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Preparation and preliminary application of fluorescent microsphere test strips for feline parvovirus antibodies. 猫细小病毒抗体荧光微球试纸的制备和初步应用。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2024-09-27 DOI: 10.1186/s12896-024-00900-2
Jinyuan Shang, Manping Yan, Xiaohao Zhang, Wei Liu, Shun Wu, Zhenjun Wang, Li Yi, Chunxia Wang, Erkai Feng, Yuening Cheng, Guoliang Luo
{"title":"Preparation and preliminary application of fluorescent microsphere test strips for feline parvovirus antibodies.","authors":"Jinyuan Shang, Manping Yan, Xiaohao Zhang, Wei Liu, Shun Wu, Zhenjun Wang, Li Yi, Chunxia Wang, Erkai Feng, Yuening Cheng, Guoliang Luo","doi":"10.1186/s12896-024-00900-2","DOIUrl":"https://doi.org/10.1186/s12896-024-00900-2","url":null,"abstract":"<p><p>This study introduces a novel diagnostic modality for the detection of feline panleukopenia virus (FPV) antibodies in feline serum by using fluorescent microsphere immunochromatographic test strips (FM-ICTS). Leveraging the inherent specificity of antigen-antibody interactions, the FM-ICTS approach demonstrates considerable potential for efficient and accurate FPV antibody detection within a short timeframe. The FM-ICTS method demonstrates strong diagnostic performance, with consistent accuracy and stability over time. PBS buffer dilution enables detection across the range of FPV antibody haemagglutination inhibition (HI) titres in both healthy and immunized or infected cats. A high correlation (R² = 0.9733) between the T/C ratio and FPV antibody titres confirms the method's effectiveness in quantifying these titres. Clinical validation with 84 samples supports its reliability by matching results with HI assays. Additionally, stability tests show that the test strips maintain performance during storage, with a coefficient of variation (CV) below 12% over three months at 25℃. This innovative FM-ICTS framework emerges as a promising avenue for expedient and dependable disease diagnosis within the realm of veterinary science, offering implications for timely disease management and surveillance.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"65"},"PeriodicalIF":3.5,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11429854/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142340570","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Systematic design and evaluation of aptamers for VEGF and PlGF biomarkers of Preeclampsia. 系统设计和评估子痫前期血管内皮生长因子和 PlGF 生物标记物的适配体。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2024-09-27 DOI: 10.1186/s12896-024-00891-0
Samavath Mallawarachchi, Rümeysa E Cebecioglu, Majed Althumayri, Levent Beker, Sandun Fernando, Hatice Ceylan Koydemir
{"title":"Systematic design and evaluation of aptamers for VEGF and PlGF biomarkers of Preeclampsia.","authors":"Samavath Mallawarachchi, Rümeysa E Cebecioglu, Majed Althumayri, Levent Beker, Sandun Fernando, Hatice Ceylan Koydemir","doi":"10.1186/s12896-024-00891-0","DOIUrl":"https://doi.org/10.1186/s12896-024-00891-0","url":null,"abstract":"<p><p>Preeclampsia is a potentially life-threatening condition for both mother and baby, characterized by hypertension and potential organ damage. Early diagnosis is crucial to mitigate its adverse health effects. Traditional diagnostic methods, which focus on late-manifesting symptoms like hypertension and proteinuria, underscore the need for molecular diagnostic approaches for timely detection. This study successfully designs and evaluates novel aptamers with high specificity and affinity for Vascular Endothelial Growth Factor (VEGF) and Placental Growth Factor (PlGF), biomarkers closely associated with preeclampsia. Using molecular docking, molecular dynamics simulations, and BioLayer Interferometry (BLI), we identified aptamers that demonstrated strong binding affinities, comparable or superior to traditional antibodies. Our findings suggest that these aptamers have the potential to be integrated into cost-effective, point-of-care diagnostic tools, significantly improving early detection and intervention strategies for preeclampsia. The robust performance of these aptamers marks a pivotal step toward the development of more reliable and accessible diagnostic solutions, with implications for better maternal and fetal health outcomes.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"64"},"PeriodicalIF":3.5,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11428563/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142340571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Unraveling the impact of pH, sodium concentration, and medium osmolality on Vibrio natriegens in batch processes. 揭示批处理过程中 pH 值、钠浓度和培养基渗透压对纳氏弧菌的影响。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2024-09-23 DOI: 10.1186/s12896-024-00897-8
