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Biophysical analysis of an oligomerization-attenuated variant of the Leishmania donovani dynamin-1-like protein 多诺瓦利什曼原虫动力蛋白-1样蛋白寡聚化减毒变体的生物物理分析。
IF 1.5 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-08-14 DOI: 10.1016/j.molbiopara.2025.111691
Ellen Wuyts , Ramasubramanian Sundaramoorthy , Lindsay Tulloch , Pieter Monsieurs , Thomas C. Eadsforth , Isabel Pintelon , Jean-Pierre Timmermans , Jean-Claude Dujardin , Malgorzata Anna Domagalska , Guy Caljon , Manu De Rycker , Vincent L.G. Postis , Susan Wyllie , Yann G.-J. Sterckx
Chemotherapy is a cornerstone in the battle against leishmaniasis, a neglected tropical disease caused by Leishmania parasites that affects millions worldwide. An alarming number of reports are describing treatment failure with currently available drugs, thereby explaining the dire need for the discovery of novel compounds, preferably with yet unexplored modes of action. In this respect L. donovani dynamin-1 like protein (LdoDLP1) is of interest as mutations in LdoDLP1 were recently shown to confer resistance to a new antileishmanial compound, suggesting it to be a potential drug target. Through a combination of biochemical, structural, and biophysical methods, we were able to show that wild-type LdoDLP1 has a strong inherent propensity to self-assemble into higher-order oligomers. Guided by structural modeling, a selection of nine point mutations (including resistance markers) were screened for oligomerization behavior to generate self-assembly impaired LdoDLP1 mutants that would occur in solution as dimers and/or tetramers. This led to the identification of a double mutant (G354D/R357S) that exhibits significantly altered and reduced, yet not completely abolished, oligomerization behavior. Further characterization of the LdoDLP1 G354D/R357S double mutant using small-angle X-ray scattering (SAXS) revealed that a fraction of the protein population occurs as a dimer in solution. Additionally, SAXS analysis experimentally confirmed that LdoDLP1, like other dynamin-like proteins, lacks the structurally defined pleckstrin homology (PH) domain of classical dynamins but instead possesses an intrinsically disordered B insert, grouping it among the dynamin-like proteins that play key roles in processes such as mitochondrial fission.
化疗是对抗利什曼病的基石。利什曼病是一种被忽视的热带疾病,由利什曼原虫引起,影响全世界数百万人。数量惊人的报告描述了目前可用药物的治疗失败,从而解释了迫切需要发现新的化合物,最好是具有尚未探索的作用模式。在这方面,L. donovani动力蛋白-1样蛋白(LdoDLP1)引起了人们的兴趣,因为LdoDLP1的突变最近被证明对一种新的抗利什曼原虫化合物具有耐药性,这表明它是一种潜在的药物靶点。通过生物化学、结构和生物物理方法的结合,我们能够证明野生型LdoDLP1具有很强的自组装成高阶低聚物的内在倾向。在结构建模的指导下,筛选了9个点突变(包括抗性标记)的寡聚化行为,以产生自组装受损的LdoDLP1突变体,这些突变体将以二聚体和/或四聚体的形式在溶液中发生。这导致了双突变体(G354D/R357S)的鉴定,该突变体表现出明显改变和减少的寡聚化行为,但并未完全消除。利用小角x射线散射(SAXS)对LdoDLP1 G354D/R357S双突变体的进一步表征显示,一部分蛋白质群体在溶液中以二聚体的形式出现。此外,SAXS分析实验证实,与其他动力蛋白样蛋白一样,LdoDLP1缺乏经典动力蛋白结构定义的pleckstrin同源(PH)结构域,而是具有内在无序的B插入,将其归类为动力蛋白样蛋白,在线粒体裂变等过程中发挥关键作用。
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引用次数: 0
Evaluation of the efficacy of Perillyl alcohol in the treatment of Eimeria tenella infection 紫苏醇治疗柔嫩艾美耳球虫感染的疗效评价
IF 1.5 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-07-30 DOI: 10.1016/j.molbiopara.2025.111690
Qingyang Song , Yihao Yu , Shujing Wang , Hongmei Li , Xiaomin Zhao , Ningning Zhao , Xiao Zhang
Recently, there has been an increased focus on the development of novel anti-parasitic drugs that exhibit both highly efficacy and low toxicity due to growing concerns associated with the widespread use of such drugs. Natural products have garnered significant interest owing to their diverse biological activities and minimal adverse effects. In this study, we assessed the anti-Eimeria tenella activity of four plant compounds belonging to the Lamiaceae family, namely Perillyl alcohol, Carvone, Menthone and Perilla aldehyde. Our in vitro experiments demonstrated that all four compounds, particularly Perillyl alcohol, exhibited potent inhibition against sporulation formation of E. tenella oocyst. Furthermore, our in vivo tests revealed that treatment with these four compounds at a dose of 200 mg/kg significantly reduced oocyst shedding as well as cecal lesions and weight loss caused by E. tenella infection, thereby demonstrating moderate anti-E. tenella activity. Notably, Perillyl alcohol displayed the highest efficacy against E. tenella with an anticoccidial index (ACI) value of 161.4. In summary, our findings indicate that these four compounds derived from the Lamiaceae family exhibit anti-E. tenella activity both in vitro and in vivo, with Perillyl alcohol displaying particularly robust inhibitory effects on E. tenella. It is worthy of further investigation to explore its mechanism of action and potential therapeutic applications.
