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Nitric oxide mediates positive regulation of Nostoc flagelliforme polysaccharide yield via potential S-nitrosylation of G6PDH and UGDH. 一氧化氮通过对 G6PDH 和 UGDH 潜在的 S-亚硝基化作用,介导对 Nostoc flagelliforme 多糖产量的正向调节。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-22 DOI: 10.1186/s12896-024-00884-z
Meng-Yuan Li, Yan-Ru Li, Cheng-Feng Han, Jie Zhang, Rui-Ying Zhu, Yan Zhang, Jian Li, Shi-Ru Jia, Pei-Pei Han

Based on our previous findings that salicylic acid and jasmonic acid increased Nostoc flagelliforme polysaccharide yield by regulating intracellular nitric oxide (NO) levels, the mechanism through which NO affects polysaccharide biosynthesis in Nostoc flagelliforme was explored from the perspective of S-nitrosylation (SNO). The addition of NO donor and scavenger showed that intracellular NO had a significant positive effect on the polysaccharide yield of N. flagelliforme. To explore the mechanism, we investigated the relationship between NO levels and the activity of several key enzymes involved in polysaccharide biosynthesis, including fructose 1,6-bisphosphate aldolase (FBA), glucokinase (GK), glucose 6-phosphate dehydrogenase (G6PDH), mitochondrial isocitrate dehydrogenase (ICDH), and UDP-glucose dehydrogenase (UGDH). The enzymatic activities of G6PDH, ICDH, and UGDH were shown to be significantly correlated with the shifts in intracellular NO levels. For further validation, G6PDH, ICDH, and UGDH were heterologously expressed in Escherichia coli and purified via Ni+-NAT affinity chromatography, and subjected to a biotin switch assay and western blot analysis, which revealed that UGDH and G6PDH were susceptible to SNO. Furthermore, mass spectrometry analysis of proteins treated with S-nitrosoglutathione (GSNO) identified the SNO modification sites for UGDH and G6PDH as cysteine 423 and cysteine 249, respectively. These findings suggest that NO modulates polysaccharide biosynthesis in N. flagelliforme through SNO of UGDH and G6PDH. This reveals a potential mechanism through which NO promotes polysaccharide synthesis in N. flagelliforme, while also providing a new strategy for improving the industrial production of polysaccharides.

