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Loop-mediated isothermal amplification (LAMP): A promising tool for rapid, point-of-care diagnosis of parasitic diseases in low-income countries 环介导等温扩增(LAMP):低收入国家快速、即时诊断寄生虫病的一种有前途的工具。
IF 1.5 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-10-10 DOI: 10.1016/j.molbiopara.2025.111704
Kevin Polpitiya, Madhavi Hewadikaram
Parasitic diseases pose a major burden throughout the globe, especially in low-income countries, which are disproportionately affected due to factors such as overpopulation, vector-control challenges, financial difficulties, etc. Because access to advanced diagnostic technologies is limited in low-income regions, early and accurate diagnosis, which is critical in preventing diseases from progressing to their fatal stages, is not possible. Methods such as polymerase chain reaction (PCR) are often impractical in such regions due to their complexity and high running costs. Loop-mediated isothermal amplification (LAMP) is a promising alternative diagnostic technology that amplifies and detects a specific DNA sequence from a sample. Its features, such as high sensitivity and specificity, affordability, simplicity and versatility, make it an attractive alternative for poorer countries. Moreover, LAMP operates under isothermal conditions, thereby reducing the need for expensive equipment. This review evaluates the applicability of LAMP to be used as an alternative POC diagnostic method for parasitic infections by comparing it with routinely used diagnostic methods. Furthermore, several emerging trends in LAMP technology, such as multiplex LAMP, reverse transcription LAMP, microfluidic chip-incorporated LAMP, digital LAMP, and single nucleotide polymorphism LAMP, are discussed, further illustrating the potential of improving the diagnostic accuracy and affordability of LAMP. The need for cost-effective and accurate diagnostic tools is critical, particularly in regions where parasitic diseases are most prevalent. LAMP represents a viable solution to reduce the burden of parasitic diseases through improved diagnostic capabilities.
寄生虫病在全球范围内构成重大负担,特别是在低收入国家,由于人口过剩、病媒控制挑战、财政困难等因素,这些国家受到的影响尤为严重。由于在低收入地区获得先进诊断技术的机会有限,因此不可能进行早期和准确的诊断,而这对防止疾病发展到致命阶段至关重要。聚合酶链反应(PCR)等方法由于其复杂性和高运行成本,在这些地区往往不切实际。环介导等温扩增(LAMP)是一种很有前途的替代诊断技术,可以从样品中扩增和检测特定的DNA序列。它的特点,如高灵敏度和特异性、可负担性、简单性和多功能性,使其成为较贫穷国家的一个有吸引力的选择。此外,LAMP在等温条件下工作,从而减少了对昂贵设备的需求。本文通过与常规诊断方法的比较,评价LAMP作为一种可替代的POC寄生虫感染诊断方法的适用性。此外,还讨论了LAMP技术的几个新兴趋势,如多路LAMP、逆转录LAMP、微流控芯片LAMP、数字LAMP和单核苷酸多态性LAMP,进一步说明了提高LAMP诊断准确性和可负担性的潜力。需要具有成本效益和准确的诊断工具至关重要,特别是在寄生虫病最流行的区域。LAMP是通过改进诊断能力来减轻寄生虫病负担的可行解决办法。
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引用次数: 0
Surface molecules of Leishmania: From virulence determinants to therapeutic and vaccine targets 利什曼原虫表面分子:从毒力决定因素到治疗和疫苗靶点。
IF 1.5 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-09-30 DOI: 10.1016/j.molbiopara.2025.111702
Túlio Custódio Reis , Ana Clara Lunardi Yagi , Ana Laura Dias Ramos , Angela Maria Arenas Velásquez , Natália Caroline Costa Coelho , Márcia A.S. Graminha
Leishmaniasis is a group of neglected tropical diseases (NTDs) caused by protozoa of the genus Leishmania that affect vulnerable populations in tropical and subtropical regions. The disease manifests in cutaneous, mucocutaneous, and visceral clinical forms. This major public health disease presents high morbidity, and despite the global impact of leishmaniasis, there are few therapeutic options available and no currently licensed human vaccines. Besides, the available therapeutic agents are associated with high toxicity and treatment failure. These limitations highlight the importance of identifying new therapeutic targets, which will contribute to the development of more effective, safer and shorter treatment options. In this context, surface molecules of Leishmania emerge as attractive therapeutic targets due to their roles in host cell adhesion, immune evasion, and intracellular survival. In addition to their translational potential for drug discovery and vaccine development, these surface molecules are key virulence factors that play central roles in parasite biology and disease pathogenesis. Understanding their structure and function is essential not only for elucidating mechanisms of host–parasite interaction, but also for identifying novel therapeutic and prophylactic strategies. Importantly, molecules such as GP63 (a major surface metalloprotease), LPG (lipophosphoglycan), and KMP-11 (kinetoplastid membrane protein 11) combine essential biological functions with demonstrated immunogenic properties, making them promise as targets for both chemotherapeutic and prophylactic interventions. This review aims to explore the structural and functional characteristics of major surface virulence factors in Leishmania, highlighting their roles in the parasite–host interaction and discussing their translational potential for therapeutic and vaccine development.