最近,由于对新型抗寄生虫药物的广泛使用日益关注,人们越来越关注开发既高效又低毒性的新型抗寄生虫药物。天然产物因其多样的生物活性和最小的副作用而引起了人们的极大兴趣。本研究对紫苏醇、香芹酮、薄荷酮和紫苏醛四种紫苏科植物化合物的抗艾美耳球虫活性进行了研究。我们的体外实验表明,这四种化合物,特别是紫苏醇,对柔嫩芽胞杆菌卵囊的孢子形成具有有效的抑制作用。此外,我们的体内试验显示,以200 mg/kg的剂量用这四种化合物治疗可显著减少卵囊脱落、盲肠病变和由tenella感染引起的体重减轻,从而显示出适度的抗e。tenella活动。紫苏醇的抗球虫指数(ACI)为161.4,效果最好。综上所述,我们的研究结果表明,这四种来自Lamiaceae家族的化合物具有抗e。在体外和体内,紫苏醇对E. tenella表现出特别强大的抑制作用。其作用机制和潜在的治疗应用值得进一步研究。
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引用次数: 0
Molecular detection and population genetic diversity analysis of Theileria ovis in partial regions of Xinjiang, China 新疆部分地区鸡伊勒菌分子检测及群体遗传多样性分析
IF 1.4 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-07-16 DOI: 10.1016/j.molbiopara.2025.111689
Na Zhou , Meiling Wang , Xueqing Zhao , Abudukadier Mijiti , Wenying Dang , Licui Wen , Wenyu Shi , Lu Gan , Caishan Li , Bayinchahan Gailike
Ovine theileriosis is a disease caused by the genus Theileria (e.g., T. ovis, T. lestoquardi), preventing the sheep farming industry from developing, particularly in regions reliant on sheep for milk, meat, and associated economic benefits. However, there is limited information available on the epidemiological data and genetic diversity of T. ovis in Xinjiang. This study was conducted in May 2024 to investigate the molecular prevalence of T. ovis in sheep from five counties (Shaya, Wensu, Aketao, Keping, Awati) in Xinjiang. A total of 357 blood samples were screened for the presence of Theileria DNA through the amplification of the 18S rRNA gene using PCR, the genetic diversity among the chosen T. ovis sequences from geographical regions (including sequences in this study) was subsequently analyzed. BLAST analysis confirmed that the detected Theileria pathogen was T. ovis. Statistical results showed that the infection rate of T. ovis in sheep was 44.5 % (159/357). The highest infection rate was observed in Awati County, while the lowest was recorded in Shaya County. The prevalence exhibited significant variation among the sampling sites (χ² = 115.3, p < 0.05). To characterize the phylogenetic relationships within the detected Theileria populations, the sequenced T. ovis isolates were analyzed and found to be 96.6–99.8 % similar, showing a high degree of similarity to isolates from Turkey. Haplotype analysis further demonstrated that H1 constitutes the core haplotype (including sequences from Turkey, Iraq and Saudi Arabia), surrounded by derivative haplotype. To further investigate these haplotype distributions, population structure analysis revealed distinct genetic diversity patterns among groups, showing that genetic groups G1 and G4 had high haplotype diversity (Hd) but low nucleotide diversity (Pi), whereas G2 and G3 had low Hd and high Pi. In addition, Tajima's D<0 in all four T. ovis populations. These biological and genetic indices suggest that these populations are possibly undergoing expansion. Our results suggest that the protozoan parasitizing local sheep is T. ovis. Moreover, the local population of T. ovis is as rich in genetic diversity and population expansion as other populations in different geographical locations.
