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Overexpression of GmCSY3 Enhances Soybean Tolerance to Excess Iron and Aluminum. 过表达GmCSY3增强大豆对过量铁和铝的耐受性
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-05 DOI: 10.3390/biology15010105
Zhuo Liu, Hongqiu Lv, Liying Yang, Yu Wang, Xinqi Zhu, Menghan Chang, Wenwei Liang, Shanshan Wang, Ying Yang, Yining Pan, Changhong Guo, Yingdong Bi, Donglin Guo

Background: Plant citrate synthase (CSY) is involved in the iron deficiency (-Fe) response and aluminum (Al) detoxification. However, knowledge of CSY function in responding to excess iron (+Fe) or Al stress (+Al) is still limited.

Methods: The CDS and promoter of GmCSY3 were isolated from soybean and bioinformatically analyzed. The GmCSY3 expression was detected by qRT-PCR and GUS assay. The growth of GmCSY3 recombinant yeast under +Fe or +Al was detected. The phenotype, CSY activity, citric acid concentration, chlorophyll content, MDA, H2O2, O2- contents, GST, CAT, SOD, and POD activities were examined in GmCSY3 overexpressed and RNAi-suppressed soybean chimeras under +Fe or +Al. Perls and Hematoxylin stained the roots, and the FCR activity was determined.

Results: GmCSY3 was induced by +Fe or +Al, but not by -Fe. GmCSY3 enhanced yeast's acid production and resistance to +Fe or +Al. GmCSY3 overexpression in soybean significantly enhanced CSY activity, promoted growth, alleviated oxidative damage caused by +Fe or +Al, with less free Fe3+ and Al3+, and reduced FCR activity, while GmCSY3 RNAi-suppressed showed the opposite effect.

Conclusions: GmCSY3 promotes the process of citrate synthesis, chelates Fe3+ and Al3+, alleviates oxidative damage caused by +Fe or +Al, and modulates iron absorption in plants.

背景:植物柠檬酸合成酶(CSY)参与缺铁(-Fe)反应和铝(Al)解毒。然而,对于CSY功能对过量铁(+Fe)或铝(+Al)应力的响应的了解仍然有限。方法:从大豆中分离GmCSY3的CDS和启动子,进行生物信息学分析。采用qRT-PCR和GUS法检测GmCSY3的表达。检测GmCSY3重组酵母在+Fe或+Al下的生长情况。研究了+Fe或+Al下GmCSY3过表达和rnai抑制的大豆嵌合体的表型、CSY活性、柠檬酸浓度、叶绿素含量、MDA、H2O2、O2-含量、GST、CAT、SOD和POD活性。紫杉醇和苏木精染色根,测定其FCR活性。结果:+Fe和+Al均能诱导GmCSY3,而-Fe不能。GmCSY3提高了酵母的产酸能力和对+Fe或+Al的抗性。大豆中过表达GmCSY3显著提高CSY活性,促进生长,减轻+Fe或+Al引起的氧化损伤,减少游离Fe3+和Al3+,降低FCR活性,而抑制GmCSY3 rnai则表现出相反的效果。结论:GmCSY3促进柠檬酸盐合成,螯合Fe3+和Al3+,减轻+Fe或+Al引起的氧化损伤,调节植物对铁的吸收。
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引用次数: 0
Molecular Profiling of Foodborne Pathogens in Ready-to-Eat Foods, Al-Madinah Al-Munawarah, Saudi Arabia. 即食食品中食源性病原体的分子分析,Al-Madinah Al-Munawarah,沙特阿拉伯。
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-05 DOI: 10.3390/biology15010104
Omar Almutairi, Ihab M Moussa, Eman Marzouk, Adil Abalkhail, Ayman Elbehiry

