. G. A. Gunadi, I. A. P. Darmawati, Hestin Yuswanti, Ketut Budi Susrusa, A. A. M. Astiningsih
The effect of inoculation of a combination of the arbuscular mycorrhizal fungus of Glomus and Acaulospora on micropropagated banana plantlets cultivar Raja from Bali during the acclimatization phase was investigated. The experiment was conducted at the Experimental Station of the Faculty of Agriculture, Udayana University, Denpasar, Bali, Indonesia, over the period of March to June 2022. The objective of this research was to evaluate the impact of various doses of mycorrhizae on the growth of tissue culture banana plantlets that had undergone acclimatization for a period of six weeks. A total of five different doses of mycorrhiza were applied to the acclimatized plantlets, namely M0=without mycorrhiza and M5=5g mycorrhizal. kg1, M10=10 g mycorrhiza.kg-1, M15=15 g mycorrhiza.kg-1, M20=20 g mycorrhizal. kg 1. A significant effect was discovered at M15; however, compared to M20, M20 gave better results for root length and fresh weight of the above-ground part, indicating the best dose was 20 g mycorrhiza kg-1. This study concluded that mycorrhiza has a very significant role in supporting the growth of micro-propagated banana plantlets at post-acclimatization. Variables observed, such as the average number of leaves, stem diameter, above-ground plant fresh and dry weight, and root fresh and dry weight, increased with the application of AMF (Glomus and Acaulospora).
研究了在适应阶段接种 Glomus 和 Acaulospora 的丛枝菌根真菌组合对巴厘岛 Raja 栽培品种香蕉小苗的影响。实验于 2022 年 3 月至 6 月在印度尼西亚巴厘岛登巴萨市乌达亚纳大学农学院实验站进行。这项研究的目的是评估不同剂量的菌根对经过六周适应期的香蕉组培苗生长的影响。研究人员对经过驯化的香蕉小苗施用了五种不同剂量的菌根,即 M0= 无菌根和 M5=5g 菌根.kg1、M10=10g 菌根.kg-1、M15=15g 菌根.kg-1、M20=20g 菌根.kg1。发现 M15 有明显效果;但与 M20 相比,M20 在根长和地上部分鲜重方面效果更好,表明最佳剂量为 20 克菌根 kg-1。这项研究的结论是,菌根对微繁香蕉植株在后气候条件下的生长具有非常重要的支持作用。所观察到的变量,如平均叶片数、茎直径、地上部植物鲜重和干重、根鲜重和干重,都随着施用 AMF(Glomus 和 Acaulospora)而增加。
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引用次数: 0
Antioxidant Activity of Baby Java Citrus Peel Extract Promotes Lung Tissue Repair in Mice Challenged by Lipopolysaccharides 爪哇宝宝柑橘皮提取物的抗氧化活性可促进受到脂多糖挑战的小鼠肺组织修复
Q4 Agricultural and Biological Sciences Pub Date : 2024-07-17 DOI: 10.11594/jtls.14.02.03
Farida Rachmawati, F. Sari, Sefihara Paramitha Sakti, Muhammad Wisam Wira Sakti, Sri Rahayu, A. Soewondo, M. Rifa’i
Acute lung injury tends to be induced by infection or sepsis that disrupts alveolar and vascular permeability, neutrophil influx, and edema. Those impairments are worsened by the increase of oxidative stress along with hyperinflammation response. Oxidative stress in lung tissue could be indicated by malondialdehyde (MDA) and nuclear fac-tor erythroid 2-related factor 2 (Nrf2) expression. This research aimed to evaluate the efficacy of Baby Java citrus peel extract (BJE) in suppressing oxidative stress and preventing lung injury in lipopolysaccharides (LPS)-induced mice. Twenty-five male BALB/c mice were divided into five groups consisting of untreated (N), LPS (A), and LPS-induced followed by treatment using BJE at various doses: 75 mg/kg BW (BJE-1), 105 mg/kg BW (BJE-2), and 150 mg/kg BW (BJE-3). Lungs were isolated for histopathological analysis also detection of MDA and Nrf2 using flow cytometry. BJE at the dose of 105 mg/kg BW could inhibit the alteration of lung histology following LPS challenge including alveolar and interstitial neutrophil infiltration, proteinaceous debris, and septal thickening. The same dose also showed good potency in suppressing MDA and Nrf2 levels as oxidative stress indicators. Our findings demonstrated the protective effects of Baby Java citrus peel in acute lung injury and oxidative stress prevention after LPS exposure