Eva Forsten, Steffen Gerdes, René Petri, Jochen Büchs, Jørgen Magnus
{"title":"Unraveling the impact of pH, sodium concentration, and medium osmolality on Vibrio natriegens in batch processes.","authors":"Eva Forsten, Steffen Gerdes, René Petri, Jochen Büchs, Jørgen Magnus","doi":"10.1186/s12896-024-00897-8","DOIUrl":"10.1186/s12896-024-00897-8","url":null,"abstract":"<p><strong>Background: </strong>Vibrio natriegens, a halophilic marine γ-proteobacterium, holds immense biotechnological potential due to its remarkably short generation time of under ten minutes. However, the highest growth rates have been primarily observed on complex media, which often suffer from batch-to-batch variability affecting process stability and performance. Consistent bioprocesses necessitate the use of chemically defined media, which are usually optimized for fermenters with pH and dissolved oxygen tension (DOT) regulation, both of which are not applied during early-stage cultivations in shake flasks or microtiter plates. Existing studies on V. natriegens' growth on mineral media report partially conflicting results, and a comprehensive study examining the combined effects of pH buffering, sodium concentration, and medium osmolality is lacking.</p><p><strong>Results: </strong>This study evaluates the influence of sodium concentration, pH buffering, and medium osmolality on the growth of V. natriegens under unregulated small-scale conditions. The maximum growth rate, time of glucose depletion, as well as the onset of stationary phase were observed through online-monitoring the oxygen transfer rate. The results revealed optimal growth conditions at an initial pH of 8.0 with a minimum of 300 mM MOPS buffer for media containing 20 g/L glucose or 180 mM MOPS for media with 10 g/L glucose. Optimal sodium chloride supplementation was found to be between 7.5 and 15 g/L, lower than previously reported ranges. This is advantageous for reducing industrial corrosion issues. Additionally, an osmolality range of 1 to 1.6 Osmol/kg was determined to be optimal for growth. Under these optimized conditions, V. natriegens achieved a growth rate of 1.97 ± 0.13 1/h over a period of 1 h at 37 °C, the highest reported rate for this organism on a mineral medium.</p><p><strong>Conclusion: </strong>This study provides guidelines for cultivating V. natriegens in early-stage laboratory settings without pH and DOT regulation. The findings suggest a lower optimal sodium chloride range than previously reported and establish an osmolality window for optimal growth, thereby advancing the understanding of V. natriegens' physiology. In addition, this study offers a foundation for future research into the effects of different ions and carbon sources on V. natriegens.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"63"},"PeriodicalIF":3.5,"publicationDate":"2024-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11421182/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142307070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Anti-inflammatory potential of aspergillus unguis SP51-EGY: TLR4-dependent effects & chemical diversity via Q-TOF LC-HRMS unguis SP51-EGY 的抗炎潜力:通过 Q-TOF LC-HRMS 分析 TLR4 依赖性效应和化学多样性
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2024-09-18 DOI: 10.1186/s12896-024-00890-1
Soad Nasr, Abdelhameed S. Dawood, Amal Mosad Ibrahim, Mohamed S. Abdel-Aziz, Walid Fayad, Anwar Abdelnaser, Faten K. Abd EL-Hady