Foodborne pathogens remain a global public health concern, and antimicrobial resistance increases their impact. In mass-gathering cities such as Al-Madinah Al-Munawarah, contaminated ready-to-eat (RTE) fast foods can contribute to both local transmission and international spread. In this study, 300 RTE fast food samples, including shawarma, burgers, fried chicken, sandwiches, and salads, were collected from international franchises, local restaurants, and street vendors. Pathogens were identified using conventional culture combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Antimicrobial susceptibility testing followed CLSI guidelines, and real-time PCR confirmed species identity and screened resistance determinants. Principal component analysis (PCA) and dendrogram clustering were used to assess diagnostic discrimination. Among the 300 samples, 129 (43.0%) were culture positive. The most common pathogens were Staphylococcus aureus (14.3%) and Escherichia coli (13.0%), followed by Salmonella spp. (9.0%) and Acinetobacter baumannii (6.7%). About 35% of S. aureus isolates were methicillin resistant (MRSA), and 85% of A. baumannii carried OXA-type carbapenemase genes. MALDI-TOF MS achieved 96.1% score-based identification and, with PCA, showed strong interspecies separation. PCR confirmed species identity and detected widespread resistance genes, with genotype-phenotype concordance of at least 80%. Overall, 60.5% of isolates were multidrug resistant. RTE fast foods in Al-Madinah represent reservoirs of MDR pathogens, including carbapenemase-producing A. baumannii. The combined use of MALDI-TOF MS and real-time PCR established a rapid and scalable workflow that provided reliable identification and resistance profiling in less than 24 h, compared with 48 to 72 h for conventional methods. This approach supports One Health surveillance in high-risk food settings and strengthens preparedness for mass gatherings.

食源性病原体仍然是一个全球公共卫生问题,抗菌素耐药性增加了它们的影响。在麦地那-穆纳瓦拉等人群聚集的城市,受污染的即食快餐既可能导致当地传播,也可能导致国际传播。在本研究中,从国际特许经营,当地餐馆和街头摊贩收集了300种RTE快餐样本,包括沙瓦玛,汉堡,炸鸡,三明治和沙拉。采用常规培养结合基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)对病原菌进行鉴定。抗菌药物敏感性试验遵循CLSI指南,实时PCR证实了物种身份并筛选了耐药决定因素。采用主成分分析(PCA)和树形图聚类评估诊断辨别力。300份样本中培养阳性129份(43.0%)。病原菌以金黄色葡萄球菌(14.3%)和大肠杆菌(13.0%)最多,其次为沙门氏菌(9.0%)和鲍曼不动杆菌(6.7%)。约35%的金黄色葡萄球菌耐甲氧西林(MRSA), 85%的鲍曼不动杆菌携带oxa型碳青霉烯酶基因。MALDI-TOF MS的分值识别率为96.1%,并与主成分分析(PCA)相结合,显示出较强的种间分离。PCR证实了品种的一致性,并检测到广泛存在的抗性基因,基因型-表型一致性至少为80%。总体而言,60.5%的分离株具有多重耐药。麦地那的RTE快餐是耐多药病原体的储藏库,包括产生碳青霉烯酶的鲍曼不动杆菌。MALDI-TOF MS和real-time PCR的联合使用建立了一个快速和可扩展的工作流程,与传统方法的48至72小时相比,在不到24小时内提供可靠的鉴定和耐药性分析。这一方法支持在高风险食品环境中开展“同一个健康”监测,并加强对大规模集会的准备。
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引用次数: 0
Progress in Wildlife Conservation, Management and Biological Research: From Molecular Perspectives to Ecological Processes. 野生动物保护、管理与生物学研究进展:从分子视角到生态过程。
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-05 DOI: 10.3390/biology15010103
Ana Carolina Srbek-Araujo

Wildlife conservation and the management of biological resources face unprecedented challenges in the Anthropocene [...].

野生动物保护和生物资源管理在人类世面临前所未有的挑战[…]。
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引用次数: 0
Chronic Exposure to Niclosamide Disrupts Structure and Metabolism of Digestive Glands and Foot in Cipangopaludina cathayensis. 慢性暴露于氯硝柳胺破坏了中国山核桃消化腺和足的结构和代谢。
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-04 DOI: 10.3390/biology15010102
Yanan Zhang, Yizhen Liu, Qiying Cai, Jun Ye, Tao Wang, Sheng Xu, Gang Ge

Niclosamide has been the primary molluscicide for schistosomiasis control for over 50 years, but its chronic effects on inter-organ interactions in non-target mollusks remain poorly understood. Cipangopaludina cathayensis, a dominant species in East Asian schistosomiasis-endemic regions, was chronically exposed to environmentally relevant concentrations of niclosamide to assess its toxic effects. Digestive glands accumulated more niclosamide than the foot tissues. Prolonged exposure was associated with metabolic impairment of the digestive glands, characterized by tubular atrophy, inflammatory reactions, and depletion of nutrient components. Foot tissues exhibited epithelial lesions and muscle fiber atrophy. Alterations in foot structure were associated with changes in digestive gland nutrient status. Niclosamide exposure may weaken the metabolic coupling between these organs, thereby impairing locomotor function. At the population level, persistent niclosamide exposure may destabilize mollusk trophic-level populations, ultimately leading to ecological consequences. Our findings demonstrate the toxicological risks of niclosamide to freshwater mollusks.