绵羊大肠杆菌病是一种由大肠杆菌属(例如,山羊绦虫、莱斯托quardi绦虫)引起的疾病,它阻碍了绵羊养殖业的发展,特别是在依赖绵羊获得奶、肉和相关经济利益的地区。但是,目前新疆地区的流行病学资料和遗传多样性资料有限。本研究于2024年5月在新疆沙雅、文苏、阿克陶、柯平、阿瓦提5个县(县)的绵羊中开展了羊链球菌分子流行病学调查。通过PCR扩增18S rRNA基因,筛选357份血液样本,检测其DNA的存在,并分析不同地理区域(包括本研究中的序列)所选择的T. ovis序列的遗传多样性。BLAST分析证实,检测到的病原菌为T. ovis。统计结果显示,绵羊中葡萄球菌感染率为44.5 %(159/357)。感染率最高的是阿瓦提县,最低的是沙雅县。各采样点的患病率差异有统计学意义(χ²= 115.3,p <; 0.05)。为了确定检测到的伊氏菌群之间的系统发育关系,我们对测序后的鸡伊氏菌分离株进行了分析,发现相似度为96.6-99.8 %,与土耳其分离株高度相似。单倍型分析进一步表明,H1构成核心单倍型(包括来自土耳其、伊拉克和沙特阿拉伯的序列),周围是衍生单倍型。为了进一步研究这些单倍型分布,群体结构分析揭示了不同群体间的遗传多样性模式,G1和G4遗传群具有高单倍型多样性(Hd)和低核苷酸多样性(Pi),而G2和G3遗传群具有低Hd和高Pi。此外,Tajima’s D在所有4个种群中均<0。这些生物学和遗传指标表明,这些种群可能正在扩大。我们的结果表明,当地绵羊的寄生原生动物是T. ovis。此外,在不同地理位置的其他种群中,当地的紫胸霉种群具有丰富的遗传多样性和种群扩张性。
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引用次数: 0
Characterization and immunogenicity analysis of glutathi- one S-transferase from Otodectes cynotis 耳细胞囊炎谷胱甘肽- 1 s -转移酶的特性及免疫原性分析。
IF 1.4 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-07-03 DOI: 10.1016/j.molbiopara.2025.111688
Hongbo Wang, Meilin Tan, Yinbo Gui, Xiaofang Wu, Maochuan Guo, Ran He
Otodectes cynotis (ear mite), the primary etiological agent of feline otitis externa, represents a significant veterinary concern due to its high prevalence and treatment challenges. Glutathione S-transferase (GST), a detoxifying and immunogenic enzyme in various parasites, is a potential molecular target for vaccine development. In this study, we cloned and heterologously expressed the GST gene from O. cynotis, confirmed its recombinant protein activity using 1-chloro-2,4-dinitrobenzene (CDNB) as a substrate, and determined its optimal enzymatic conditions (pH 8, 30 °C). Bioinformatic analysis revealed high sequence conservation with arthropod homologs, predicted functional domains, and identified several immunogenic B- and T-cell epitopes. Molecular docking with ethacrynic acid indicated stable binding, suggesting GST as a potential drug target. This study presents the first functional and immunogenic characterization of O. cynotis GST, suggesting its critical role in oxidative stress mitigation and drug detoxification, and supporting its potential as an anti-mite vaccine candidate.