基于水杨酸和茉莉酸通过调节细胞内一氧化氮(NO)水平增加鞭毛藻多糖产量的研究结果,我们从S-亚硝基化(SNO)的角度探讨了NO影响鞭毛藻多糖生物合成的机制。加入 NO 供体和清除剂后发现,细胞内 NO 对鞭毛藻多糖产量有显著的正向影响。为了探索其机制,我们研究了 NO 水平与参与多糖生物合成的几种关键酶活性之间的关系,包括 1,6-二磷酸果糖醛缩酶 (FBA)、葡萄糖激酶 (GK)、6-磷酸葡萄糖脱氢酶 (G6PDH)、线粒体异柠檬酸脱氢酶 (ICDH) 和 UDP-葡萄糖脱氢酶 (UGDH)。研究表明,G6PDH、ICDH 和 UGDH 的酶活性与细胞内 NO 水平的变化显著相关。为了进一步验证,在大肠杆菌中异源表达了 G6PDH、ICDH 和 UGDH,并通过 Ni+-NAT 亲和层析进行纯化,然后进行生物素转换测定和 Western 印迹分析,结果显示 UGDH 和 G6PDH 易受 SNO 影响。此外,经 S-亚硝基谷胱甘肽(GSNO)处理的蛋白质的质谱分析表明,UGDH 和 G6PDH 的 SNO 修饰位点分别为半胱氨酸 423 和半胱氨酸 249。这些发现表明,氮氧化物通过对 UGDH 和 G6PDH 的 SNO 来调节鞭毛虫多糖的生物合成。这揭示了 NO 促进鞭毛菜多糖合成的潜在机制,同时也为改善多糖的工业生产提供了新策略。
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引用次数: 0
Recombinant hirudin and PAR-1 regulate macrophage polarisation status in diffuse large B-cell lymphoma 重组水蛭素和 PAR-1 可调节弥漫大 B 细胞淋巴瘤中巨噬细胞的极化状态
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-12 DOI: 10.1186/s12896-024-00879-w
Qiang Pei, Zihui Li, Jingjing Zhao, Haixi Zhang, Tao Qin, Juan Zhao
Diffuse large B-cell lymphoma (DLBCL) is a malignant tumour. Although some standard therapies have been established to improve the cure rate, they remain ineffective for specific individuals. Therefore, it is meaningful to find more novel therapeutic approaches. Macrophage polarisation is extensively involved in the process of tumour development. Recombinant hirudin (rH) affects macrophages and has been researched frequently in clinical trials lately. Our article validated the regulatory role of rH in macrophage polarisation and the mechanism of PAR-1 by collecting clinical samples and subsequently establishing a cellular model to provide a scientifically supported perspective for discovering new therapeutic approaches. We assessed the expression of macrophage polarisation markers, cytokines and PAR-1 in clinical samples. We established a cell model by co-culture with THP-1 and OCI-Ly10 cell. We determined the degree of cell polarisation and expression of validation cytokines by flow cytometry, ELISA, and RT-qPCR to confirm the success of the cell model. Subsequently, different doses of rH were added to discover the function of rH on cell polarisation. We confirmed the mechanism of PAR-1 in macrophage polarisation by transfecting si-PAR-1 and pcDNA3.1-PAR-1. We found higher expression of M2 macrophage markers (CD163 + CMAF+) and PAR-1 in 32 DLBCL samples. After inducing monocyte differentiation into M0 macrophages and co-culturing with OCI-Ly10 lymphoma cells, we found a trend of these expressions in the cell model consistent with the clinical samples. Subsequently, we discovered that rH promotes the polarisation of M1 macrophages but inhibits the polarisation of M2 macrophages. We also found that PAR-1 regulates macrophage polarisation, inhibiting cell proliferation, migration, invasion and angiogenic capacity. rH inhibits macrophage polarisation towards the M2 type and PAR-1 regulates polarisation, proliferation, migration, invasion, and angiogenesis of DLBCL-associated macrophages.
弥漫大 B 细胞淋巴瘤(DLBCL)是一种恶性肿瘤。虽然一些标准疗法已被确立以提高治愈率,但它们对特定个体仍然无效。因此,寻找更多新的治疗方法意义重大。巨噬细胞极化广泛参与了肿瘤的发展过程。重组水蛭素(rH)会影响巨噬细胞,最近在临床试验中被频繁研究。我们的文章通过收集临床样本验证了 rH 在巨噬细胞极化中的调控作用以及 PAR-1 的作用机制,并随后建立了一个细胞模型,为发现新的治疗方法提供了一个有科学依据的视角。我们评估了临床样本中巨噬细胞极化标志物、细胞因子和 PAR-1 的表达。我们通过与 THP-1 和 OCI-Ly10 细胞共培养建立了细胞模型。我们通过流式细胞术、ELISA 和 RT-qPCR 测定了细胞极化的程度和验证细胞因子的表达,以确认细胞模型的成功。随后,我们加入了不同剂量的 rH,以发现 rH 对细胞极化的作用。我们通过转染 si-PAR-1 和 pcDNA3.1-PAR-1 证实了 PAR-1 在巨噬细胞极化中的作用机制。我们发现在32个DLBCL样本中,M2巨噬细胞标志物(CD163 + CMAF+)和PAR-1的表达较高。在诱导单核细胞分化为 M0 巨噬细胞并与 OCI-Ly10 淋巴瘤细胞共培养后,我们发现细胞模型中这些表达的趋势与临床样本一致。随后,我们发现 rH 可促进 M1 巨噬细胞的极化,但会抑制 M2 巨噬细胞的极化。我们还发现,PAR-1 可调节巨噬细胞的极化,抑制细胞的增殖、迁移、侵袭和血管生成能力。
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引用次数: 0
Comparison of lipidome profiles in serum from lactating dairy cows supplemented with Acremonium terrestris culture based on UPLC-QTRAP-MS/MS 基于 UPLC-QTRAP-MS/MS 的泌乳奶牛血清脂质体谱分析比较
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-12 DOI: 10.1186/s12896-024-00881-2
Chenmiao Zhang, Yiran Zhao, Shijiao Guo, Feifei Li, Xu Gong, Jiarui Gao, Linshu Jiang, Jinjin Tong
This study evaluated the effects of supplementing the diet of lactating cows with Acremonium terrestris culture (ATC) on milk production, serum antioxidant capacity, inflammatory indices, and serum lipid metabolomics. Over 90 days, 24 multiparous Chinese Holstein cows in mid-lactation (108 ± 10.4 days in milk, 637 ± 25 kg body weight, 30.23 ± 3.7 kg/d milk yield) were divided into either a control diet (CON) or a diet supplemented with 30 g of ATC daily. All the data were analyzed using Student’s t test with SPSS 20.0 software. The results showed that compared with CON feeding, ATC feeding significantly increased milk yield, antioxidant capacity, and immune function. Lipidome screening identified 143 lipid metabolites that differed between the two groups. Further analysis using “random forest” machine learning revealed three glycerophospholipid serum metabolites that could serve as lipid markers with a predictive accuracy of 91.67%. This study suggests that ATC can be a useful dietary supplement for improving lactational performance in dairy cows and provides valuable insights into developing nutritional strategies to maintain metabolic homeostasis in ruminants.
本研究评估了在泌乳奶牛的日粮中添加白藜芦醇培养物(ATC)对产奶量、血清抗氧化能力、炎症指数和血清脂质代谢组学的影响。在 90 天内,将 24 头处于泌乳中期的多胎中国荷斯坦奶牛(108 ± 10.4 天,637 ± 25 千克体重,30.23 ± 3.7 千克/天产奶量)分为对照日粮(CON)或每天添加 30 克 ATC 的日粮。所有数据均采用 SPSS 20.0 软件的学生 t 检验进行分析。结果表明,与对照组相比,ATC能显著提高产奶量、抗氧化能力和免疫功能。脂质体筛选确定了两组之间存在差异的 143 种脂质代谢物。利用 "随机森林 "机器学习进行的进一步分析发现,有三种甘油磷脂血清代谢物可作为脂质标记物,预测准确率高达 91.67%。这项研究表明,ATC 可以作为一种有用的膳食补充剂来提高奶牛的泌乳性能,并为制定维持反刍动物代谢平衡的营养策略提供了有价值的见解。
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引用次数: 0
Antimicrobial and cytotoxic activities of flavonoid and phenolics extracted from Sepia pharaonis ink (Mollusca: Cephalopoda) 从墨鱼(软体动物门:头足纲)中提取的类黄酮和酚类物质的抗菌和细胞毒性活性
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-12 DOI: 10.1186/s12896-024-00880-3
Asmaa R. Abdel-Malek, Alaa Y. Moustafa, Shimaa H. Salem
Several studies have been reported previously on the bioactivities of different extracts of marine molluscs. Therefore, we decided to evaluate the cytotoxic and antimicrobial activities of S. pharaonis ink as a highly populated species in the Red Sea. We extracted the flavonoids from the ink and analyzed their composition. Then we evaluated systematically the cytotoxic and antimicrobial properties of this extract. A pharmacokinetic study was also conducted using SwissADME to assess the potential of the identified flavonoids and phenolic compounds from the ink extract to be orally active drug candidates. Cytotoxic activity was evaluated against 5 cell lines (MCF7, Hep G2, A549, and Caco2) at different concentrations (0.4 µg/mL, 1.6 µg/mL, 6.3 µg/mL, 25 µg/mL, 100 µg/mL). The viability of examined cells was reduced by the extract in a concentration-dependent manner. The highest cytotoxic effect of the extract was recorded against A549 and Hep G2 cancer cell lines cells with IC50 = 2.873 and 7.1 µg/mL respectively. The mechanistic analysis by flow cytometry of this extract on cell cycle progression and apoptosis induction indicated that the extract arrests the cell cycle at the S phase in Hep G2 and MCF7, while in A549 cell arrest was recorded at G1 phase. However, it causes G1 and S phase arrest in Caco2 cancer cell line. Our data showed that the extract has significant antimicrobial activity against all tested human microbial pathogens. However, the best inhibitory effect was observed against Candida albicans ATCC 10,221 with a minimum inhibitory concentration (MIC) of 1.95 µg/mL. Pharmacokinetic analysis using SwissADME showed that most flavonoids and phenolics compounds have high drug similarity as they satisfy Lipinski’s criteria and have WLOGP values below 5.88 and TPSA below 131.6 Å2. S. pharaonis ink ethanolic extract showed a promising cytotoxic potency against various cell lines and a remarkable antimicrobial action against different pathogenic microbial strains. S. pharaonis ink is a novel source of important flavonoids that could be used in the future in different applications as a naturally safe and feasible alternative of synthetic drugs.
关于海洋软体动物不同萃取物的生物活性,此前已有多项研究报道。因此,我们决定评估红海中大量存在的 S. pharaonis 墨汁的细胞毒性和抗菌活性。我们从墨水中提取了黄酮类化合物,并分析了其成分。然后,我们系统地评估了这种提取物的细胞毒性和抗菌特性。我们还使用 SwissADME 进行了药代动力学研究,以评估从墨汁提取物中鉴定出的黄酮类化合物和酚类化合物作为口服活性药物候选物的潜力。在不同浓度(0.4 µg/mL、1.6 µg/mL、6.3 µg/mL、25 µg/mL、100 µg/mL)下,对 5 种细胞系(MCF7、Hep G2、A549 和 Caco2)的细胞毒活性进行了评估。萃取物以浓度依赖的方式降低了受检细胞的活力。萃取物对 A549 和 Hep G2 癌细胞系细胞的细胞毒性效果最高,IC50 分别为 2.873 和 7.1 µg/mL。流式细胞仪对该提取物的细胞周期进展和凋亡诱导机理分析表明,该提取物在 Hep G2 和 MCF7 中使细胞周期停滞在 S 期,而在 A549 中则使细胞停滞在 G1 期。然而,它在 Caco2 癌细胞系中会导致 G1 期和 S 期细胞周期的停滞。我们的数据显示,该提取物对所有测试的人类微生物病原体都有显著的抗菌活性。不过,对白色念珠菌(Candida albicans ATCC 10,221)的抑制效果最好,最低抑制浓度(MIC)为 1.95 µg/mL。使用 SwissADME 进行的药代动力学分析表明,大多数黄酮类和酚类化合物具有很高的药物相似性,因为它们符合利宾斯基标准,WLOGP 值低于 5.88,TPSA 值低于 131.6 Å2。S. pharaonis 墨汁乙醇提取物对多种细胞株具有良好的细胞毒性,对不同的病原微生物菌株具有显著的抗菌作用。法老墨汁是一种重要黄酮类化合物的新来源,未来可作为天然安全可行的合成药物替代品用于不同的应用领域。
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引用次数: 0
Biocompatibility and potential anticancer activity of gadolinium oxide (Gd2O3) nanoparticles against nasal squamous cell carcinoma. 氧化钆(Gd2O3)纳米颗粒对鼻腔鳞状细胞癌的生物相容性和潜在抗癌活性。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-06 DOI: 10.1186/s12896-024-00877-y
Xiaopeng Sun, Bo Kou

