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Real-world use of imipenem/cilastatin/relebactam for the treatment of KPC-producing Klebsiella pneumoniae complex and difficult-to-treat resistance (DTR) Pseudomonas aeruginosa infections: a single-center preliminary experience 亚胺培南/西司他丁/雷巴坦在治疗产KPC肺炎克雷伯菌复合菌和难治性(DTR)铜绿假单胞菌感染中的实际应用:单中心初步经验
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1432296
Cristiana Leanza, M. Mascellino, L. Volpicelli, S. Covino, Antonio Falletta, F. Cancelli, Cristiana Franchi, Martina Carnevalini, C. Mastroianni, Alessandra Oliva
Real-life experience with imipenem/cilastatin/relebactam (IMI/REL) for the treatment of KPC-producing Klebsiella pneumoniae complex (KPC-Kp) and difficult-to-treat resistance (DTR) Pseudomonas aeruginosa (DTR-PA) infections is herein described.Adult patients with KPC-Kp or DTR-PA infections who received ≥48 h of IMI/REL were included. Clinical and microbiological outcomes were retrieved through the medical records. Primary outcome was clinical cure. Secondary outcomes included mortality from infection onset and adverse effects attributable to IMI/REL.We included 10 patients with different infections caused by DTR-PA (n = 4), KPC-Kp [n = 5, of which 3 ceftazidime/avibactam-resistant (CTV-R KPC-Kp), 2 CTV susceptible (CTV-S KPC-Kp)] or both DTR-PA/KPC-Kp (n = 1) successfully treated with IMI/REL: 3 hospital-acquired pneumonia, 1 ventilator-associated pneumonia, 2 skin and soft tissue infections, 1 osteomyelitis, 2 bloodstream infections, 1 complicated urinary tract infection. Clinical cure was achieved in all cases. No patients died and no side effect were reported.We reported the preliminary real-life experience on the successful and safe use of IMI/REL for the treatment of KPC-Kp or DTR-PA complicated infections, including pneumonia and bone infections.
本文介绍了亚胺培南/西司他丁/雷巴坦(IMI/REL)治疗产KPC肺炎克雷伯菌复合体(KPC-Kp)和难治耐药(DTR)铜绿假单胞菌(DTR-PA)感染的实际经验。通过病历检索临床和微生物学结果。主要结果为临床治愈。我们纳入了 10 例由 DTR-PA(n = 4)、KPC-Kp(n = 5,其中 3 例头孢他啶/阿维菌素耐药(CTV-R KPC-Kp),2 例 CTV 易感(CTV-S KPC-Kp))或 DTR-PA/KPC-Kp 同时感染(n = 1)引起的不同感染患者,他们均成功接受了 IMI/REL:3 例医院获得性肺炎、1 例呼吸机相关肺炎、2 例皮肤和软组织感染、1 例骨髓炎、2 例血流感染、1 例复杂性尿路感染。所有病例均临床治愈。我们报告了成功安全使用 IMI/REL 治疗 KPC-Kp 或 DTR-PA 并发感染(包括肺炎和骨感染)的初步实际经验。
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引用次数: 0
Age-correlated changes in the canine oral microbiome 犬类口腔微生物群与年龄相关的变化
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1426691
Gregory Kislik, Lin Zhou, L. Rubbi, Matteo Pellegrini
Canine oral disease has been associated with significant changes in the oral microbiome rather than the presence or absence of individual species. In addition, most studies focus on a single age group of canines and as of yet, the relationship between canine microbiomes and age is poorly understood.This study used a shotgun whole gene sequencing approach in tandem with the Aladdin Bioinformatics platform to profile the microbiomes of 96 companion dogs, with the sourmash-zymo reference database being used to perform taxonomic profiling.Findings showed significant age correlations among 19 species, including positive correlations among several Porphyromonas species and a negative correlation with C. steedae. Although a significant correlation was found between predicted and actual ages, ElasticNet Regression was unable to successfully predict the ages of younger canines based on their microbiome composition. Both microbiome samples and microbial species were successfully clustered by age group or age correlation, showing that the age-microbiome relationship survives dimensionality reduction. Three distinct clusters of microbial species were found, which were characterized by Porphyromonas, Conchiformibius, and Prevotella genera, respectively.Findings showed that the microbiomes of older dogs resembled those that previous literature attributed to dogs with periodontal disease. This suggests that the process of aging may introduce greater risks for canine oral disease.