利什曼病是一组被忽视的热带病(NTDs),由利什曼属原虫引起,影响热带和亚热带地区的脆弱人群。本病临床表现为皮肤、粘膜和内脏。这一重大公共卫生疾病发病率很高,尽管利什曼病影响全球,但可用的治疗选择很少,目前也没有获得许可的人用疫苗。此外,现有的治疗药物具有高毒性和治疗失败的特点。这些限制突出了确定新的治疗靶点的重要性,这将有助于开发更有效、更安全、更短的治疗方案。在这种情况下,利什曼原虫的表面分子由于其在宿主细胞粘附、免疫逃避和细胞内存活中的作用而成为有吸引力的治疗靶点。除了它们在药物发现和疫苗开发中的翻译潜力外,这些表面分子是在寄生虫生物学和疾病发病机制中发挥核心作用的关键毒力因子。了解它们的结构和功能不仅对阐明宿主-寄生虫相互作用的机制,而且对确定新的治疗和预防策略至关重要。重要的是,GP63(一种主要的表面金属蛋白酶)、LPG(脂磷酸聚糖)和KMP-11(动质体膜蛋白11)等分子结合了基本的生物学功能和已证实的免疫原性,使它们有望成为化疗和预防性干预的靶点。本文旨在探讨利什曼原虫主要表面毒力因子的结构和功能特征,强调它们在寄生虫-宿主相互作用中的作用,并讨论它们在治疗和疫苗开发中的转化潜力。
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引用次数: 0
Computational investigation of mutations in PfCRT and PfDHFR proteins for emerging resistance of Plasmodium falciparum to antimalarial drugs 恶性疟原虫对抗疟药物产生耐药性的PfCRT和PfDHFR蛋白突变的计算研究
IF 1.5 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-09-05 DOI: 10.1016/j.molbiopara.2025.111700
Sushruta Ghosh , Deepesh Joshi , Chandra Sekar Ponnusamy , Bhavani Sridharan , Mahesh Velusamy
The emergence of multidrug resistance in Plasmodium falciparum poses a serious threat to antimalarial treatment, particularly with growing resistance to artemisinin-based combination therapies (ACTs) and partner drugs like piperaquine. Mutations in key proteins, such as PfCRT (P. falciparum chloroquine resistance transporter) and PfDHFR (P. falciparum dihydrofolate reductase), play a critical role in this resistance. Understanding these molecular mechanisms is essential for the development of effective antimalarial therapies. This study aimed to investigate the structural and functional impact of polymorphisms on drug-target interactions and resistance mechanisms in P. falciparum. Molecular docking and molecular dynamics (MD) simulations were performed to analyze interactions of the mutated PfCRT and PfDHFR proteins with nine antimalarial drugs, including piperaquine. The PfCRT-K76A piperaquine complex strong binding affinity (-9.5 kcal/mol) with moderate structural deviation (0.970 ± 0.202 nm) and greater solvent accessibility (246.01 ± 6.135 nm²), suggesting favourable binding conditions. The PfDHFR-N51I–piperaquine complex showed even stronger binding (-10.8 kcal/mol) but higher structural fluctuation (RMSD: 4.491 ± 1.462 nm) and increased compactness (1.861 ± 0.029 nm), which may reflect restricted ligand accommodation and possible resistance. Overall, the findings provide valuable insights into how PfCRT and PfDHFR mutations contribute to drug resistance and establish a foundation for designing more effective antimalarial strategies. Future research should integrate experimental validation and explore additional resistance-associated mutations to develop targeted therapies for combating multidrug-resistant P. falciparum.