耳螨是猫外耳炎的主要病原,由于其高患病率和治疗挑战,引起了兽医的重大关注。谷胱甘肽s -转移酶(GST)是多种寄生虫的解毒和免疫原性酶,是开发疫苗的潜在分子靶点。在本研究中,我们克隆并异源表达了O. cynotis的GST基因,以1-氯-2,4-二硝基苯(CDNB)为底物证实了其重组蛋白的活性,并确定了其最佳酶促条件(pH 8, 30°C)。生物信息学分析揭示了节肢动物同源物的高序列保守性,预测了功能域,并鉴定了几个免疫原性B和t细胞表位。与乙酸分子对接,结合稳定,提示GST是潜在的药物靶点。本研究首次提出了cynotis GST的功能和免疫原性特征,表明其在氧化应激缓解和药物解毒中起关键作用,并支持其作为抗螨候选疫苗的潜力。
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引用次数: 0
Identification and initial characterisation of Trypanosoma Cox17 copper metallochaperone 锥虫Cox17铜金属伴侣蛋白的鉴定与初步表征。
IF 1.4 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-06-18 DOI: 10.1016/j.molbiopara.2025.111687
Murtala Bindawa Isah, Mwenya Chitembusha Kwangu, J.P. Dean Goldring, Theresa H.T. Coetzer
Copper plays an essential role in organisms as a catalytic co-factor for key enzymes like cytochrome c oxidase. Copper importation, distribution and secretion is carried out by copper transport and copper-binding proteins known as copper chaperones. Cox17 is a chaperone that conveys copper to Cox11 and Sco1 for metalation of the CuB and CuA of Cox1 and Cox2 respectively in eukaryotes. Cox17 from Trypanosoma brucei brucei and T. congolense were recombinantly expressed and affinity purified as MBP-fusion proteins. An ascorbic acid oxidation assay, a BCA-release assay and an in vivo growth inhibition assay confirmed the presence of copper bound to the proteins. Trypanosomal Cox17 and other copper-binding proteins are expressed at higher levels in the insect procyclic stage where cytochrome c oxidase is active, compared to the bovine bloodstream forms. In silico docking models suggests possible interaction partners for Cox17.
铜作为细胞色素c氧化酶等关键酶的催化辅助因子在生物体中起着至关重要的作用。铜的输入、分布和分泌是通过铜转运和铜结合蛋白(即铜伴侣蛋白)进行的。Cox17是一种伴侣蛋白,它将铜传递给Cox11和Sco1,分别用于真核生物Cox1和Cox2的CuB和CuA的金属化。重组表达了来自布氏锥虫和刚果锥虫的Cox17,并将其纯化为mbp融合蛋白。抗坏血酸氧化试验、bca释放试验和体内生长抑制试验证实了铜与蛋白质结合的存在。与牛的血流形式相比,在细胞色素c氧化酶活跃的昆虫前环阶段,锥虫体Cox17和其他铜结合蛋白的表达水平更高。计算机对接模型显示了Cox17可能的相互作用伙伴。
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引用次数: 0
Calcium/calmodulin-dependent protein kinase II in Schistosoma: Relation to praziquantel action and resistance 血吸虫钙/钙调素依赖性蛋白激酶II:与吡喹酮作用和抗性的关系
IF 1.4 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-05-23 DOI: 10.1016/j.molbiopara.2025.111686
Iman F. Abou-El-Naga
Calcium/calmodulin-dependent protein kinase II (CaMKII) performs diverse essential functions through integrating a range of calcium signals. In Schistosoma, two Calmodulin (CaM) genes are characterized. CaMKII exhibits distinct expression patterns across the developmental stages of the parasite. Its significance lies in sustaining Schistosoma survival and maintaining calcium homeostasis. As it is a calcium sensing protein, its function is closely related to the efficacy of praziquantel, the mainstay drug against schistosomiasis. The relationship between CaMKII and praziquantel involves several potential factors. Praziquantel induces an increased calcium influx into Schistosoma that binds CaM and activates CaMKII, which in turn mitigates the effect of the drug and potentially contributes to praziquantel resistance in several ways. By maintaining calcium homeostasis, CaMKII opposes the surge in calcium influx induced by praziquantel. It modulates voltage-gated calcium channels and reduces calcium influx. It also inhibits ryanodine receptors and inositol triphosphate receptors, thus preventing the release of calcium from the sarcoplasmic/endoplasmic reticulum. CaMKII activates nuclear factor-κB and subsequently activates sarco/endoplasmic reticulum calcium-ATPase (SERCA), which increases calcium uptake into the sarcoplasmic/endoplasmic reticulum and decreases cytosolic calcium. Nuclear factor-κB, activated by CaMKII may lead to up-regulation of P-glycoprotein, which facilitates praziquantel efflux. CaMKII also activates calcineurin that inhibits SERCA. Given its pivotal role in Schistosoma homeostasis and survival, CaMKII emerges as a promising target for novel anthelmintic therapies, and its modulation might enhance the efficacy of praziquantel.