犬口腔疾病与口腔微生物组的显著变化有关,而不是与个别物种的存在或缺乏有关。本研究使用霰弹枪全基因测序方法和阿拉丁生物信息学平台对 96 只伴侣犬的微生物组进行分析,并使用 sourmash-zymo 参考数据库进行分类分析。虽然预测年龄与实际年龄之间存在明显的相关性,但根据微生物组的组成,ElasticNet 回归法无法成功预测较年轻犬科动物的年龄。微生物组样本和微生物物种都成功地按年龄组或年龄相关性进行了聚类,这表明年龄与微生物组之间的关系经得起降维处理。研究结果表明,老年犬的微生物组与以往文献中认为患有牙周病的犬的微生物组相似。这表明,衰老过程可能会给犬的口腔疾病带来更大的风险。
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引用次数: 0
BlpU is a broad-spectrum bacteriocin in Streptococcus thermophilus BlpU 是嗜热链球菌中的一种广谱细菌素
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1409359
J. Renye, G. A. Somkuti, Phoebe X. Qi, D. H. Steinberg, Michael J. McAnulty, Amanda L. Miller, G. Guron, Adam M. Oest
Streptococcus thermophilus strain B59671 naturally produces thermophilin 110, a broad-spectrum bacteriocin encoded within the bacteriocin-like peptide (blp) gene cluster, and thermophilin 13 from a separate chromosomal locus. Analysis of the blp gene cluster revealed two genes, blpU and blpK, as potentially encoding bacteriocins. Deletion of blpK from the B59671 chromosome did not result in a loss of antimicrobial activity against either S. thermophilus ST113 or Pediococcus acidilactici F. A deletion mutant of blpU could not be generated in B59671, but the mature BlpU peptide obtained through overexpression in E. coli BL21 or chemical synthesis inhibited the growth of S. thermophilus strains, Streptococcus mutans UA159, P. acidilactici F, and Listeria innocua GV9 L-S, evidencing as a broad-spectrum bacteriocin that does not require modification for activity. This study also showed that the transcription of blpU was approximately 16-fold higher in B59671 than in an induced culture of S. thermophilus LMD-9, which produces a blp-encoded bacteriocin. The increased expression of BlpU in B59671 may explain the unique antimicrobial spectrum associated with this strain. Additionally, it was shown that a blpC deletion mutant of B59671, which prevents expression of BlpU and BlpK, inhibited the growth of other S. thermophilus strains and Bacillus cereus, suggesting that thermophilin 13 produced by B59671 possessed both intra- and interspecies antimicrobial activity. While this study confirmed that BlpU can function as an independent antimicrobial peptide, further studies are required to determine if BlpK can function independently as a broad-spectrum antimicrobial.