Chemotherapy as a cornerstone of cancer treatment is slowly being edged aside owing to its severe side effects and systemic toxicity. In this case, nanomedicine has emerged as an effective tool to address these drawbacks. Herein, a biocompatible carrier based on bovine serum albumin (BSA) coated gadolinium oxide nanoparticles (Gd2O3@BSA) was fabricated for curcumin (CUR) delivery and its physicochemical features along with its potential anticancer activity against nasal squamous cell carcinoma were also investigated. It was found that the fabricated Gd2O3@BSA containing CUR (Gd2O3@BSA-CUR) had spherical morphology with hydrodynamic size of nearly 26 nm, zeta-potential of -36 mV and high drug (CUR) loading capacity. Drug release profile disclosed that the release of CUR from the prepared Gd2O3@BSA-CUR nanoparticles occurred in a sustained- and pH-dependent manner. Also, in vitro cytotoxicity analysis revealed that the fabricated Gd2O3@BSA nanoparticles possessed excellent biosafety toward HFF2 normal cells, while Gd2O3@BSA-CUR appeared to display the greatest anticancer potential against RPMI 2650 and CNE-1 cancer cell lines. The results also show that the Gd2O3@BSA nanoparticles were compatible with the blood cells with minor hemolytic effect (< 3%). The manufactured NPs were found to be completely safe for biological applications in an in vivo subacute toxicity study. Taken together, these finding substantiate the potential anticancer activity of Gd2O3@BSA-CUR nanoparticles against nasal squamous cell carcinoma, but the results obtained demand further studies to assess their full potential.