50多年来,氯硝柳胺一直是控制血吸虫病的主要灭虫剂,但其对非靶软体动物器官间相互作用的慢性影响仍知之甚少。东亚血吸虫病流行地区的优势种中国血吸虫(Cipangopaludina cathayensis)长期暴露于环境相关浓度的氯硝柳胺中,以评估其毒性作用。消化腺比足部组织积累了更多的氯硝柳胺。长时间暴露与消化腺的代谢损伤有关,其特征是小管萎缩、炎症反应和营养成分的消耗。足部组织上皮病变,肌纤维萎缩。足部结构的改变与消化腺营养状况的改变有关。氯硝柳胺暴露可能削弱这些器官之间的代谢耦合,从而损害运动功能。在种群水平上,持续接触氯硝柳胺可能破坏软体动物营养水平种群的稳定,最终导致生态后果。我们的研究结果证明了氯硝柳胺对淡水软体动物的毒理学风险。
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引用次数: 0
Enzymatic Antioxidant Defense System of Scots Pine Seedlings Under Conditions of Progressive Manganese Deficiency. 渐进式缺锰条件下松苗的酶促抗氧化防御系统
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-04 DOI: 10.3390/biology15010101
Yury V Ivanov, Alexandra I Ivanova, Alexander V Kartashov, Galina V Glushko, Polina P Loginova, Vladimir V Kuznetsov

Manganese (Mn) is a crucial micronutrient for plants. The impaired function of the oxygen-evolving complex in Photosystem II (PSII) due to Mn deficiency is believed to result in the overproduction of reactive oxygen species and the induction of an enzymatic antioxidant system. In our study, we investigated the effects of progressive Mn deficiency (the difference in Mn content between the needles of control and Mn-deficient plants increased from 17-fold at the beginning of the experiment to 59-fold at the end) on the activities of superoxide dismutase (SOD), catalase, ascorbate peroxidase, and guaiacol peroxidase in the roots and needles of Scots pine seedlings. We found that the soluble protein content in plant organs under Mn deficiency was maintained at a level comparable to that of the control. Regardless of the severity of Mn deficiency, the needles of the Mn-deficient plants presented twofold lower SOD activity than the needles of the control plants. These differences were observed even when Mn deficiency did not negatively affect plant growth. Additionally, the total SOD activity in the needles of both plant groups was determined solely by the activity of the Cu/Zn-containing SOD isozymes. Compared with the control plants, Mn deficiency did not result in an increase in any of the studied H2O2-degrading enzymes in the needles of the seedlings. In contrast, the needles of the Mn-deficient plants presented a lower level of guaiacol peroxidase activity. Despite the inhibition of root growth, Mn deficiency led to changes in the balance of the enzymatic antioxidant system in plant roots. The data obtained suggest that the lack of activation of SOD and other antioxidant enzymes in Scots pine seedlings against the background of progressive Mn deficiency is due to the reduced ability of PSII to generate ROS under these conditions.

锰(Mn)是植物的一种重要微量元素。由于锰缺乏,光系统II (PSII)中进化氧复合物的功能受损被认为会导致活性氧的过量产生和酶促抗氧化系统的诱导。在我们的研究中,我们研究了渐进式缺锰(对照和缺锰植株针叶之间Mn含量的差异从试验开始时的17倍增加到试验结束时的59倍)对松根和针叶中超氧化物歧化酶(SOD)、过氧化氢酶、抗坏血酸过氧化物酶和愈创木酚过氧化物酶活性的影响。结果表明,缺锰处理下植株各器官可溶性蛋白含量维持在与对照相当的水平。无论锰缺乏的严重程度如何,缺锰植株的针叶SOD活性都比对照植株低2倍。这些差异即使在缺锰不影响植物生长的情况下也能观察到。此外,两种植物组的总SOD活性仅由含Cu/ zn的SOD同工酶的活性决定。与对照植株相比,缺锰没有导致幼苗针叶中任何h2o2降解酶的增加。相比之下,缺锰植株的针叶愈创木酚过氧化物酶活性较低。虽然锰缺乏会抑制根系生长,但会导致植物根系酶促抗氧化系统平衡的改变。这些数据表明,在进行性缺锰的背景下,苏格兰松幼苗中SOD和其他抗氧化酶的活性缺乏是由于PSII在这些条件下产生ROS的能力降低。
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引用次数: 0
Transcriptome Sequencing and Differential Analysis of Testes of 1-Year-Old and 3-Year-Old Kazakh Horses. 1岁和3岁哈萨克马睾丸转录组测序和差异分析。
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-04 DOI: 10.3390/biology15010100
Jiahao Liu, Yuting Yang, Liuxiang Wen, Mingyue Wen, Yaqi Zeng, Wanlu Ren, Xinkui Yao