钙/钙调素依赖性蛋白激酶II (CaMKII)通过整合一系列钙信号发挥多种基本功能。在血吸虫中,两个钙调素(CaM)基因被鉴定。CaMKII在寄生虫的不同发育阶段表现出不同的表达模式。其意义在于维持血吸虫存活和维持钙稳态。由于它是一种钙敏感蛋白,其功能与吡喹酮的疗效密切相关,吡喹酮是血吸虫病的主要药物。CaMKII与吡喹酮之间的关系涉及几个潜在因素。吡喹酮诱导更多的钙流入血吸虫,结合CaM并激活CaMKII,这反过来减轻了药物的作用,并可能以多种方式促进吡喹酮耐药性。通过维持钙稳态,CaMKII对抗吡喹酮引起的钙流入激增。它调节电压门控钙通道,减少钙流入。它还能抑制红嘌呤受体和肌醇三磷酸受体,从而阻止钙从肌浆/内质网释放。CaMKII激活核因子-κB,随后激活肌浆/内质网钙atp酶(SERCA),增加肌浆/内质网钙摄取,减少胞质钙。CaMKII激活核因子-κB可导致p-糖蛋白上调,促进吡喹酮外排。CaMKII也激活钙调磷酸酶,抑制SERCA。鉴于其在血吸虫体内平衡和生存中的关键作用,CaMKII成为新型驱虫药治疗的一个有希望的靶点,其调节可能增强吡喹酮的疗效。
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引用次数: 0
Biological and physiological changes of Biomphalaria glabrata infected by Heterorhabditis bacteriophora 光秃生物phalaria感染噬菌体异habditis后的生物学和生理变化
IF 1.4 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-05-14 DOI: 10.1016/j.molbiopara.2025.111679
Vinícius Menezes Tunholi-Alves , Victor Menezes Tunholi , Ludimila Santos Amaral , Natânia do Carmo Sperandio , Lorena Souza Castro Altoé , Melissa Carvalho Machado do Couto-Chambarelli , Caio Márcio de Oliveira Monteiro , Isabella Vilhena Freire Martins
The present study investigated for the first time the influence of exposure time (24 or 48 h) of Biomphalaria glabrata (Gastropoda: Pulmonata) to Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae), strain HP88, on some physiological and reproductive parameters of the host mollusk. Throughout the experiment, intense glycogenolysis was observed in both exposed groups, which was more accentuated in the digestive gland. This change was accompanied by a significant increase in the free glucose content in the exposed snails, indicating that H. bacteriophora infection induces the breakdown of host glycemic homeostasis. In parallel, significant variations in lactate dehydrogenase activity in the hemolymph of B. glabrata exposed to entomopathogenic nematodes were observed, confirming the transition from aerobic to anaerobic metabolism in the hosts. This physiological scenario contributed to the establishment of the parasitic castration process in this interface, compromising the reproductive performance of host snails, suggesting the use of H. bacteriophora HP88 as a potential alternative for control of B. glabrata.