化疗作为癌症治疗的基石,由于其严重的副作用和全身毒性,正逐渐被边缘化。在这种情况下,纳米药物已成为解决这些弊端的有效工具。本文制备了一种基于牛血清白蛋白(BSA)包覆氧化钆纳米颗粒(Gd2O3@BSA)的生物相容性载体,用于姜黄素(CUR)的递送,并研究了其理化特性及其对鼻腔鳞状细胞癌的潜在抗癌活性。研究发现,所制备的含有姜黄素的 Gd2O3@BSA (Gd2O3@BSA-CUR)具有球形形态,流体力学尺寸接近 26 nm,zeta 电位为 -36 mV,具有较高的药物(姜黄素)负载能力。药物释放曲线显示,CUR 从制备的 Gd2O3@BSA-CUR 纳米粒子中的释放是以持续和 pH 值依赖的方式进行的。体外细胞毒性分析表明,制备的 Gd2O3@BSA 纳米粒子对 HFF2 正常细胞具有良好的生物安全性,而 Gd2O3@BSA-CUR 对 RPMI 2650 和 CNE-1 癌细胞株的抗癌潜力最大。研究结果还表明,Gd2O3@BSA 纳米粒子与血细胞相容,具有轻微的溶血效应(2O3@BSA-CUR 纳米粒子对鼻腔鳞状细胞癌具有抗癌作用,但还需要进一步研究才能评估其全部潜力)。
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引用次数: 0
In vitro evaluation of PLGA loaded hesperidin on colorectal cancer cell lines: an insight into nano delivery system. 聚乳酸乙烯雌酚负载橙皮甙对结直肠癌细胞系的体外评估:对纳米给药系统的深入了解。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-02 DOI: 10.1186/s12896-024-00882-1
Narges Yaghoubi, Amir Gholamzad, Tahere Naji, Mehrdad Gholamzad