The Kazakh horse is an outstanding dual-purpose dairy and meat breed in China, characterized by early maturity, tolerance to coarse feed, and strong stress resistance. Previous studies have examined gene expression patterns in the testicular tissues of Kazakh horses at different age stages, but the molecular mechanisms regulating testicular sexual maturation remain unclear. To address this gap, this study conducted HE staining and in-depth transcriptome sequencing analysis of Kazakh horse testicular tissue before and after sexual maturity. HE staining showed that the G3 group had well-formed seminiferous tubule lumens, dense interstitial cells, and visible early spermatocytes and spermatozoa, indicating structural maturation. (G1 group: pre-sexual maturity; G3 group: post-sexual maturity), with four biological replicates per group (n = 4). Differentially expressed genes (DEGs) were called using the criteria of |log2(fold change)| ≥ 1.5 and adjusted p-value ≤ 0.05. A total of 3054 differentially expressed genes (DEGs), including CABS1, RPL10, PGAM2, TMSB4X, and CYP17A1, were identified in the G1 and G3 groups. Among these, 402 genes showed upregulation and 2652 genes showed downregulation. GO annotation and KEGG enrichment analysis of DEGs revealed their predominant enrichment in the following categories: signaling pathways such as Focal adhesion, Pathways in cancer, and the PI3K-Akt signaling pathway. RT-qPCR validation confirmed the accuracy of the transcriptomic sequencing data. This study further elucidates the differentially expressed genes and associated signaling pathways in Kazakh stallion testes tissue before and after sexual maturity, providing a theoretical foundation and data reference for enhancing reproductive efficiency in equids and promoting biological processes such as testes development and spermatogenesis.

哈萨克马是中国杰出的奶肉两用品种,具有早熟、耐粗饲料、抗逆性强等特点。以往的研究已经检测了不同年龄阶段哈萨克马睾丸组织中的基因表达模式,但调控睾丸性成熟的分子机制尚不清楚。为了解决这一空白,本研究对哈萨克马性成熟前后的睾丸组织进行了HE染色和深度转录组测序分析。HE染色显示G3组精小管管腔结构完善,间质细胞致密,可见早期精母细胞和精子,结构成熟。(G1组:性成熟前;G3组:性成熟后),每组4个生物重复(n = 4)。以|log2(fold change)|≥1.5、调整后p值≤0.05为标准进行差异表达基因(differential expression genes, DEGs)的命名。在G1和G3组中共鉴定出3054个差异表达基因(DEGs),包括CABS1、RPL10、PGAM2、TMSB4X和CYP17A1。其中402个基因表达上调,2652个基因表达下调。GO注释和对DEGs的KEGG富集分析显示,DEGs主要富集于以下几类:信号通路,如Focal adhesion、癌症通路和PI3K-Akt信号通路。RT-qPCR验证证实了转录组测序数据的准确性。本研究进一步阐明了哈萨克种马性成熟前后睾丸组织的差异表达基因及相关信号通路,为提高马科动物生殖效率,促进睾丸发育、精子发生等生物学过程提供理论基础和数据参考。
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引用次数: 0
Explainable Deep Learning Framework for Reliable Species-Level Classification Within the Genera Desmodesmus and Tetradesmus. 可解释的深度学习框架,在可靠的物种水平分类中,在桥蝇属和四头蝇属。
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-03 DOI: 10.3390/biology15010099
İlknur Meriç Turgut, Dilara Gerdan Koc, Özden Fakıoğlu