恶性疟原虫多药耐药性的出现对抗疟治疗构成严重威胁,特别是对以青蒿素为基础的联合疗法和哌喹等伴用药的耐药性日益增强。关键蛋白的突变,如PfCRT(恶性疟原虫氯喹耐药转运蛋白)和PfDHFR(恶性疟原虫二氢叶酸还原酶),在这种耐药性中起关键作用。了解这些分子机制对于开发有效的抗疟疾疗法至关重要。本研究旨在探讨恶性疟原虫基因多态性对药物-靶点相互作用和耐药机制的结构和功能影响。通过分子对接和分子动力学(MD)模拟分析突变的PfCRT和PfDHFR蛋白与包括哌喹在内的9种抗疟疾药物的相互作用。PfCRT-K76A哌喹配合物结合亲和力强(-9.5 kcal/mol),结构偏差适中(0.970 ± 0.202 nm),溶剂可溶性较好(246.01 ± 6.135 nm²),结合条件良好。pfdhfr - n51i -哌喹配合物的结合强度更强(-10.8 kcal/mol),但结构波动更大(RMSD: 4.491 ± 1.462 nm),致密度更高(1.861 ± 0.029 nm),这可能反映了配体调节受限和可能的耐药性。总的来说,这些发现为PfCRT和PfDHFR突变如何促进耐药性提供了有价值的见解,并为设计更有效的抗疟疾策略奠定了基础。未来的研究应整合实验验证并探索更多的耐药相关突变,以开发针对耐多药恶性疟原虫的靶向治疗方法。
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引用次数: 0
Decoding Leishmania in equines: A comparative analysis of molecular targets 解码马的利什曼原虫:分子靶点的比较分析。
IF 1.5 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-09-03 DOI: 10.1016/j.molbiopara.2025.111699
Muhammad Adnan Sabir Mughal , Muhammad Kasib Khan , He Lan , Rao Zahid Abbas , Muhammad Imran , Zaheer Abbas , Muhammad Shahid Mehmood , Sultan Ali
Parasitic diseases caused by Leishmania spp. create considerable health concerns in animals, resulting in a considerable financial impact. They causes a complex infection in equines, affecting weight gain, skin, liver, and spleen. To date, there is a lack of reports on the occurrence of Leishmania in equines in Southern Punjab, Pakistan, highlighting the need for molecular epidemiological surveillance. The current study focused on determining the prevalence of Leishmania in the equine population from District Rahim Yar Khan, Southern Punjab, Pakistan, through amplification of mitochondrial (Cytochrome b) and nuclear (18S rRNA) genes of the parasite. For this purpose, a total of 384 equine - i.e. horses, mules, and donkeys - blood specimens, determined by calculation of the sample size formula, were obtained from District Rahim Yar Khan. The parasite was examined through the Microhematocrit method under the microscope. Leishmania was detected from the buffy coat layer after centrifugation of blood-filled microhematocrit tubes. To detect and characterize Leishmania spp.at the molecular level, DNA extraction from blood samples was carried out using standardized commercial kits, followed by PCR amplification. Information on potential risk factors was gathered through a structured questionnaire. The overall prevalence of Leishmania infection was observed to be 2.1 % via microscopy and 7.3 % and 8.8 % by amplification of the 18S rRNA and Cytochrome b genes using molecular methods. A significantly higher infection percentage was observed in female animals compared to males, and in older and underweight animals compared to younger and healthier ones. Additionally, the infection was non-significantly (P ≥ 0.05) more prevalent in gestating, non-dewormed, symptomatic, and poor body condition animals. Phylogenetic and sequence analyses confirmed that the identified gene sequences clustered within the Leishmania (Leishmania) infantum clade, consistent with strains reported in different animal hosts from various regions. In conclusion, the nuclear gene, i.e., 18S rRNA proved to be a more sensitive molecular marker for detecting Leishmania infection in equines compared to the mitochondrial gene, i.e., Cytochrome b.