感染或败血症往往会诱发急性肺损伤,破坏肺泡和血管的通透性、中性粒细胞涌入和水肿。氧化应激和高炎症反应的增加会加重这些损伤。肺组织中的氧化应激可通过丙二醛(MDA)和核红细胞2相关因子2(Nrf2)的表达来反映。本研究旨在评估爪哇宝宝柑橘皮提取物(BJE)在抑制氧化应激和预防脂多糖(LPS)诱导的小鼠肺损伤方面的功效。25 只雄性 BALB/c 小鼠被分为五组,包括未处理组(N)、LPS(A)和 LPS 诱导组,然后使用不同剂量的 BJE 进行处理:75毫克/千克体重(BJE-1)、105毫克/千克体重(BJE-2)和150毫克/千克体重(BJE-3)。分离肺脏进行组织病理学分析,并使用流式细胞术检测 MDA 和 Nrf2。剂量为 105 毫克/千克体重的 BJE 可抑制 LPS 挑战后肺组织学的改变,包括肺泡和间质中性粒细胞浸润、蛋白碎屑和隔膜增厚。同样的剂量还能有效抑制作为氧化应激指标的 MDA 和 Nrf2 水平。我们的研究结果表明,爪哇宝宝柑橘皮对急性肺损伤和 LPS 暴露后氧化应激的预防具有保护作用。
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引用次数: 0
An Overview of Traditional Medicinal Plants Used in Treating Hepatocellular Carcinoma (HCC) with Emphasis on Mechanisms of Action 用于治疗肝细胞癌(HCC)的传统药用植物概述,重点介绍其作用机制
Q4 Agricultural and Biological Sciences Pub Date : 2024-07-17 DOI: 10.11594/jtls.14.02.20
Habeebulahi Ajibola Adekilekun
Cancer is an incurable fatal disease wreaking havoc on the population of countries either developed or developing. Hepatocellular carcinoma (HCC) is the fifth most frequent type of cancer having a poor prognosis. Globally, HCC management and treatment intervention using medicinal plants from generation to generation is rapidly increasing and remains an area of promising research. This review provided insight into medicinal plants' ethnomedicinal use, photochemistry and mechanisms of action on HCC exhibiting anti-cancer potentials subjected to scientific investigations. Furthermore, clinical, toxicity studies and detailed mechanisms of action would provide an understanding of the development of lead compounds in drug discovery and development for HCC.
癌症是一种无法治愈的致命疾病,对发达国家或发展中国家的人口造成严重破坏。肝细胞癌(HCC)是第五大常见癌症类型,预后极差。在全球范围内,一代又一代人利用药用植物进行 HCC 管理和治疗干预的情况正在迅速增加,这仍然是一个前景广阔的研究领域。本综述深入探讨了药用植物的民族药用用途、光化学作用以及对 HCC 的作用机制,并对其抗癌潜力进行了科学研究。此外,临床、毒性研究和详细的作用机制将有助于了解 HCC 药物研发中先导化合物的开发情况。
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引用次数: 0
Different Mangrove Rehabilitation Statuses Effects to Benthic Bacterial Structure Community in the Northern Area of Java Island 不同红树林恢复状况对爪哇岛北部地区底栖细菌结构群落的影响
Q4 Agricultural and Biological Sciences Pub Date : 2024-07-17 DOI: 10.11594/jtls.14.02.15
V. Vidayanti, C. Retnaningdyah, Irfan Mustafa, Indah Oktaviani
The study proposed identifying the changes in bacterial community and diversity and exploring the potential correlations among sediment parameters and benthic bacterial communities under different mangrove ecosystem rehabilitation statuses. Three sites were investigated, 1. Kampung Blekok (KB) as the long rehabilitation period site; 2. Banyuurip Mangrove Center (BMC) as the short rehabilitation period site; 3. Pulau Lusi (PL) as the reclamation site. The physicochemical parameters and benthic bacteria from the mangrove sediment of the three locations were observed. The data were analyzed statistically to determine the sites' variation and the correlation between parameters. The results found that the physicochemical parameters among restoration sites varied. Soil organic matter (SOM) in long and short-rehabilitation period sites was more than 1.5-fold higher than those in the reclamation site. Cation exchange capacity (CEC), calcium and potassium ions, and soil conductivity of the restoration site have higher values and can reach more than 1.3 fold compared to the other sites. According to diversity indices and taxa richness, benthic bacteria in the restoration were the most diverse. Proteobacteria dominated in natural and restoration sites; meanwhile, Firmicutes dominated in the reclamation site. Sulfurovum aggregans were found abundant in the long and short rehabilitation sites; meanwhile, Mesobacillus subterraneus was the dominant species in the reclamation site. Furthermore, the bacterial taxa richness was positively correlated with SOM and the bacterial diversity was correlated with CEC and conductivity.