Microalgae are an evolutionarily ancient and morphologically diverse group of photosynthetic eukaryotes, with taxonomic resolution complicated by environmentally driven phenotypic plasticity. This study merges deep learning and explainable artificial intelligence (XAI) to establish a transparent, reliable, and biologically meaningful framework for green microalgae (Chlorophyta) classification. Microscope images from three morphologically distinct algal species-Desmodesmus flavescens, Desmodesmus subspicatus, and Tetradesmus dimorphus representing the genera Desmodesmus and Tetradesmus within Chlorophyta-were analyzed using twelve convolutional neural networks, including EfficientNet-B0-B7, DenseNet201, NASNetLarge, Xception, and ResNet152V2. A curated dataset comprising 3624 microscopic images from three Chlorophyta species was used, split into training, validation, and test subsets. All models were trained using standardized preprocessing and data augmentation procedures, including grayscale conversion, CLAHE-based contrast enhancement, rotation, flipping, and brightness normalization. The model's performance was assessed using accuracy and loss metrics on independent test datasets, while interpretability was evaluated through saliency maps and Gradient-weighted Class Activation Mapping (Grad-CAM) visualizations. ResNet152V2 achieved the highest overall performance among all evaluated architectures, outperforming EfficientNet variants, NASNetLarge, and Xception in terms of macro F1-score. Visualization analysis showed that both Grad-CAM and saliency mapping consistently highlighted biologically relevant regions-including cell walls, surface ornamentation, and colony structures-confirming that the models relied on taxonomically meaningful features rather than background artifacts. The findings indicate that the integration of deep learning and XAI can attain consistently high test accuracy for microalgal species, even with constrained datasets. This approach enables automated taxonomy and supports biodiversity monitoring, ecological assessment, biomass optimization, and biodiesel production by integrating interpretability with high predictive accuracy.

微藻是一种进化上古老且形态多样的光合真核生物,其分类分辨率因环境驱动的表型可塑性而变得复杂。本研究将深度学习和可解释人工智能(XAI)相结合,建立了一个透明、可靠、具有生物学意义的绿色微藻分类框架。利用高效网络(EfficientNet-B0-B7)、DenseNet201、NASNetLarge、Xception和ResNet152V2等12个卷积神经网络,对3种形态不同的藻类——黄桥丝藻、次桥丝藻和二形桥丝藻的显微镜图像进行了分析。研究使用了一个精心设计的数据集,其中包括来自三种绿藻物种的3624张显微图像,分为训练、验证和测试子集。所有模型都使用标准化的预处理和数据增强程序进行训练,包括灰度转换、基于clahe的对比度增强、旋转、翻转和亮度归一化。模型的性能通过独立测试数据集的准确性和损失指标进行评估,而可解释性通过显著性图和梯度加权类激活映射(Grad-CAM)可视化来评估。ResNet152V2在所有被评估的体系结构中获得了最高的整体性能,在宏观f1得分方面优于EfficientNet变体、NASNetLarge和Xception。可视化分析表明,Grad-CAM和显著性映射都一致地突出了生物相关区域——包括细胞壁、表面纹饰和菌落结构——证实了模型依赖于分类上有意义的特征,而不是背景人工制品。研究结果表明,即使在有限的数据集下,深度学习和XAI的集成也可以获得一致的高微藻物种测试精度。该方法通过将可解释性与高预测精度相结合,实现了自动化分类,并支持生物多样性监测、生态评估、生物质优化和生物柴油生产。
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引用次数: 0
Characterization of RNA Editing in Oxidative and Glycolytic Skeletal Muscles of Yak. 牦牛氧化和糖酵解骨骼肌RNA编辑特性研究。
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-02 DOI: 10.3390/biology15010097
Yilin Shi, Xuemei Wu, Chunnian Liang, Xian Guo, Xiaoming Ma, Ping Yan, Min Chu, Xiaoyun Wu

Fast-twitch and slow-twitch muscle fibers not only differ in metabolic characteristics and physiological functions but also significantly influence the texture of livestock meat. RNA editing represents an important post-transcriptional regulatory process that can influence both gene expression and the resulting protein function. However, studies on RNA editing events in yak muscle remain limited. This study systematically identified RNA editing events in yak biceps femoris (BF, n = 3) and obliquus externus abdominis (OEA, n = 3) using transcriptomic data, discovering 17,713 unique editing sites, most located in non-coding regions. Within coding regions, 3350 sites were detected, with 1195 resulting in non-synonymous amino acid substitutions. Further analysis revealed that 785 sites potentially affected miRNA binding sites, suggesting RNA editing may participate in miRNA-mediated gene regulation. Tukey's post hoc test (p < 0.05) identified 242 sites (involving 170 genes) with significantly different editing levels between BF and OEA. KEGG pathway analysis indicated that genes with differential RNA editing were predominantly associated with pathways involved in muscle fiber type transitions, including the MAPK and calcium signaling pathways. Collectively, this study maps the RNA editing landscape in yak muscle tissue and identifies distinct, fiber-type-specific RNA editing patterns between oxidative and glycolytic muscle fibers, including differences in editing levels and site distributions, supporting a potential association between RNA editing and muscle fiber type transformation.