{"title":"Anti-inflammatory potential of aspergillus unguis SP51-EGY: TLR4-dependent effects & chemical diversity via Q-TOF LC-HRMS","authors":"Soad Nasr, Abdelhameed S. Dawood, Amal Mosad Ibrahim, Mohamed S. Abdel-Aziz, Walid Fayad, Anwar Abdelnaser, Faten K. Abd EL-Hady","doi":"10.1186/s12896-024-00890-1","DOIUrl":"https://doi.org/10.1186/s12896-024-00890-1","url":null,"abstract":"Inflammation serves as an intricate defense mechanism for tissue repair. However, overactivation of TLR4-mediated inflammation by lipopolysaccharide (LPS) can lead to detrimental outcomes such as sepsis, acute lung injury, and chronic inflammation, often associated with cancer and autoimmune diseases. This study delves into the anti-inflammatory properties of “Aspergillus unguis isolate SP51-EGY” on LPS-stimulated RAW 264.7 macrophages. Through real-time qPCR, we assessed the expression levels of pivotal inflammatory genes, including iNOS, COX-2, TNF-α, and IL-6. Remarkably, our fungal extracts significantly diminished NO production and showed noteworthy reductions in the mRNA expression levels of the aforementioned genes. Furthermore, while Nrf2 is typically associated with modulating inflammatory responses, our findings indicate that the anti-inflammatory effects of our extracts are not Nrf2-dependent. Moreover, the chemical diversity of the potent extract (B Sh F) was elucidated using Q-TOF LC-HRMS, identifying 54 compounds, some of which played vital roles in suppressing inflammation. Most notably, compounds like granisetron, fenofibrate, and umbelliprenin were found to downregulate TNF-α, IL-1β, and IL-6 through the NF-κB signaling pathway. In conclusion, “Aspergillus unguis isolate SP51-EGY”, isolated from the Red Sea, Egypt, has been unveiled as a promising TLR4 inhibitor with significant anti-inflammatory potentials, presenting novel insights for their potential therapeutic use in inflammation. ","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"35 1","pages":""},"PeriodicalIF":3.5,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142261040","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Synthesis and evaluation of nanosized aluminum MOF encapsulating Umbelliferon: assessing antioxidant, anti-inflammatory, and wound healing potential in an earthworm model 包覆伞形酮的纳米铝 MOF 的合成与评估:在蚯蚓模型中评估抗氧化、抗炎和伤口愈合潜力
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2024-09-15 DOI: 10.1186/s12896-024-00889-8
Rabab M. Thabit, Fatma El-Zahraa A. Abd El-Aziz, A. Abu El-Fadl, A. A. Abu-Sehly, Ahmed M. Sayed
{"title":"Synthesis and evaluation of nanosized aluminum MOF encapsulating Umbelliferon: assessing antioxidant, anti-inflammatory, and wound healing potential in an earthworm model","authors":"Rabab M. Thabit, Fatma El-Zahraa A. Abd El-Aziz, A. Abu El-Fadl, A. A. Abu-Sehly, Ahmed M. Sayed","doi":"10.1186/s12896-024-00889-8","DOIUrl":"https://doi.org/10.1186/s12896-024-00889-8","url":null,"abstract":"Nanoporous aluminum metal–organic framework (Al-MOF) was synthesized via solvothermal methods and employed as a carrier matrix for in vitro drug delivery of Umbelliferon (Um). The encapsulated Um was gradually released over seven days at 37 °C, using simulated body fluid phosphate-buffered saline (PBS) at pH 7.4 as the release medium. The drug release profile suggests the potential of Al-MOF nanoparticles as effective drug delivery carriers. Structural and chemical analyses of Um-loaded Al-MOF nanoparticles (Um-Al MOF) were conducted using Fourier-transform infrared (FTIR) spectroscopy, X-ray diffractometry (XRD), and ultraviolet–visible (UV–Vis) spectroscopy. Thermal gravimetric analysis (TGA) was employed to investigate the thermal stability of the Al-MOF nanoparticles, while Transmission Electron Microscopy (TEM) was utilized to assess their morphological features. Um-Al MOF nanoparticles demonstrated notable antioxidant and anti-inflammatory properties compared to Um and Al-MOF nanoparticles individually. Moreover, they exhibited significant enhancement in wound healing in an earthworm model. These findings underscore the potential of Al-MOF nanoparticles as a promising drug delivery system, necessitating further investigations to explore their clinical applicability.\u0000","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"90 1","pages":""},"PeriodicalIF":3.5,"publicationDate":"2024-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142261036","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A marker-free genetic manipulation method for Glaesserella parasuis strains developed by alternately culturing transformants at 37°C and 30°C. 通过在 37°C 和 30°C 温度下交替培养转化株,开发出一种无标记的寄生褐藻菌株遗传操作方法。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2024-09-03 DOI: 10.1186/s12896-024-00887-w