嗜热链球菌(Streptococcus thermophilus)菌株 B59671 可天然产生嗜热菌素 110(一种广谱细菌素,在类细菌素肽(blp)基因簇中编码)和嗜热菌素 13(来自一个单独的染色体位点)。对 blp 基因簇的分析表明,blpU 和 blpK 这两个基因可能编码细菌素。从 B59671 染色体中缺失 blpK 不会导致对嗜热菌 ST113 或酸性球菌 F 的抗菌活性丧失。在 B59671 中无法产生 blpU 的缺失突变体,但通过在大肠杆菌 BL21 中过表达或化学合成获得的成熟 BlpU 肽可抑制嗜热菌菌株、变异链球菌 UA159、酸性球菌 F 和无毒李斯特菌 GV9 L-S 的生长,证明它是一种广谱细菌素,不需要修饰就具有活性。这项研究还表明,B59671 中 blpU 的转录量比嗜热菌 LMD-9 的诱导培养物中高出约 16 倍,而嗜热菌 LMD-9 能产生由 blp 编码的细菌素。B59671 中 BlpU 表达量的增加可能是该菌株具有独特抗菌谱的原因。此外,研究还表明,B59671的blpC缺失突变体阻止了BlpU和BlpK的表达,抑制了其他嗜热菌菌株和蜡样芽孢杆菌的生长,这表明B59671产生的嗜热菌素13具有种内和种间抗菌活性。虽然这项研究证实了 BlpU 可以作为一种独立的抗菌肽发挥作用,但要确定 BlpK 是否可以作为一种广谱抗菌素独立发挥作用,还需要进一步的研究。
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引用次数: 0
The functional identification and evaluation of endophytic bacteria sourced from the roots of tolerant Achyranthes bidentata to overcome monoculture problems of Rehmannia glutinosa 从耐受性牛膝根中提取内生细菌的功能鉴定和评估,以克服地黄的单一栽培问题
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1399406
Chunli Zeng, Yazhou Liu, Bianhong Zhang, Chenjing Zhang, Niu Li, Leshan Ji, Chaojie Lan, Bin Qin, Yuncheng Yang, Juanying Wang, Ting Chen, Changxun Fang, Wenxiong Lin
The isolation and identification of plant growth-promoting endophytic bacteria (PGPEB) from Achyranthes bidentata roots have profound theoretical and practical implications in ecological agriculture, particularly as bio-inoculants to address challenges associated with continuous monoculture. Our research revealed a significant increase in the abundance of these beneficial bacteria in A. bidentata rhizosphere soil under prolonged monoculture conditions, as shown by bioinformatics analysis. Subsequently, we isolated 563 strains of endophytic bacteria from A. bidentata roots. Functional characterization highlighted diverse plant growth-promoting traits among these bacteria, including the secretion of indole-3-acetic acid (IAA) ranging from 68.01 to 73.25 mg/L, phosphorus and potassium solubilization capacities, and antagonistic activity against pathogenic fungi (21.54%−50.81%). Through 16S rDNA sequencing, we identified nine strains exhibiting biocontrol and growth-promoting potential. Introduction of a synthetic microbial consortium (SMC) in pot experiments significantly increased root biomass by 48.19% in A. bidentata and 27.01% in replanted Rehmannia glutinosa. These findings provide innovative insights and strategies for addressing continuous cropping challenges, highlighting the practical promise of PGPEB from A. bidentata in ecological agriculture to overcome replanting obstacles for non-host plants like R. glutinosa, thereby promoting robust growth in medicinal plants.
从牛膝(Achyranthes bidentata)根部分离和鉴定植物生长促进内生细菌(PGPEB)对生态农业具有深远的理论和实践意义,尤其是作为生物接种剂来应对连续单一栽培带来的挑战。通过生物信息学分析,我们的研究发现,在长期单一栽培条件下,这些有益细菌在牛膝根圈土壤中的丰度明显增加。随后,我们从双戟根中分离出了 563 株内生细菌。功能表征显示,这些细菌具有多种促进植物生长的特性,包括分泌 68.01 至 73.25 mg/L 的吲哚-3-乙酸(IAA)、溶解磷和钾的能力以及对病原真菌的拮抗活性(21.54%-50.81%)。通过 16S rDNA 测序,我们确定了 9 株具有生物防治和生长促进潜力的菌株。在盆栽实验中引入合成微生物菌群(SMC),可显著增加双叶地黄的根部生物量 48.19%,以及移栽地黄的根部生物量 27.01%。这些发现为应对连作挑战提供了创新见解和策略,凸显了二叉地黄丸的 PGPEB 在生态农业中克服谷氨酰胺等非寄主植物移栽障碍的实用前景,从而促进药用植物的茁壮成长。
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引用次数: 0
Respiratory diseases and gut microbiota: relevance, pathogenesis, and treatment 呼吸系统疾病与肠道微生物群:相关性、发病机制和治疗
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1358597
Mengdi Sun, Fang Lu, Donghua Yu, Yu Wang, Pingping Chen, Shumin Liu
Preclinical evidence has firmly established a bidirectional interaction among the lung, gut, and gut microbiome. There are many complex communication pathways between the lung and intestine, which affect each other's balance. Some metabolites produced by intestinal microorganisms, intestinal immune cells, and immune factors enter lung tissue through blood circulation and participate in lung immune function. Altered gut–lung–microbiome interactions have been identified in rodent models and humans of several lung diseases such as pulmonary fibrosis, chronic obstructive pulmonary disease, lung cancer, asthma, etc. Emerging evidence suggests that microbial therapies can prevent and treat respiratory diseases, but it is unclear whether this association is a simple correlation with the pathological mechanisms of the disease or the result of causation. In this review, we summarize the complex and critical link between the gut microbiota and the lung, as well as the influence and mechanism of the gut microbiota on respiratory diseases, and discuss the role of interventions such as prebiotics and fecal bacteria transplantation on respiratory diseases. To provide a reference for the rational design of large-scale clinical studies, the direct application of microbial therapy to respiratory-related diseases can reduce the incidence and severity of diseases and accompanying complications.