该研究建议确定细菌群落和多样性的变化,并探索不同红树林生态系统恢复状态下沉积物参数与底栖细菌群落之间的潜在相关性。研究调查了三个地点:1.甘榜布勒谷(KB),作为长期恢复地点;2.班佑里红树林中心(BMC),作为短期恢复地点;3.卢西岛(PL),作为填海地点。对三个地点的红树林沉积物的物理化学参数和底栖细菌进行了观察。对数据进行了统计分析,以确定三个地点的差异和参数之间的相关性。结果发现,不同修复地点的理化参数存在差异。长恢复期和短恢复期地点的土壤有机质(SOM)是填海地点的 1.5 倍以上。恢复点的阳离子交换容量(CEC)、钙离子、钾离子和土壤电导率值较高,可达其他恢复点的 1.3 倍以上。从多样性指数和类群丰富度来看,修复区的底栖细菌种类最多。自然区和修复区的底栖细菌以蛋白质细菌为主,而填海区则以固着菌为主。Sulfurovum aggregans(聚合硫杆菌)在长短期修复地点均有大量发现;而在填海地点,Mesobacillus subterraneus 是优势菌种。此外,细菌类群丰富度与 SOM 呈正相关,细菌多样性与 CEC 和电导率呈正相关。
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引用次数: 0
Three-Dimensional Structure of Human Epididymis Protein 4 (HE4): A Protein Modelling of an Ovarian Cancer Biomarker Through In Silico Approach 人类附睾蛋白 4 (HE4) 的三维结构:通过硅学方法建立卵巢癌生物标志物的蛋白质模型
Q4 Agricultural and Biological Sciences Pub Date : 2024-07-17 DOI: 10.11594/jtls.14.02.13
Nur Nadiah Abdul Rashid, Mohd Hamzah Mohd Nasir, Nurasyikin Hamzah, Che Muhammad Khairul Hisyam Ismail, Siti Aishah Sufira Nor Hishamuddin, I. Suffian, Azzmer Azzar Abdul Hamid
The Human Epididymis Protein 4 (HE4) biomarker has been extensively investigated for its potential in diagnosing ovarian cancer (OC). For the application of diagnostic techniques and drug delivery, it is crucial to understand the protein tertiary structure. However, the Protein Data Bank (PDB) does not currently contain the three-dimensional (3D) structure of HE4. Therefore, an in silico analysis was conducted to model the HE4 protein using AlphaFold, I-TASSER, and Robetta servers, with the sequence retrieved from UniProt (ID: Q14508). These three servers employed deep learning algorithms, threading templates, and de novo methods, respectively. Subsequently, Molecular Dynamics (MD) simulation using the GROMACS software package improved each 3D structure model, resulting in optimized and refined structures: RF1, RF2, and RF3. PROCHECK and ERRAT programs were employed to assess the structure quality. The Ramachandran plots from PROCHECK indicated that 100% of residues were within the allowed regions for all servers except for I-TASSER. For the refined structures, RF1 and RF3, all residues were concentrated within the allowed regions. According to the ERRAT program, the RF1 model exhibited the highest overall quality factor of 97.701, followed by RF3 and AlphaFold models with scores of 94.643 and 93.750, respectively. After these validations, RF1 emerged as the most accurately predicted 3D structure of HE4 and has one tunnel identified by CAVER 3.0 tool that facilitates the transportation of small particles to the active site, supported by FTsite and PrankWeb binding site predictions. This model holds potential for various computational studies, including the development of OC diagnostic kits. It will enhance our comprehension of the interactions between the protein and other biomolecules.