Jing Xiao, Yuxin Wang, Dongfang Wu, Yuping Song, Xuwang Cai, Huanchun Chen, Hongbo Zhou, Xiaojuan Xu
{"title":"A marker-free genetic manipulation method for Glaesserella parasuis strains developed by alternately culturing transformants at 37°C and 30°C.","authors":"Jing Xiao, Yuxin Wang, Dongfang Wu, Yuping Song, Xuwang Cai, Huanchun Chen, Hongbo Zhou, Xiaojuan Xu","doi":"10.1186/s12896-024-00887-w","DOIUrl":"10.1186/s12896-024-00887-w","url":null,"abstract":"<p><strong>Background: </strong>Glaesserella parasuis (G. parasuis) is the causative agent of Glässer's disease, which causes significant economic losses in the swine industry. However, research on the pathogenesis of G. parasuis has been hampered by the lack of a simple and efficient marker-free knockout system.</p><p><strong>Results: </strong>In this study, a marker-free knockout system was developed for G. parasuis using a temperature-sensitive vector. By alternating the incubation of transformants at 30°C and 37°C, we optimized the screening process for this system. The system was successfully applied to knockout the Kan<sup>R</sup> cassette from JS0135ΔnanH::Kan<sup>R</sup>, achieving a knockout efficiency of 90% in the final round of screening. To confirm that temperature variation was a key factor, we proceeded with knocking out the nanH and apd genes in the CF7066 strain. The knockout efficiency reached up to 100%, with the shortest screening time being only four days. The knockout of the nanH gene resulted in a significant reduction in the growth vitality of the strains, while the knockout of the apd gene led to an approximate 56% improvement in the adhesion rate. Additionally, we observed that the expression of recombinant genes in transformants was higher at 30℃ than at 37℃, with the recC gene being upregulated approximately 7-fold. In contrast, there was almost no difference in the expression of recombinant genes between 30℃ and 37℃ in the wild-type strains. This discrepancy was likely due to an elevated copy number of target plasmids at 30℃, which may have resulted in the enhanced expression of recombinant genes.</p><p><strong>Conclusions: </strong>In conclusion, this newly developed gene knockout system for G. parasuis presents a valuable tool for advancing research on this organism.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"60"},"PeriodicalIF":3.5,"publicationDate":"2024-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11373133/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142124721","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Purification and characterization of soluble recombinant Crimean-Congo hemorrhagic fever virus glycoprotein Gc expressed in mammalian 293F cells. 在哺乳动物 293F 细胞中表达的可溶性重组克里米亚-刚果出血热病毒糖蛋白 Gc 的纯化和表征。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2024-08-27 DOI: 10.1186/s12896-024-00885-y
Nigel Aminake Makoah, Matefo Millicent Litabe, Fredy Brice Nemg Simo, Katlego Keith Maboho, Felicity Jane Burt
{"title":"Purification and characterization of soluble recombinant Crimean-Congo hemorrhagic fever virus glycoprotein Gc expressed in mammalian 293F cells.","authors":"Nigel Aminake Makoah, Matefo Millicent Litabe, Fredy Brice Nemg Simo, Katlego Keith Maboho, Felicity Jane Burt","doi":"10.1186/s12896-024-00885-y","DOIUrl":"10.1186/s12896-024-00885-y","url":null,"abstract":"<p><strong>Background: </strong>Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne zoonotic disease that presents with severe hemorrhagic manifestations and is associated with significant fatality rates. The causative agent, Crimean-Congo Hemorrhagic Fever Virus (CCHFV), is a high-priority pathogen identified by the World Health Organization with no approved vaccine or specific treatment available. In addition, there is a critical need for enhanced diagnostic tools to improve public health awareness, prevention measures, and disease control strategies.