临床前证据已牢固确立了肺、肠道和肠道微生物群之间的双向互动关系。肺和肠道之间有许多复杂的交流途径,它们会影响彼此的平衡。肠道微生物、肠道免疫细胞和免疫因子产生的一些代谢产物通过血液循环进入肺组织,参与肺免疫功能。在肺纤维化、慢性阻塞性肺病、肺癌、哮喘等多种肺部疾病的啮齿类动物模型和人体中,已发现肠道-肺-微生物组之间的相互作用发生了改变。新的证据表明,微生物疗法可以预防和治疗呼吸系统疾病,但目前还不清楚这种关联是与疾病的病理机制简单相关还是因果关系的结果。在这篇综述中,我们总结了肠道微生物群与肺之间复杂而关键的联系,以及肠道微生物群对呼吸系统疾病的影响和机制,并讨论了益生元和粪菌移植等干预措施对呼吸系统疾病的作用。为合理设计大规模临床研究提供参考,将微生物疗法直接应用于呼吸系统相关疾病,可降低疾病的发病率和严重程度以及伴随的并发症。
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引用次数: 0
Robust anti-tubercular profile of Solanum virginianum extract in enhancing isoniazid bioavailability and curtailing stress tolerance in Mycobacterium smegmatis 茄属植物提取物在提高异烟肼生物利用度和降低分枝杆菌应激耐受性方面具有强大的抗结核作用
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1429027
A. Balkrishna, M. Joshi, Manisha Kabdwal, Meenu Tomer, Savita Lochab, Anurag Varshney
The formidable survival mechanisms employed by Mycobacterium tuberculosis (Mtb), combined with the low bioavailability of anti-tubercular drugs and their associated hepatotoxicity, worsen tuberculosis management. Traditional medicinal plants offer potential solutions to these challenges. This study focuses on exploring the anti-tubercular potential of Solanum virginianum against Mycobacterium smegmatis, mc2155.HPTLC and UHPLC phytochemically characterized the hydro-methanolic extract of Solanum virginianum (SVE). SVE curtails the growth and viability of mc2155 under normal and in vitro stress conditions. The compromised cell wall integrity of mc2155 with SVE is depicted through scanning electron microscopy (SEM) while EtBr permeability assays and TLC-based comparative changes in lipids extraction addressed the integrity of the cell wall. Furthermore, SVE augmented the susceptibility of mc2155 towards Isoniazid (INH) through enhanced bioavailability. Adjunct treatment of SVE with INH demonstrated a markedly reduced survival of the intracellular bacilli. The study also uncovered the hepatoprotective potential of SVE in HepG2 cells.This research paves the way for deeper exploration into the potential of Solanum virginianum against virulent Mtb strains, emphasizing over the significance of traditional medicinal plants in tuberculosis treatment. Collectively, the findings suggest SVE as a potent candidate for independent or adjunct anti-tubercular therapy.