Background: Colorectal cancer is a common disease worldwide with non-specific symptoms such as blood in the stool, bowel movements, weight loss and fatigue. Chemotherapy drugs can cause side effects such as nausea, vomiting and a weakened immune system. The use of antioxidants such as hesperidin could reduce the side effects, but its low bioavailability is a major problem. In this research, we aimed to explore the drug delivery and efficiency of this antioxidant on the HCT116 colorectal cancer cell line by loading hesperidin into PLGA nanoparticles.

Materials and methods: Hesperidin loaded PLGA nanoparticles were produced by single emulsion evaporation method. The physicochemical properties of the synthesized hesperidin-loaded nanoparticles were determined using SEM, AFM, FT-IR, DLS and UV-Vis. Subsequently, the effect of the PLGA loaded hesperidin nanoparticles on the HCT116 cell line after 48 h was investigated by MTT assay at three different concentrations of the nanoparticles.

Result: The study showed that 90% of hesperidin were loaded in PLGA nanoparticles by UV-Vis spectrophotometry and FT-IR spectrum. The nanoparticles were found to be spherical and uniform with a hydrodynamic diameter of 76.2 nm in water. The release rate of the drug was about 93% after 144 h. The lowest percentage of cell viability of cancer cells was observed at a concentration of 10 µg/ml of PLGA nanoparticles loaded with hesperidin.

Conclusion: The results indicate that PLGA nanoparticles loaded with hesperidin effectively reduce the survival rate of HCT116 colorectal cancer cells. However, further studies are needed to determine the appropriate therapeutic dosage and to conduct animal and clinical studies.