家畜快肌纤维和慢肌纤维不仅代谢特性和生理功能不同,而且对肉质质的影响也很大。RNA编辑代表了一个重要的转录后调控过程,可以影响基因表达和由此产生的蛋白质功能。然而,对牦牛肌肉中RNA编辑事件的研究仍然有限。本研究利用转录组学数据系统地鉴定了牦牛股二头肌(BF, n = 3)和腹斜外肌(OEA, n = 3)的RNA编辑事件,发现了17,713个独特的编辑位点,大多数位于非编码区。在编码区中,检测到3350个位点,其中1195个位点导致非同义氨基酸取代。进一步分析发现,785个位点可能影响miRNA结合位点,提示RNA编辑可能参与了miRNA介导的基因调控。Tukey事后检验(p < 0.05)鉴定出BF和OEA之间有242个位点(涉及170个基因)的编辑水平存在显著差异。KEGG通路分析表明,具有差异RNA编辑的基因主要与参与肌纤维类型转换的通路相关,包括MAPK和钙信号通路。总的来说,本研究绘制了牦牛肌肉组织中的RNA编辑景观,并确定了氧化和糖酵解肌纤维之间独特的纤维类型特异性RNA编辑模式,包括编辑水平和位点分布的差异,支持RNA编辑与肌纤维类型转化之间的潜在关联。
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引用次数: 0
Effect of Euterpe oleracea Mart. (Açaí) Seed Bioproducts on Trypanosoma cruzi. 马齿苋的药效。(Açaí)克氏锥虫种子生物制品。
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-02 DOI: 10.3390/biology15010096
Henrique Previtalli-Silva, Daiana de Jesus Hardoim, Raphael de Lucena Banaggia, Carla J Moragas-Tellis, Paulo Victor Ramos de Souza, Maria Dutra Behrens, Thiago de Souza Dias Silva, Kátia da Silva Calabrese, Flávia de Oliveira Cardoso

Chagas disease remains a major public health challenge due to the limited effectiveness and considerable side effects of existing treatments, particularly during the chronic stage. Açaí (Euterpe oleracea) seeds have gained increasing attention as a source of bioactive compounds with potential therapeutic applications. In this study, hydroalcoholic extracts and solvent fractions obtained from açaí seeds were chemically characterized by ESI/MS and HPLC-MS/MS and evaluated for their cytotoxicity and antiparasitic activity against different developmental stages of Trypanosoma cruzi (Y strain). Chemical profiling revealed a predominance of phenolic compounds, particularly catechins and procyanidins, which were identified as major constituents of the hydroalcoholic extract and the ethyl acetate fraction. Cytotoxicity assays performed on murine peritoneal macrophages demonstrated low toxicity, with CC50 values exceeding 500 µg/mL for most samples, indicating a favorable in vitro safety profile. Antiparasitic assays showed weak activity against epimastigote forms; however, significant inhibitory effects were observed against bloodstream trypomastigotes, cell culture-derived trypomastigotes, and intracellular amastigotes. Notably, the hydroalcoholic extract exhibited the highest selectivity against intracellular amastigotes, with a selectivity index greater than 10, fulfilling key criteria proposed by the Drugs for Neglected Diseases initiative (DNDi) for early-stage hit compounds. Flow cytometry analysis showed that both the hydroalcoholic extract and the ethyl acetate fraction induced parasite cell death through late apoptosis-like and necrosis. Together, these findings highlight the antiparasitic potential of E. oleracea seed extracts, particularly against clinically relevant stages of T. cruzi, and support further investigation of these bioproducts as promising candidates for the development of new therapeutic strategies for Chagas disease.