结核分枝杆菌(Mtb)具有强大的生存机制,加上抗结核药物的生物利用度低及其相关的肝脏毒性,使得结核病的治疗更加困难。传统药用植物为应对这些挑战提供了潜在的解决方案。这项研究的重点是探索茄属植物对烟草分枝杆菌 mc2155 的抗结核潜力。HPTLC 和 UHPLC 植物化学鉴定了茄属植物水甲醇提取物(SVE)。在正常和体外应激条件下,SVE 可抑制 mc2155 的生长和存活。扫描电子显微镜(SEM)显示了SVE对mc2155细胞壁完整性的损害,而EtBr渗透性测定和基于TLC的脂质提取比较变化则反映了细胞壁的完整性。此外,SVE 通过提高生物利用度增强了 mc2155 对异烟肼(INH)的敏感性。用 INH 对 SVE 进行辅助治疗后,细胞内杆菌的存活率明显降低。这项研究为深入探讨茄属植物抗毒性 Mtb 菌株的潜力铺平了道路,强调了传统药用植物在结核病治疗中的重要意义。总之,研究结果表明茄属植物是独立或辅助抗结核治疗的有效候选药物。
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引用次数: 0
The infants’ gut microbiome: setting the stage for the early onset of obesity 婴儿肠道微生物群:为肥胖症的早期发病埋下伏笔
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1371292
Yvonne Vallès, Muhammad Arshad, Mamoun Abdalbaqi, Claire K. Inman, Amar Ahmad, Nizar Drou, Kristin C. Gunsalus, Raghib Ali, Muna Tahlak, A. Abdulle
In the past three decades, dietary and lifestyle changes worldwide have resulted in a global increase in the prevalence of obesity in both adults and children. Known to be highly influenced by genetic, environmental and lifestyle factors, obesity is characterized by a low-grade chronic inflammation that contributes to the development of other metabolic diseases such as diabetes and cardiovascular disease. Recently, the gut microbiome has been added as a cause/contributor to the development of obesity. As differences in the microbiome between obese and normoweight individuals have been observed, we set out to determine whether infants harbor an obesogenic microbiome early on and whether the pre-pregnancy status of the mother (obese or normoweight) is correlated to their infant’s microbiome composition. Using shotgun sequencing, we analyzed stool samples throughout the first year of life from infants born to obese (n = 23 participants, m = 104 samples) and normoweight (n = 23 participants, m = 99 samples) mothers. We found that the infants’ microbiome diversity at taxonomic and functional levels was significantly influenced by time (ANOVA p < 0.001) but not by the mother’s pre-pregnancy status. Overall, no deterministic succession of taxa or functions was observed. However, infants born to obese mothers were found to have a significantly higher Bacillota/Bacteroidota ratio (p = 0.02) at six months, were significantly depleted from six months old of the well-established obesity biomarkers Akkermansia municiphila and Faecalibacterium prausnitzii (p < 0.01), and were at one week old, significantly enriched in pathways such as the UDP-N-acetyl-D-glucosamine biosynthesis II (p = 0.02) involved in leptin production, suggesting perhaps that there may exist some underlying mechanisms that dictate the development of an obesogenic microbiota early on.