背景:结肠直肠癌是一种全球常见疾病,患者会出现便血、大便次数增多、体重减轻和疲劳等非特异性症状。化疗药物会导致恶心、呕吐和免疫力下降等副作用。使用橙皮甙等抗氧化剂可减轻副作用,但其生物利用率低是一个主要问题。本研究旨在通过将橙皮甙载入聚乳酸(PLGA)纳米颗粒,探讨这种抗氧化剂对 HCT116 大肠癌细胞株的药物输送和效率。用扫描电镜、原子力显微镜、傅立叶变换红外光谱、激光光度计和紫外可见光谱测定了合成的橙皮甙负载纳米粒子的理化性质。随后,采用 MTT 法研究了三种不同浓度的负载橙皮甙的 PLGA 纳米颗粒在 48 小时后对 HCT116 细胞系的影响:研究结果:紫外-可见分光光度法和傅立叶变换红外光谱显示,90%的橙皮甙被载入 PLGA 纳米颗粒。纳米颗粒呈均匀的球形,在水中的流体力学直径为 76.2 nm。144小时后,药物释放率约为93%。在浓度为10 µg/ml的PLGA纳米颗粒中添加橙皮甙后,癌细胞存活率最低:结果表明,负载橙皮甙的 PLGA 纳米颗粒能有效降低 HCT116 大肠癌细胞的存活率。然而,还需要进一步研究确定适当的治疗剂量,并开展动物和临床研究。
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引用次数: 0
Ferula latisecta gels for synthesis of zinc/silver binary nanoparticles: antibacterial effects against gram-negative and gram-positive bacteria and physicochemical characteristics 用于合成锌/银二元纳米粒子的阿魏凝胶:对革兰氏阴性菌和革兰氏阳性菌的抗菌效果及理化特性
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-08-01 DOI: 10.1186/s12896-024-00878-x
Ali Es-haghi, Mohammad Sadegh Amiri, Mohammad Ehsan Taghavizadeh Yazdi
This study explores the potential antibacterial applications of zinc oxide nanoparticles (ZnO NPs) enhanced with silver (Ag) using plant gel (ZnO-AgO NPs). The problem addressed is the increasing prevalence of pathogenic bacteria and the need for new, effective antimicrobial agents. ZnO NPs possess distinctive physicochemical properties that enable them to selectively target bacterial cells. Their small size and high surface area-to-volume ratio allow efficient cellular uptake and interaction with bacterial cells. In this study, the average size of the synthesized ZnO-Ag nanoparticles was 77.1 nm, with a significant standard deviation of 33.7 nm, indicating a wide size distribution. The nanoparticles demonstrated remarkable antibacterial efficacy against gram-negative and gram-positive bacteria, with inhibition zones of 14.33 mm for E. coli and 15.66 mm for B. subtilis at a concentration of 300 µg/ml. Minimum inhibitory concentrations (MIC) were determined to be 100 µg/ml for E. coli and 75 µg/ml for S. saprophyticus. Additionally, ZnO-Ag NPs exhibited excellent biocompatibility, making them appropriate for various pharmacological uses. This study utilizes Ferula latisecta gels, offering a sustainable and eco-friendly approach to nanoparticle synthesis. Incorporating of Ag into ZnO NPs significantly enhances their antimicrobial properties, with the combined results showing great inhibition effects on pathogenic microbes. The findings suggest that ZnO-Ag NPs could be a promising candidate for addressing the challenges posed by drug-resistant bacterial infections and enhancing antimicrobial treatments.
本研究探讨了利用植物凝胶(ZnO-AgO NPs)增强银(Ag)的氧化锌纳米粒子(ZnO NPs)的潜在抗菌应用。所要解决的问题是病原菌的日益流行以及对新型有效抗菌剂的需求。氧化锌氮氧化物具有独特的物理化学特性,能够选择性地靶向细菌细胞。它们的尺寸小、表面积与体积比高,能被细胞有效吸收并与细菌细胞相互作用。在这项研究中,合成的 ZnO-Ag 纳米粒子的平均尺寸为 77.1 nm,标准偏差为 33.7 nm,表明其尺寸分布较广。纳米粒子对革兰氏阴性菌和革兰氏阳性菌具有显著的抗菌效果,在浓度为 300 µg/ml 时,对大肠杆菌和枯草杆菌的抑菌区分别为 14.33 mm 和 15.66 mm。对大肠杆菌的最小抑菌浓度(MIC)为 100 微克/毫升,对沙门氏菌的最小抑菌浓度(MIC)为 75 微克/毫升。此外,ZnO-Ag NPs 还具有良好的生物相容性,因此适合用于各种药理用途。这项研究利用了Ferula latisecta凝胶,为纳米粒子合成提供了一种可持续的环保方法。在 ZnO NPs 中加入 Ag 可显著增强其抗菌性能,综合结果显示对病原微生物有很好的抑制作用。研究结果表明,ZnO-Ag NPs 有望成为应对耐药细菌感染挑战和加强抗菌治疗的候选材料。
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引用次数: 0
Validation of a rapid collagenase activity detection technique based on fluorescent quenched gelatin with synovial fluid samples. 利用滑膜液样本验证基于荧光淬灭明胶的胶原酶活性快速检测技术。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-07-19 DOI: 10.1186/s12896-024-00869-y
Miguel de la Fuente, Diego Delgado, Maider Beitia, Gabriel Barreda-Gómez, Arantxa Acera, Mikel Sanchez, Elena Vecino