恰加斯病仍然是一项重大的公共卫生挑战,因为现有治疗方法的效力有限,而且副作用很大,特别是在慢性阶段。Açaí (Euterpe oleracea)种子作为一种具有潜在治疗应用的生物活性化合物来源而受到越来越多的关注。本研究采用ESI/MS和HPLC-MS/MS对açaí种子的水醇提取物和溶剂组分进行了化学表征,并对其对克氏锥虫(Y株)不同发育阶段的细胞毒性和抗寄生活性进行了评价。化学分析显示酚类化合物占主导地位,特别是儿茶素和原花青素,它们被确定为水醇提取物和乙酸乙酯部分的主要成分。对小鼠腹膜巨噬细胞进行的细胞毒性试验显示,大多数样品的CC50值超过500µg/mL,表明其具有良好的体外安全性。抗寄生试验显示对附毛鞭毛虫的抗寄生活性较弱;然而,对血流型、细胞培养型和细胞内型无乳线虫有显著的抑制作用。值得注意的是,水酒精提取物对细胞内无梭菌具有最高的选择性,其选择性指数大于10,符合被忽视疾病药物倡议(DNDi)提出的早期命中化合物的关键标准。流式细胞术分析表明,水乙醇提取物和乙酸乙酯部位均通过细胞晚期凋亡样和坏死诱导寄生虫细胞死亡。总之,这些发现突出了甘蓝种子提取物的抗寄生虫潜力,特别是针对临床相关阶段的克氏锥虫,并支持进一步研究这些生物制品作为开发新的恰加斯病治疗策略的有希望的候选物。
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引用次数: 0
Single-Cell Transcriptomic Profiling Reveals That Macrophage-Induced Angiogenesis Contributes to Immunotherapy Resistance in Hepatocellular Carcinoma. 单细胞转录组学分析揭示巨噬细胞诱导的血管生成有助于肝细胞癌的免疫治疗抵抗。
IF 3.5 3区 生物学 Q1 BIOLOGY Pub Date : 2026-01-02 DOI: 10.3390/biology15010095
Xinyu Pan, Baolin Liao, Zhijie Hu, Yuanyan Xiong

Hepatocellular carcinoma (HCC) ranks among the top causes of cancer-related mortality worldwide, and its complex tumor microenvironment (TME) contributes to poor responses to immunotherapy. Although PD-1/PD-L1 blockade has emerged as an effective treatment strategy, therapeutic resistance frequently limits its clinical benefit. Here, we uncover a distinct macrophage population associated with anti-PD-1 resistance in HCC. Single-cell transcriptomic profiling reveals an NFKBIZ+ M0 macrophage subset predominantly present in non-responders. Notably, these macrophages exhibit a hypoxia-induced phenotype characterized by the secretion of VEGFA and HBEGF, which cooperatively enhance tumor angiogenesis, alongside an elevated expression of the inflammatory chemokines CXCL2, CXCL3, and CXCL8 that consolidate an immunosuppressive, pro-tumorigenic microenvironment. Transcriptional regulatory network analysis further identified FOSB-VEGFA and FOS-HBEGF axes as key drivers of this pathogenic macrophage phenotype. Our findings define a distinct NFKBIZ+ macrophage population that mechanistically links hypoxia, angiogenesis, and immune evasion to PD-1 blockade resistance. This work provides new insights into the cellular and molecular basis of immunotherapy failure in HCC and highlights potential targets for overcoming treatment resistance.

肝细胞癌(HCC)是全球癌症相关死亡的主要原因之一,其复杂的肿瘤微环境(TME)导致免疫治疗反应不佳。虽然PD-1/PD-L1阻断已成为一种有效的治疗策略,但治疗耐药性往往限制了其临床效益。在这里,我们发现了一种与HCC中抗pd -1耐药性相关的独特巨噬细胞群。单细胞转录组学分析显示NFKBIZ+ M0巨噬细胞亚群主要存在于无应答者中。值得注意的是,这些巨噬细胞表现出缺氧诱导的表型,其特征是VEGFA和HBEGF的分泌,它们协同促进肿瘤血管生成,同时炎症趋化因子CXCL2、CXCL3和CXCL8的表达升高,这些趋化因子巩固了免疫抑制、促肿瘤发生的微环境。转录调控网络分析进一步发现FOSB-VEGFA和FOS-HBEGF轴是这种致病性巨噬细胞表型的关键驱动因素。我们的研究结果定义了一个独特的NFKBIZ+巨噬细胞群,它将缺氧、血管生成和免疫逃避与PD-1阻断抵抗机制联系起来。这项工作为HCC免疫治疗失败的细胞和分子基础提供了新的见解,并强调了克服治疗耐药的潜在靶点。
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Biology-Basel
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