在过去的三十年里,全球范围内饮食和生活方式的改变导致了全球成人和儿童肥胖症发病率的上升。众所周知,肥胖症受遗传、环境和生活方式等因素的影响很大,其特点是低度慢性炎症,会导致糖尿病和心血管疾病等其他代谢性疾病的发生。最近,肠道微生物组也被认为是肥胖的诱因之一。由于已观察到肥胖者和正常体重者的微生物组存在差异,我们着手确定婴儿是否在早期就携带致肥胖微生物组,以及母亲的孕前状态(肥胖或正常体重)是否与婴儿的微生物组组成相关。我们使用霰弹枪测序技术分析了肥胖母亲(23 人,104 个样本)和正常体重母亲(23 人,99 个样本)所生婴儿在出生后第一年内的粪便样本。我们发现,婴儿微生物组在分类学和功能水平上的多样性受到时间的显著影响(方差分析 p < 0.001),但不受母亲孕前状态的影响。总体而言,没有观察到分类群或功能的确定性继承。然而,研究发现,肥胖母亲所生的婴儿在六个月大时,杆菌群/类杆菌群比率明显较高(p = 0.02),六个月大时,肥胖生物标志物 Akkermansia municiphila 和 Faecalibacterium prausnitzii 明显减少(p < 0.01),而在一周大时,参与瘦素生成的 UDP-N- 乙酰-D-葡萄糖胺生物合成 II(p = 0.02)等通路明显富集,这表明可能存在某些潜在机制,决定了致肥胖微生物群的早期发展。
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引用次数: 0
Mechanistic insights into rumen function promotion through yeast culture (Saccharomyces cerevisiae) metabolites using in vitro and in vivo models 利用体外和体内模型深入了解酵母培养物(酿酒酵母)代谢物促进瘤胃功能的机理
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1407024
Xue Chen, Jun Xiao, Wanzhu Zhao, Yanan Li, Wei Zhao, Weigang Zhang, Liang Xin, Zhiyi Han, Lanhui Wang, N. D. Aschalew, Xuefeng Zhang, Tao Wang, Guixin Qin, Zhe Sun, Y. Zhen
Yeast culture (YC) enhances ruminant performance, but its functional mechanism remains unclear because of the complex composition of YC and the uncertain substances affecting rumen fermentation. The objective of this study was to determine the composition of effective metabolites in YC by exploring its effects on rumen fermentation in vitro, growth and slaughter performance, serum index, rumen fermentation parameters, rumen microorganisms, and metabolites in lambs.In Trial 1, various YCs were successfully produced, providing raw materials for identifying effective metabolites. The experiment was divided into 5 treatment groups with 5 replicates in each group: the control group (basal diet without additives) and YC groups were supplemented with 0.625‰ of four different yeast cultures, respectively (groups A, B, C, and D). Rumen fermentation parameters were determined at 3, 6, 12, and 24 h in vitro. A univariate regression model multiple factor associative effects index (MFAEI; y) was established to correlate the most influential factors on in vitro rumen fermentation with YC metabolites (x). This identified the metabolites promoting rumen fermentation and optimal YC substance levels. In Trial 2, metabolites in YC not positively correlated with MFAEI were excluded, and effective substances were combined with pure chemicals (M group). This experiment validated the effectiveness of YC metabolites in lamb production based on their impact on growth, slaughter performance, serum indices, rumen parameters, microorganisms, and metabolites. Thirty cross-generation rams (Small tail Han-yang ♀ × Australian white sheep ♂) with good body condition and similar body weight were divided into three treatment groups with 10 replicates in each group: control group, YC group, pure chemicals combination group (M group).Growth performance and serum index were measured on days 30 and 60, and slaughter performance, rumen fermentation parameters, microorganisms, and metabolites were measured on day 60. The M group significantly increased the dressing percentage, and significantly decreased the GR values of lambs (p  < 0.05). The concentration of growth hormone (GH), Cortisol, insulin (INS), and rumen VFA in the M group significantly increased (p < 0.05).These experiments confirmed that YC or its screened effective metabolites positively impact lamb slaughter performance, rumen fermentation, and microbial metabolism.