Background: Measuring collagenase activity is crucial in the field of joint health and disease management. Collagenases, enzymes responsible for collagen degradation, play a vital role in maintaining the balance between collagen synthesis and breakdown in joints. Dysregulation of collagenase activity leads to joint tissue degradation and diseases such as rheumatoid arthritis and osteoarthritis. The development of methods to measure collagenase activity is essential for diagnosis, disease severity assessment, treatment monitoring, and identification of therapeutic targets.

Results: This study aimed to validate a rapid collagenase activity detection technique using synovial fluid samples. Antibody microarray analysis was initially performed to quantify the levels of matrix metalloproteinase-9 (MMP-9), a major collagenase in joints. Subsequently, the developed gelatin-based test utilizing fluorescence measurement was used to determine collagenase activity. There was a significant correlation between the presence of MMP-9 and collagenase activity. In addition, Lower Limit of Detection and Upper Limit of Detection can be preliminary estimated as 8 ng/mL and 48 ng/mL respectively.

Conclusions: The developed technique offers a potential point-of-care assessment of collagenase activity, providing real-time information for clinicians and researchers. By accurately quantifying collagenase activity, healthcare professionals can optimize patient care, improve treatment outcomes, and contribute to the understanding and management of joint-related disorders. Further research and validation are necessary to establish the full potential of this rapid collagenase activity detection method in clinical practice.

背景:在关节健康和疾病管理领域,测量胶原酶活性至关重要。胶原蛋白酶是一种负责降解胶原蛋白的酶,在维持关节中胶原蛋白合成和分解之间的平衡方面起着至关重要的作用。胶原酶活性失调会导致关节组织退化,引发类风湿性关节炎和骨关节炎等疾病。开发测量胶原酶活性的方法对于诊断、疾病严重程度评估、治疗监测和确定治疗目标至关重要:本研究旨在利用滑液样本验证胶原酶活性快速检测技术。首先进行了抗体微阵列分析,以量化关节中主要胶原酶基质金属蛋白酶-9(MMP-9)的水平。随后,利用荧光测量法开发的明胶检测法确定胶原酶的活性。MMP-9 的存在与胶原酶活性之间存在明显的相关性。此外,初步估计检测下限和检测上限分别为 8 纳克/毫升和 48 纳克/毫升:所开发的技术提供了一种潜在的胶原酶活性护理点评估方法,为临床医生和研究人员提供了实时信息。通过准确量化胶原蛋白酶的活性,医护人员可以优化患者护理、改善治疗效果,并促进对关节相关疾病的理解和管理。要充分发挥这种胶原蛋白酶活性快速检测方法在临床实践中的潜力,还需要进一步的研究和验证。
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引用次数: 0
Characterization of a thermostable protease from Bacillus subtilis BSP strain. 枯草芽孢杆菌 BSP 菌株的一种恒温蛋白酶的特性。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-07-15 DOI: 10.1186/s12896-024-00870-5
Tanveer Majeed, Charles C Lee, William J Orts, Romana Tabassum, Tawaf Ali Shah, Yousef A Bin Jardan, Turki M Dawoud, Mohammed Bourhia

This study used conservative one variable-at-a-time study and statistical surface response methods to increase the yields of an extracellular thermostable protease secreted by a newly identified thermophilic Bacillus subtilis BSP strain. Using conventional optimization techniques, physical parameters in submerged fermentation were adjusted at the shake flask level to reach 184 U/mL. These physicochemical parameters were further optimized by statistical surface response methodology using Box Behnken design, and the protease yield increased to 295 U/mL. The protease was purified and characterized biochemically. Both Ca2+ and Fe2+ increased the activity of the 36 kDa protease enzyme. Based on its strong inhibition by ethylenediaminetetracetate (EDTA), the enzyme was confirmed to be a metalloprotease. The protease was also resistant to various organic solvents (benzene, ethanol, methanol), surfactants (Triton X-100), sodium dodecyl sulfate (SDS), Tween 20, Tween-80 and oxidants hydrogen per oxide (H2O2). Characteristics, such as tolerance to high SDS and H2O2 concentrations, indicate that this protease has potential applications in the pharmaceutical and detergent industries.