人类附睾蛋白 4(HE4)生物标志物在诊断卵巢癌(OC)方面的潜力已得到广泛研究。要应用诊断技术和给药技术,了解蛋白质的三级结构至关重要。然而,蛋白质数据库(PDB)目前并不包含 HE4 的三维(3D)结构。因此,我们使用 AlphaFold、I-TASSER 和 Robetta 服务器,利用从 UniProt(ID:Q14508)检索到的序列,对 HE4 蛋白质进行了硅学分析建模。这三个服务器分别采用了深度学习算法、穿线模板和全新方法。随后,使用 GROMACS 软件包进行分子动力学(MD)模拟,改进了每个三维结构模型,从而得到了优化和完善的结构:RF1、RF2 和 RF3。PROCHECK 和 ERRAT 程序用于评估结构质量。PROCHECK 的拉马钱德兰图显示,除 I-TASSER 外,所有服务器中 100%的残基都在允许区域内。对于 RF1 和 RF3 的精炼结构,所有残基都集中在允许区域内。根据ERRAT程序,RF1模型的总体质量因子最高,为97.701,其次是RF3和AlphaFold模型,分别为94.643和93.750分。经过这些验证后,RF1 成为预测最准确的 HE4 三维结构,它有一个由 CAVER 3.0 工具确定的隧道,有助于将小颗粒运送到活性位点,并得到了 FTsite 和 PrankWeb 结合位点预测的支持。该模型可用于各种计算研究,包括开发 OC 诊断试剂盒。它将增强我们对蛋白质与其他生物大分子之间相互作用的理解。
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引用次数: 0
Increase in Serum Ferritin Level as a Marker of Disease Activity in Pediatric Sys-temic Lupus Erythematosus (pSLE) Patients 血清铁蛋白水平升高作为儿童系统性红斑狼疮(pSLE)患者疾病活动的标志
Q4 Agricultural and Biological Sciences Pub Date : 2023-05-25 DOI: 10.11594/jtls.13.02.11
D. Wulandari, W. Barlianto, Susanto Nugroho, T. Sari
Ferritin is an acute-phase reactant that is elevated in autoimmune disorders, including systemic lupus erythematosus (SLE). However, their correlation with disease activity scores has not been confirmed. Pandemic Covid-19 makes children visitation to hospital to get the treatment of SLE were delayed. This study aimed to evaluate correlation between serum ferritin and disease activity and its role in screening for flare in pediatric SLE (pSLE) patients during pandemic Covid-19. This is a cross-sectional study conducted in Saiful Anwar General Hospital Malang. Sampling was carried out sequentially on pediatric patients who met the criteria for Systemic Lupus International Collaborating Clinics (SLICC) and were recorded between July 2021-May 2022. All patients were interviewed and assessed for disease activity using SLE Disease Activity Index 2000 (SLEDAI-2k). A score <4 was categorized as inactive disease. Biochemical, serological tests including markers of disease activity and ferritin level were measured by standard laboratory procedure. Comparison, correlation and ROC curve analyses were performed with SPSS software. There were 38 females pSLE participated in this study. The mean age of the patients were 12.6 ± 3.02 years. Serum ferritin significantly higher in active disease compared to inactive disease 84.50 ng/mL (68.00-151.75 ng/mL) ng/mL and 815.00 ng/mL (451.25-1570.00 ng/mL), a value of p<0.05 was determined to be statistically significant. A significant correlation was found between serum ferritin with SLEDAI 2K (r = 0.890, p = 0.000). Correlation was also found between serum ferritin and IgM anti-double stranded-DNA (r = 0.325, p = 0.046), but not with other laboratory and serological parameters. In ROC curve analysis, we found that Area Under The Curve (AUC) 0.989, 95%CI 0.964-1.014, p value 0,000, with cut off value 297.50 with sensitivity 85% and specificity 94.4%. Ferritin was increased in active disease   as compared to inactive disease and correlated with SLEDAI score and IgM-dsDNA. Thus, ferritin may be potential as an affordable and available marker of disease activity in pSLE during pandemic Covid-19.
铁蛋白是一种急性期反应物,在自身免疫性疾病中升高,包括系统性红斑狼疮(SLE)。然而,它们与疾病活动评分的相关性尚未得到证实。新冠肺炎大流行使儿童去医院接受SLE治疗的时间被推迟。本研究旨在评估血清铁蛋白与疾病活动性之间的相关性及其在新冠肺炎大流行期间儿童SLE(pSLE)患者发作筛查中的作用。这是在Saiful Anwar综合医院Malang进行的一项横断面研究。在2021年7月至2022年5月期间,对符合系统性狼疮国际合作诊所(SLICC)标准的儿科患者依次进行采样。使用SLE疾病活动指数2000(SLEDAI-2k)对所有患者进行访谈和疾病活动性评估。评分<4分被归类为无活动性疾病。通过标准实验室程序测量生化、血清学测试,包括疾病活动标志物和铁蛋白水平。用SPSS软件进行比较、相关和ROC曲线分析。共有38名女性pSLE参与了这项研究。患者的平均年龄为12.6岁 ± 3.02 年。活动性疾病的血清铁蛋白显著高于非活动性疾病,分别为84.50 ng/mL(68.00-151.75 ng/mL)和815.00 ng/mL(451.25-1570.00 ng/mL),p值<0.05具有统计学意义。血清铁蛋白与SLEDAI 2K(r = 0.890,p = 0.000)。血清铁蛋白与IgM抗双链DNA之间也存在相关性(r = 0.325,p = 0.046),但与其他实验室和血清学参数无关。在ROC曲线分析中,我们发现曲线下面积(AUC)为0.989,95%CI为0.964-1.014,p值为0000,截止值为297.50,敏感性为85%,特异性为94.4%。与非活动性疾病相比,活动性疾病中的铁蛋白增加,并与SLEDAI评分和IgM-dsDNA相关。因此,在新冠肺炎大流行期间,铁蛋白可能是pSLE中一种负担得起且可用的疾病活性标志物。
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Journal of Tropical Life Science
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