酵母培养物(YC)可提高反刍动物的生产性能,但由于 YC 成分复杂,影响瘤胃发酵的物质不确定,因此其功能机制仍不清楚。本研究旨在通过探讨酵母培养物对羔羊体外瘤胃发酵、生长和屠宰性能、血清指数、瘤胃发酵参数、瘤胃微生物和代谢物的影响,确定酵母培养物中有效代谢物的组成。试验分为 5 个处理组,每组 5 个重复:对照组(不含添加剂的基础日粮)和分别添加 0.625‰四种不同酵母培养物的 YC 组(A、B、C 和 D 组)。在体外 3、6、12 和 24 小时测定瘤胃发酵参数。建立了单变量回归模型多因素关联效应指数(MFAEI;y),将对体外瘤胃发酵影响最大的因素与 YC 代谢物(x)联系起来。这就确定了促进瘤胃发酵的代谢物和最佳 YC 物质水平。在试验 2 中,剔除了 YC 中与 MFAEI 不呈正相关的代谢物,并将有效物质与纯化学物质结合(M 组)。该试验根据 YC 代谢物对羔羊生长、屠宰性能、血清指数、瘤胃参数、微生物和代谢物的影响,验证了 YC 代谢物在羔羊生产中的有效性。将 30 只体况良好、体重相近的杂交一代公羊(小尾寒羊♀×澳洲白羊♂)分为三个处理组,每组 10 个重复:对照组、YC 组、纯药组合组(M 组)。结果表明,M 组明显提高了羔羊的换料率,明显降低了 GR 值(P < 0.05)。这些实验证实,YC 或其筛选出的有效代谢物对羔羊的屠宰性能、瘤胃发酵和微生物代谢有积极影响。
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引用次数: 0
Trypanosoma brucei multi-aminoacyl-tRNA synthetase complex formation limits promiscuous tRNA proofreading 布氏锥虫多氨基酸-tRNA合成酶复合物的形成限制了杂乱的tRNA校对工作
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1445687
Rylan R. Watkins, Anna Vradi, Irina Shulgina, Karin Musier-Forsyth
Faithful mRNA decoding depends on the accuracy of aminoacyl-tRNA synthetases (ARSs). Aminoacyl-tRNA proofreading mechanisms have been well-described in bacteria, humans, and plants. However, our knowledge of translational fidelity in protozoans is limited. Trypanosoma brucei (Tb) is a eukaryotic, protozoan pathogen that causes Human African Trypanosomiasis, a fatal disease if untreated. Tb undergoes many physiological changes that are dictated by nutrient availability throughout its insect-mammal lifecycle. In the glucose-deprived insect vector, the tsetse fly, Tb use proline to make ATP via mitochondrial respiration. Alanine is one of the major by-products of proline consumption. We hypothesize that the elevated alanine pool challenges Tb prolyl-tRNA synthetase (ProRS), an ARS known to misactivate alanine in all three domains of life, resulting in high levels of misaminoacylated Ala-tRNAPro. Tb encodes two domains that are members of the INS superfamily of aminoacyl-tRNA deacylases. One homolog is appended to the N-terminus of Tb ProRS, and a second is the major domain of multi-aminoacyl-tRNA synthetase complex (MSC)-associated protein 3 (MCP3). Both ProRS and MCP3 are housed in the Tb MSC. Here, we purified Tb ProRS and MCP3 and observed robust Ala-tRNAPro deacylation activity from both enzymes in vitro. Size-exclusion chromatography multi-angle light scattering used to probe the oligomerization state of MCP3 revealed that although its unique N-terminal extension confers homodimerization in the absence of tRNA, the protein binds to tRNA as a monomer. Kinetic assays showed MCP3 alone has relaxed tRNA specificity and promiscuously hydrolyzes cognate Ala-tRNAAla; this activity is significantly reduced in the presence of Tb alanyl-tRNA synthetase, also housed in the MSC. Taken together, our results provide insight into translational fidelity mechanisms in Tb and lay the foundation for exploring MSC-associated proteins as novel drug targets.