利什曼原虫引起的寄生虫病在动物中造成相当大的健康问题,造成相当大的经济影响。它们会引起马的复杂感染,影响体重增加、皮肤、肝脏和脾脏。迄今为止,缺乏关于巴基斯坦南旁遮普省马中利什曼原虫发生的报告,这突出表明需要进行分子流行病学监测。目前的研究重点是通过扩增利什曼原虫的线粒体(细胞色素b)和细胞核(18S rRNA)基因,确定利什曼原虫在巴基斯坦南部旁遮普省拉希姆亚尔汗区马种群中的流行情况。为此目的,通过计算样本量公式,从拉希姆亚尔汗区获得了384个马(即马、骡子和驴)的血液标本。显微镜下用微血细胞比容法检查寄生虫。在充满血液的微红细胞压积管离心后,从肉皮层检测利什曼原虫。为了在分子水平上检测和表征利什曼原虫,使用标准化的商用试剂盒从血液样本中提取DNA,然后进行PCR扩增。通过结构化问卷收集潜在风险因素的信息。显微镜下观察利什曼原虫感染的总患病率为2.1%,分子法扩增18S rRNA和细胞色素b基因为7.3%和8.8%。与雄性动物相比,雌性动物的感染率要高得多,与年轻健康的动物相比,老年和体重不足的动物的感染率要高得多。妊娠期、未脱虫、有症状、体况较差的动物感染发生率无统计学意义(P≥0.05)。系统发育和序列分析证实,所鉴定的基因序列聚集在利什曼原虫(利什曼原虫)幼支中,与来自不同地区的不同动物宿主中报道的菌株一致。综上所述,与线粒体基因细胞色素b相比,核基因18S rRNA是检测马利什曼原虫感染的更敏感的分子标记。
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引用次数: 0
Pepscan and bioinformatic strategies for identification of potential B-cell epitopes for a peptide-based vaccine for tick control 蜱虫控制肽基疫苗潜在b细胞表位鉴定的Pepscan和生物信息学策略
IF 1.5 4区 医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2025-08-28 DOI: 10.1016/j.molbiopara.2025.111692
Vinícius Andrade-Silva , Jéssica Waldman , Maria Aparecida Juliano , Lucas Tirloni , Itabajara da Silva Vaz Jr.
The cattle tick Rhipicephalus microplus poses a major problem to the livestock industry worldwide, with acaricides resistance presenting an increasing challenge. On other hand, vaccination has been suggested as a better strategy for tick control, and peptide-based vaccines could be developed to target multiple tick antigens. Nevertheless, there are still limitations to the identification of epitopes in tick candidate antigens, as the bioinformatics tools currently available were developed almost exclusively based on mammalian genomes. Therefore, improving the performance of B-cell epitope predictor algorithms is essential to achieve an effective multi-epitope vaccine for tick control. The aim of this study was to reduce costs and increase the efficacy in identifying epitopes in tick antigens. We first evaluated the performance of B-cell epitope predictor algorithms in replicating the results of an in vitro epitope mapping result for the tick salivary serpin RmS-17 as a “benchmark”. Then the algorithm with the best performance was employed to predict epitopes for the tick salivary serpin RmS-6, and we screened the candidate epitopes based on predictions that were close to the reactive center loop (RCL), the region of the serpin that interacts with the target protease. Antibodies raised against p1RmS-6 and p3RmS-6 neutralize RmS-6 activity. Using this strategy, we were able to adjust an in silico algorithm predictor based on a Pepscan result to identify epitopes in another serpin. Our strategy offers a cost-effective way to identify neutralizing epitopes in serpins. Furthermore, this strategy can be applied to identify epitopes in serpins and other proteins from other tick species, potentially leading to the development of a peptide-based anti-tick vaccine.
牛蜱对世界范围内的畜牧业构成了一个重大问题,对杀螨剂的抗性提出了越来越大的挑战。另一方面,疫苗接种被认为是控制蜱虫的更好策略,肽类疫苗可以针对多种蜱虫抗原开发。然而,蜱候选抗原表位的鉴定仍然存在局限性,因为目前可用的生物信息学工具几乎完全是基于哺乳动物基因组开发的。因此,提高b细胞表位预测算法的性能对于实现有效的蜱虫多表位疫苗至关重要。本研究的目的是降低成本,提高识别蜱抗原表位的效率。我们首先评估了b细胞表位预测算法在复制蜱唾液丝氨酸蛋白RmS-17体外表位定位结果作为“基准”方面的性能。然后采用性能最佳的算法预测蜱唾液蛇形蛋白RmS-6的表位,并根据预测结果筛选靠近反应性中心环(RCL)的候选表位,RCL是蛇形蛋白与目标蛋白酶相互作用的区域。针对p1RmS-6和p3RmS-6的抗体可中和RmS-6的活性。使用这种策略,我们能够根据Pepscan结果调整一个计算机算法预测器,以识别另一个serpin中的表位。我们的策略提供了一种具有成本效益的方法来识别蛇形动物中的中和表位。此外,该策略可用于鉴定蛇蛋白和其他蜱虫物种的其他蛋白质的表位,可能导致基于肽的抗蜱虫疫苗的开发。
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引用次数: 0
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
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Molecular and biochemical parasitology
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