</p><p><strong>Methods: </strong>We designed plasmids to enable the purification of soluble CCHFV glycoprotein Gc expressed in mammalian 293 F cells, followed by purification using affinity and size exclusion chromatography. The purified antigen was analyzed by SDS-PAGE and Western blotting to confirm its reactivity to antibodies from CCHF survivors. Additionally, an in-house indirect ELISA was developed using the purified Gc as a coating antigen.</p><p><strong>Results: </strong>The optimized expression system successfully produced soluble and pure Gc antigen after affinity chromatography. The protein showed specific reactivity with CCHFV-positive serum antibodies in Western blot analysis. The indirect ELISA assay demonstrated high efficacy in distinguishing between CCHFV-positive and -negative serum samples, indicating its potential as a valuable diagnostic tool. Size exclusion chromatography further confirmed the presence of aggregates in our protein preparation.</p><p><strong>Conclusions: </strong>The purified Gc antigen shows promise for developing direct diagnostic assays for CCHFV. The antigen's suitability for subunit vaccine development and its application as bait for monoclonal antibody isolation from survivors could be investigated further. This work lays the foundation for future research into the development of rapid diagnostic tests for field deployment.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"59"},"PeriodicalIF":3.5,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11348531/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142078967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Correction: Comparison of lipidome profiles in serum from lactating dairy cows supplemented with Acremonium terrestris culture based on UPLC-QTRAP-MS/MS. 更正:基于 UPLC-QTRAP-MS/MS 的泌乳奶牛血清脂质体图谱比较。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2024-08-22 DOI: 10.1186/s12896-024-00886-x
Chenmiao Zhang, Yiran Zhao, Shijiao Guo, Feifei Li, Xu Gong, Jiarui Gao, Linshu Jiang, Jinjin Tong
{"title":"Correction: Comparison of lipidome profiles in serum from lactating dairy cows supplemented with Acremonium terrestris culture based on UPLC-QTRAP-MS/MS.","authors":"Chenmiao Zhang, Yiran Zhao, Shijiao Guo, Feifei Li, Xu Gong, Jiarui Gao, Linshu Jiang, Jinjin Tong","doi":"10.1186/s12896-024-00886-x","DOIUrl":"10.1186/s12896-024-00886-x","url":null,"abstract":"","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"57"},"PeriodicalIF":3.5,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11361175/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142091842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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
BMC Biotechnology 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
{"title":"Nitric oxide mediates positive regulation of Nostoc flagelliforme polysaccharide yield via potential S-nitrosylation of G6PDH and UGDH.","authors":"Meng-Yuan Li, Yan-Ru Li, Cheng-Feng Han, Jie Zhang, Rui-Ying Zhu, Yan Zhang, Jian Li, Shi-Ru Jia, Pei-Pei Han","doi":"10.1186/s12896-024-00884-z","DOIUrl":"10.1186/s12896-024-00884-z","url":null,"abstract":"<p><p>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<sup>+</sup>-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.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"24 1","pages":"58"},"PeriodicalIF":3.5,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11342573/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142035140","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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
BMC Biotechnology Pub Date : 2024-08-12 DOI: 10.1186/s12896-024-00879-w
Qiang Pei, Zihui Li, Jingjing Zhao, Haixi Zhang, Tao Qin, Juan Zhao
{"title":"Recombinant hirudin and PAR-1 regulate macrophage polarisation status in diffuse large B-cell lymphoma","authors":"Qiang Pei, Zihui Li, Jingjing Zhao, Haixi Zhang, Tao Qin, Juan Zhao","doi":"10.1186/s12896-024-00879-w","DOIUrl":"https://doi.org/10.1186/s12896-024-00879-w","url":null,"abstract":"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.","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"23 1","pages":""},"PeriodicalIF":3.5,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141941949","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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