本文首次研究了腹足目:肺足目(Gastropoda: Pulmonata)暴露于菌株Heterorhabditis bacteriophora (rhabdida: heterorhabditida: heterorhabdititidae) HP88的时间(24或48 h)对寄主软体动物某些生理和生殖参数的影响。在整个实验过程中,两个暴露组均观察到剧烈的糖原溶解,且在消化腺中更为明显。这种变化伴随着暴露的蜗牛体内游离葡萄糖含量的显著增加,表明嗜杆菌感染诱导了宿主血糖稳态的破坏。与此同时,暴露于昆虫病原线虫的光斑拟南蝽血淋巴中乳酸脱氢酶活性也发生了显著变化,证实了宿主的代谢从有氧向无氧的转变。这一生理情景促成了该界面寄生去势过程的建立,影响了寄主蜗牛的繁殖性能,表明利用H. bacteriophora HP88作为控制光斑螺的潜在替代方案。
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引用次数: 0
The role of complement system in a gerbil model of cutaneous leishmaniasis 补体系统在沙鼠皮肤利什曼病模型中的作用。
IF 1.4 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-04-01 DOI: 10.1016/j.molbiopara.2025.111678
Baycan Mor , Arzu Gormez , Berna Demırcı
Leishmania species are intracellular protozoans responsible for causing both cutaneous and visceral infections. In recent years, the prevalence of leishmaniasis, a systemic and chronic disease, has been on the rise. Complement pathway mechanisms, part of the immune response of host organisms against Leishmania species, have not been fully revealed in leishmaniasis, which is very important for public health. This study aimed to explore the role of the complement system, an integral part of the immune response to Leishmania infections, in gerbil (Meriones unguiculatus) models of cutaneous leishmaniasis. This was achieved by assessing the expression levels of complement system genes (MBL-1, MBL-2, C2, and C3) and quantifying the protein levels of MBL-1, C2, and C3. Additionally, the study aimed to conduct biochemical tests, specifically measuring GSH and MDA levels, to detect oxidative stress in response to infection in gerbils. Finally, hematological analyses were performed to evaluate leukocyte counts in the blood. The expression of complement system genes and some complement system proteins were significantly increased in infected gerbils. Oxidative stress was evident, as indicated by reduced GSH levels and increased MDA levels. Additionally, a significant rise in leukocyte counts was observed as a consequence of the infection. The study concluded that complement system pathways are activated in cutaneous leishmaniasis infections. It was also determined that a thorough evaluation of genomic, proteomic, and immunopathological mechanisms is essential for understanding the pathogenesis of the disease.
利什曼原虫属细胞内原生动物,可引起皮肤和内脏感染。近年来,利什曼病(一种全身性慢性疾病)的流行率一直在上升。补体途径机制是宿主机体对利什曼原虫免疫反应的一部分,在利什曼病中尚未完全揭示,这对公共卫生具有重要意义。本研究旨在探讨补体系统在沙鼠皮肤利什曼病模型中的作用,补体系统是对利什曼感染免疫反应的一个组成部分。这是通过评估补体系统基因(MBL-1、MBL-2、C2和C3)的表达水平和量化MBL-1、C2和C3的蛋白水平来实现的。此外,该研究旨在进行生化测试,特别是测量谷胱甘肽和丙二醛水平,以检测沙鼠对感染的氧化应激反应。最后,进行血液学分析以评估血液中的白细胞计数。感染沙鼠体内补体系统基因及部分补体系统蛋白的表达显著升高。氧化应激明显,GSH水平降低,MDA水平升高。此外,白细胞计数显著上升被观察到作为感染的结果。该研究得出结论,补体系统途径在皮肤利什曼病感染中被激活。研究人员还认为,全面评估基因组学、蛋白质组学和免疫病理机制对于了解该病的发病机制至关重要。
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引用次数: 0
Metronidazole induces prostaglandin E2 formation via arachidonic acid production in protozoan parasite Giardia lamblia 甲硝唑诱导原生动物贾第鞭毛虫产生花生四烯酸形成前列腺素E2。
IF 1.4 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-03-16 DOI: 10.1016/j.molbiopara.2025.111676
Rituparna Sarkar , Sanjib Kumar Sardar , Ajanta Ghosal , Tapas Haldar , Koushik Das , Arjun Ghosh , Akash Prasad , Yumiko Saito-Nakano , Shanta Dutta , Tomoyoshi Nozaki , Sandipan Ganguly
The causative agent of giardiasis in human and animals is the amitochondriate Giardia lamblia. We observed that exposing Giardia trophozoites to MTZ led to an increase in lipid peroxidation compared to the control group, which was expressed in terms of menadione production as it is the marker for lipo-peroxidation. Oxidative stress generated by reactive nitrogen species and peroxidation of membrane phospholipids are positively correlated with the enhanced PLA2 activity in several organisms to produce arachidonic acid (AA). Our data suggested Giardia produces a unique 56 kDa dimeric enzyme called Phospholipase B (gPLB) in contrast to higher eukaryotes which was responsible for the production of intracellular free AA. This free AA either reacylates to the cell membrane or deacylates to further produce prostaglandins. In normal un-induced controlled trophozoites the membrane reacylation process was dominant due the higher level of acyle CoA synthase (ACS) expression over the time. However, under the oxidative stressed condition the intracellular ACS expression was down regulated. This led to the increase in deacylation process. When AA deacylation becomes dominant over AA reacylation in cells, the free AA accumulates intracellularly. One of the lipid autacoids, derived from AA is prostaglandin2 (PGE2). Oxidative stress generated by reactive nitrogen species in trophozoites increased the PGE2 production via prostaglandin synthase over the time with respect to the controlled one.