可靠的 mRNA 解码取决于氨基酰-tRNA 合成酶(ARS)的准确性。氨基酰-tRNA 校对机制在细菌、人类和植物中都有详细描述。然而,我们对原生动物的翻译保真度了解有限。布氏锥虫(Tb)是一种真核原生动物病原体,可引起人类非洲锥虫病,如果不及时治疗,将导致死亡。Tb 在整个昆虫-哺乳动物生命周期中会发生许多生理变化,这些变化取决于营养物质的供应情况。在缺乏葡萄糖的昆虫载体采采蝇中,Tb 通过线粒体呼吸利用脯氨酸制造 ATP。丙氨酸是消耗脯氨酸的主要副产品之一。我们推测,丙氨酸池的升高对 Tb 丙氨酰-tRNA 合成酶(ProRS)提出了挑战,已知这种 ARS 在生命的所有三个领域中都会误动作丙氨酸,从而导致高水平的误氨基化 Ala-tRNAPro。Tb 编码两个属于 INS 超家族氨基酰-tRNA 脱酰酶的结构域。其中一个同源物附加在 Tb ProRS 的 N 端,第二个同源物是多氨基酰-tRNA 合成酶复合体(MSC)相关蛋白 3(MCP3)的主要结构域。ProRS 和 MCP3 都位于 Tb MSC 中。在这里,我们纯化了 Tb ProRS 和 MCP3,并在体外观察到这两种酶具有强大的 Ala-tRNAPro 脱酰化活性。用尺寸排阻色谱多角度光散射法探测 MCP3 的寡聚状态发现,虽然其独特的 N 端延伸在没有 tRNA 的情况下具有同源二聚体化的特性,但该蛋白是以单体形式与 tRNA 结合的。动力学分析表明,MCP3本身具有松弛的tRNA特异性,并能杂乱地水解同源的Ala-tRNAAla;如果有同样位于间充质干细胞中的Tb丙氨酰-tRNA合成酶存在,这种活性就会显著降低。综上所述,我们的研究结果提供了对 Tb 翻译保真度机制的深入了解,并为探索 MSC 相关蛋白作为新型药物靶标奠定了基础。
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引用次数: 0
Unveiling metabolic pathways of selected plant-derived glycans by Bifidobacterium pseudocatenulatum 揭示假双歧杆菌代谢某些植物源聚糖的途径
Pub Date : 2024-07-16 DOI: 10.3389/fmicb.2024.1414471
Rocio Sanchez-Gallardo, Francesca Bottacini, Lisa Friess, M. Esteban-Torres, Clarissa Somers, Rebecca L. Moore, F. McAuliffe, Paul D. Cotter, D. van Sinderen
Bifidobacteria are commonly encountered members of the human gut microbiota that possess the enzymatic machinery necessary for the metabolism of certain plant-derived, complex carbohydrates. In the current study we describe differential growth profiles elicited by a panel of 21 newly isolated Bifidobacterium pseudocatenulatum strains on various plant-derived glycans. Using a combination of gene-trait matching and comparative genome analysis, we identified two distinct xylanases responsible for the degradation of xylan. Furthermore, three distinct extracellular α-amylases were shown to be involved in starch degradation by certain strains of B. pseudocatenulatum. Biochemical characterization showed that all three α-amylases can cleave the related substrates amylose, amylopectin, maltodextrin, glycogen and starch. The genes encoding these enzymes are variably found in the species B. pseudocatenulatum, therefore constituting a strain-specific adaptation to the gut environment as these glycans constitute common plant-derived carbohydrates present in the human diet. Overall, our study provides insights into the metabolism of these common dietary carbohydrates by a human-derived bifidobacterial species.
双歧杆菌是人类肠道微生物群中常见的成员,它们拥有代谢某些植物源复杂碳水化合物所需的酶机制。在目前的研究中,我们描述了 21 株新分离的双歧杆菌在各种植物衍生糖类上引起的不同生长特征。通过基因性状匹配和比较基因组分析,我们发现了两种不同的木聚糖酶,它们负责降解木聚糖。此外,三种不同的胞外α-淀粉酶被证明参与了假淀粉芽孢杆菌某些菌株的淀粉降解。生化鉴定表明,这三种α-淀粉酶都能分解相关的底物淀粉、直链淀粉、麦芽糊精、糖原和淀粉。编码这些酶的基因在假中华酵母菌中存在差异,因此构成了菌株对肠道环境的特异性适应,因为这些糖构成了人类饮食中常见的植物衍生碳水化合物。总之,我们的研究为了解源自人类的双歧杆菌物种对这些常见膳食碳水化合物的新陈代谢提供了见解。
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Frontiers in Microbiology
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