本研究采用保守的一次一变量研究和统计表面响应方法来提高新发现的嗜热枯草芽孢杆菌 BSP 菌株分泌的胞外恒温蛋白酶的产量。利用传统的优化技术,在摇瓶水平上调整了浸没发酵的物理参数,使其达到 184 U/mL。利用盒式贝肯设计(Box Behnken design)的统计表面响应方法进一步优化了这些理化参数,蛋白酶产量提高到 295 U/mL。对蛋白酶进行了纯化和生化鉴定。Ca2+ 和 Fe2+ 都能提高 36 kDa 蛋白酶的活性。根据乙二胺四乙酸(EDTA)对该酶的强烈抑制作用,证实该酶是一种金属蛋白酶。该蛋白酶还能抵抗各种有机溶剂(苯、乙醇、甲醇)、表面活性剂(Triton X-100)、十二烷基硫酸钠(SDS)、吐温 20、吐温-80 和氧化剂一氧化氢(H2O2)。它对高浓度 SDS 和 H2O2 的耐受性等特性表明,这种蛋白酶有可能应用于制药和洗涤剂行业。
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引用次数: 0
Recombinant human enamelin produced in Escherichia coli promotes mineralization in vitro. 在大肠杆菌中生产的重组人牙釉质素能促进体外矿化。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-07-09 DOI: 10.1186/s12896-024-00875-0
Monalissa Halablab, Lovisa Wallman, Johan Bonde

Background: Enamelin is an enamel matrix protein that plays an essential role in the formation of enamel, the most mineralized tissue in the human body. Previous studies using animal models and proteins from natural sources point to a key role of enamelin in promoting mineralization events during enamel formation. However, natural sources of enamelin are scarce and with the current study we therefore aimed to establish a simple microbial production method for recombinant human enamelin to support its use as a mineralization agent.

Results: In the study the 32 kDa fragment of human enamelin was successfully expressed in Escherichia coli and could be obtained using immobilized metal ion affinity chromatography purification (IMAC), dialysis, and lyophilization. This workflow resulted in a yield of approximately 10 mg enamelin per liter culture. Optimal conditions for IMAC purification were obtained using Ni2+ as the metal ion, and when including 30 mM imidazole during binding and washing steps. Furthermore, in vitro mineralization assays demonstrated that the recombinant enamelin could promote calcium phosphate mineralization at a concentration of 0.5 mg/ml.

Conclusions: These findings address the scarcity of enamelin by facilitating its accessibility for further investigations into the mechanism of enamel formation and open new avenues for developing enamel-inspired mineralized biomaterials.

背景:牙釉质素是一种牙釉质基质蛋白,在人体矿化度最高的组织--牙釉质的形成过程中发挥着至关重要的作用。以往利用动物模型和天然来源蛋白质进行的研究表明,在釉质形成过程中,牙釉质素在促进矿化过程中起着关键作用。然而,天然来源的牙釉质素非常稀缺,因此我们目前的研究旨在建立一种简单的微生物生产重组人牙釉质素的方法,以支持其作为矿化剂的使用:在这项研究中,32 kDa 的人牙釉质素片段成功地在大肠杆菌中表达,并可通过固定金属离子亲和层析纯化(IMAC)、透析和冻干获得。通过这一工作流程,每升培养物可获得约 10 毫克牙釉质素。使用 Ni2+ 作为金属离子,并在结合和洗涤步骤中加入 30 mM 的咪唑时,获得了 IMAC 纯化的最佳条件。此外,体外矿化试验表明,浓度为 0.5 mg/ml 的重组牙釉质素能促进磷酸钙矿化:这些发现解决了牙釉质素稀缺的问题,使人们更容易获得牙釉质素,从而进一步研究牙釉质的形成机制,并为开发受牙釉质启发的矿化生物材料开辟了新途径。
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