人类和动物贾第虫病的病原体是带线粒体的贾第虫。我们观察到,与对照组相比,将贾第鞭毛虫滋养体暴露于MTZ导致脂质过氧化增加,这表现为甲萘醌的产生,因为它是脂质过氧化的标志。活性氮产生的氧化应激和膜磷脂的过氧化与几种生物产生花生四烯酸(AA)的PLA2活性增强呈正相关。我们的数据表明贾第鞭毛虫产生一种独特的56kDa二聚体酶,称为磷脂酶B (gPLB),与高级真核生物相比,它负责产生细胞内游离AA。这种游离的AA要么再乙酰化到细胞膜上,要么去乙酰化进一步产生前列腺素。在正常的未诱导的控制滋养体中,由于随着时间的推移,环辅酶a合成酶(ACS)的表达水平较高,膜再酰基化过程占主导地位。而在氧化应激条件下,细胞内ACS表达下调。这导致了去酰化过程的增加。当细胞内AA去酰化作用大于AA再酰化作用时,游离AA在细胞内积累。其中一种脂质类自身素,来源于AA是前列腺素2 (PGE2)。活性氮在滋养体中产生的氧化应激随着时间的推移增加了前列腺素合成酶产生的PGE2。
{"title":"Metronidazole induces prostaglandin E2 formation via arachidonic acid production in protozoan parasite Giardia lamblia","authors":"Rituparna Sarkar ,&nbsp;Sanjib Kumar Sardar ,&nbsp;Ajanta Ghosal ,&nbsp;Tapas Haldar ,&nbsp;Koushik Das ,&nbsp;Arjun Ghosh ,&nbsp;Akash Prasad ,&nbsp;Yumiko Saito-Nakano ,&nbsp;Shanta Dutta ,&nbsp;Tomoyoshi Nozaki ,&nbsp;Sandipan Ganguly","doi":"10.1016/j.molbiopara.2025.111676","DOIUrl":"10.1016/j.molbiopara.2025.111676","url":null,"abstract":"<div><div>The causative agent of giardiasis in human and animals is the amitochondriate <em>Giardia lamblia.</em> We observed that exposing <em>Giardia</em> trophozoites to MTZ led to an increase in lipid peroxidation compared to the control group, which was expressed in terms of menadione production as it is the marker for lipo-peroxidation. Oxidative stress generated by reactive nitrogen species and peroxidation of membrane phospholipids are positively correlated with the enhanced PLA2 activity in several organisms to produce arachidonic acid (AA). Our data suggested <em>Giardia</em> produces a unique 56 kDa dimeric enzyme called Phospholipase B (gPLB) in contrast to higher eukaryotes which was responsible for the production of intracellular free AA. This free AA either reacylates to the cell membrane or deacylates to further produce prostaglandins. In normal un-induced controlled trophozoites the membrane reacylation process was dominant due the higher level of acyle CoA synthase (ACS) expression over the time. However, under the oxidative stressed condition the intracellular ACS expression was down regulated. This led to the increase in deacylation process. When AA deacylation becomes dominant over AA reacylation in cells, the free AA accumulates intracellularly. One of the lipid autacoids, derived from AA is prostaglandin2 (PGE2). Oxidative stress generated by reactive nitrogen species in trophozoites increased the PGE2 production via prostaglandin synthase over the time with respect to the controlled one.</div></div>","PeriodicalId":18721,"journal":{"name":"Molecular and biochemical parasitology","volume":"262 ","pages":"Article 111676"},"PeriodicalIF":1.4,"publicationDate":"2025-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143657733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Influences of parasitic stress on the health condition of African Catfish (Clarias gariepinus): Biochemical and histopathological alterations 寄生胁迫对非洲鲶鱼健康状况的影响:生化和组织病理学改变。
IF 1.4 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-03-09 DOI: 10.1016/j.molbiopara.2025.111677
Hadeer Abd El-hak Rashed , Nahla S. El-Shenawy , Nadia A. El-Fahla
This study investigates the prevalence, severity, and impacts of parasitic infestations in Clarias gariepinus. Additionally, the study assesses the detrimental impacts of parasite infestation on the health condition of affected catfish, focusing on biochemical and histopathological alterations. A total of 160 fish were sampled from local markets. Parasitological examinations involved the dissection of key organs from each fish. The organs were processed and examined microscopically for parasites identified based on morphometric characteristics. Parasitological indices such as prevalence, mean intensity, and abundance were calculated. Fish blood and liver samples were collected to assess hematological and biochemical parameters. Microscopic and ultrastructural examinations identified the gills and liver as highly infected organs, so they were utilized for transmission electron microscopy (TEM). Analysis of catfish tissues unveiled the existence of Cyathocotylid sp. and Prohemistomum vivax, across all organs with dominance noted in the liver, emphasizing their pathogenic significance and notable ability to invade and establish within multiple organs or the immunocompromised response of the host. Meanwhile, Centrocestus formosanus and Quadriacanthus aegyptiacus were exclusively detected in the gills, with an overall parasitic infection rate of 60 %. The present study is one of the few studies documenting Centrocestus sp. in catfish which reflects its ability to spread in new hosts and environments. A novel morphological dimension was recorded for the recovered metacercariae. The hematological, along with the identified lesions from light histological and TEM examinations in heavily infected catfish, indicate the detrimental impact of parasite infiltration on fish health status. Besides the biochemical biomarkers were significantly (p ≤ 0.05) affected by increasing the degree of infection. This study underscores the profound influence of parasitic infestations on the health of C. gariepinus, emphasizing the urgent need for effective management strategies in aquaculture to mitigate these effects, the spread of new pathogens, and ensure the sustainability and productivity of catfish farming. By integrating parasitological, morphological, histopathological, and biochemical analyses, this research provides valuable insights that contribute to better health management strategies in aquaculture and a deeper understanding of parasite biology.
本研究调查了加里宾Clarias gariepinus寄生虫侵染的流行程度、严重程度和影响。此外,该研究还评估了寄生虫侵染对受感染鲶鱼健康状况的有害影响,重点是生化和组织病理学改变。本署在本港市场共抽取160条鱼样本。寄生虫学检查包括解剖每条鱼的关键器官。对器官进行处理,并在显微镜下检查根据形态特征确定的寄生虫。计算流行率、平均强度、丰度等寄生虫学指标。采集鱼类血液和肝脏样本,评估血液学和生化参数。显微镜和超微结构检查发现鳃和肝脏是高度感染的器官,因此用透射电子显微镜(TEM)观察。对鲶鱼组织的分析揭示了Cyathocotylid sp.和Prohemistomum vivax的存在,在所有器官中都有,在肝脏中有优势,强调了它们的致病意义和在多个器官中入侵和建立的显着能力或宿主的免疫低下反应。同时,在鳃中仅检出台湾Centrocestus formosanus和埃及Quadriacanthus aegyptiacus,总体寄生虫感染率为60%。本研究是为数不多的记录鲶鱼中Centrocestus sp.的研究之一,反映了它在新宿主和新环境中的传播能力。在恢复的囊蚴中记录了一种新的形态维度。血液学,以及在严重感染的鲶鱼的光组织学和透射电镜检查中发现的病变,表明寄生虫渗透对鱼类健康状况的有害影响。此外,随着感染程度的增加,各生化指标均受到显著影响(p≤0.05)。本研究强调了寄生虫侵染对鲶鱼健康的深远影响,强调了迫切需要有效的水产养殖管理策略来减轻这些影响,防止新病原体的传播,确保鲶鱼养殖的可持续性和生产力。通过整合寄生虫学、形态学、组织病理学和生化分析,本研究为水产养殖中更好的健康管理策略和更深入地了解寄生虫生物学提供了有价值的见解。
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Molecular and biochemical parasitology
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