Pub Date : 2024-11-15DOI: 10.1016/j.bbrc.2024.151008
Yifan Zhang , Zhenzhong Bai , Kang Song , Ying Liu , Wenbin Zhang
Metabolic diseases may be prevented by reducing carbohydrate intake and replacing plant-based diets with animal-based ones low in carbohydrates but high in protein, fat, and iron. While the effects of sugars on metabolic diseases are well-known, the role of iron remains unclear. This study aimed to explore the effects of a high-fat high-iron animal diet on body metabolism in mice. Micro-PET imaging was used to assess 18-F-labelled glucose uptake in BAT, and the morphology, respiratory function, and oxidative stress of BAT mitochondria were examined. The underlying mechanisms were elucidated by analyzing the expression of UCP-1, PGC-1α and PPARα. The high-iron high-fat diet increased appetite, impaired glucose tolerance, and reduced insulin sensitivity. Additionally, the high-iron diet promoted gluconeogenesis only in the absence of high-fat levels. Both high-iron and high-fat diets suppressed BAT activity, increased mitochondrial oxidative stress, decreased mitochondrial respiratory function, and lowered thermogenic gene expression. Weight loss strategies focusing solely on reducing carbohydrates and increasing animal foods, like ketogenic diets, may have long-term detrimental effects on metabolic health. Prioritizing dietary diversity and monitoring overall caloric intake is advisable for optimal outcomes.
减少碳水化合物的摄入量，用碳水化合物含量低但蛋白质、脂肪和铁含量高的动物性膳食代替植物性膳食，可以预防代谢性疾病。糖对代谢性疾病的影响众所周知，但铁的作用仍不清楚。本研究旨在探讨高脂高铁动物饮食对小鼠体内新陈代谢的影响。研究人员利用显微 PET 成像技术评估了 BAT 的 18-F 标记葡萄糖摄取量，并检测了 BAT 线粒体的形态、呼吸功能和氧化应激。通过分析 UCP-1、PGC-1α 和 PPARα 的表达，阐明了其潜在机制。高铁高脂饮食增加了食欲，损害了葡萄糖耐量，降低了胰岛素敏感性。此外，只有在没有高脂肪的情况下，高铁饮食才会促进葡萄糖生成。高铁和高脂饮食都抑制了BAT的活性，增加了线粒体氧化应激，降低了线粒体呼吸功能，并降低了产热基因的表达。只注重减少碳水化合物和增加动物性食物的减肥策略，如生酮饮食，可能会对代谢健康产生长期不利影响。为了达到最佳效果，建议优先考虑饮食多样性并监控总热量摄入。
{"title":"High-iron diet damages brown adipose tissue mitochondria and exacerbates metabolic hazards of a high-fat diet","authors":"Yifan Zhang , Zhenzhong Bai , Kang Song , Ying Liu , Wenbin Zhang","doi":"10.1016/j.bbrc.2024.151008","DOIUrl":"10.1016/j.bbrc.2024.151008","url":null,"abstract":"<div><div>Metabolic diseases may be prevented by reducing carbohydrate intake and replacing plant-based diets with animal-based ones low in carbohydrates but high in protein, fat, and iron. While the effects of sugars on metabolic diseases are well-known, the role of iron remains unclear. This study aimed to explore the effects of a high-fat high-iron animal diet on body metabolism in mice. Micro-PET imaging was used to assess 18-F-labelled glucose uptake in BAT, and the morphology, respiratory function, and oxidative stress of BAT mitochondria were examined. The underlying mechanisms were elucidated by analyzing the expression of UCP-1, PGC-1α and PPARα. The high-iron high-fat diet increased appetite, impaired glucose tolerance, and reduced insulin sensitivity. Additionally, the high-iron diet promoted gluconeogenesis only in the absence of high-fat levels. Both high-iron and high-fat diets suppressed BAT activity, increased mitochondrial oxidative stress, decreased mitochondrial respiratory function, and lowered thermogenic gene expression. Weight loss strategies focusing solely on reducing carbohydrates and increasing animal foods, like ketogenic diets, may have long-term detrimental effects on metabolic health. Prioritizing dietary diversity and monitoring overall caloric intake is advisable for optimal outcomes.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"739 ","pages":"Article 151008"},"PeriodicalIF":2.5,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142661970","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}
Signal transducer and activator of transcription 3 (STAT3) is a multifactorial regulator involved in many biological responses. Alternative splicing of STAT3 pre-mRNA leads to an internal 50-nucleotide deletion of exon 23 selecting an alternative 3’ acceptor site, resulting in the generation of two splicing isoforms, STAT3α and STAT3β. STAT3β lacks 55 amino acid-residue transactivation domain at the C-terminal of STAT3α replacing seven unique amino acids. Although STAT3β was originally thought to be a dominant negative isoform of STAT3α, accumulating evidence have shown that STAT3β possesses both its unique functions and those that overlap with STAT3α in fundamental cellular processes. However, much remains unknown about STAT3 pre-mRNA alternative splicing in determining the balance between STAT3 isoforms. In this study, we identified cis-regulatory elements and CUGBP2/ETR-3 as a novel trans-acting factor that regulates STAT3 alternative splicing. Our findings demonstrate that STAT3 splicing can be modulated by CUGBP2 via association with UG-rich elements of intron 22, providing a novel insight into the mechanism of STAT3 alternative splicing. CUGBP2 would be a crucial molecule regulating the balance of STAT3 isoform expression, thus targeting CUGBP2 and its recognition sequences in intron 22 of STAT3 might impact on various biological processes regulated by STAT3 signaling pathway.
{"title":"RNA binding protein CUGBP2/ETR-3 regulates STAT3 alternative splicing","authors":"Miki Kise , So Masaki , Naoyuki Kataoka , Kenji Suzuki","doi":"10.1016/j.bbrc.2024.151000","DOIUrl":"10.1016/j.bbrc.2024.151000","url":null,"abstract":"<div><div>Signal transducer and activator of transcription 3 (STAT3) is a multifactorial regulator involved in many biological responses. Alternative splicing of STAT3 pre-mRNA leads to an internal 50-nucleotide deletion of exon 23 selecting an alternative 3’ acceptor site, resulting in the generation of two splicing isoforms, STAT3α and STAT3β. STAT3β lacks 55 amino acid-residue transactivation domain at the C-terminal of STAT3α replacing seven unique amino acids. Although STAT3β was originally thought to be a dominant negative isoform of STAT3α, accumulating evidence have shown that STAT3β possesses both its unique functions and those that overlap with STAT3α in fundamental cellular processes. However, much remains unknown about STAT3 pre-mRNA alternative splicing in determining the balance between STAT3 isoforms. In this study, we identified <em>cis</em>-regulatory elements and CUGBP2/ETR-3 as a novel <em>trans</em>-acting factor that regulates STAT3 alternative splicing. Our findings demonstrate that STAT3 splicing can be modulated by CUGBP2 via association with UG-rich elements of intron 22, providing a novel insight into the mechanism of STAT3 alternative splicing. CUGBP2 would be a crucial molecule regulating the balance of STAT3 isoform expression, thus targeting CUGBP2 and its recognition sequences in intron 22 of STAT3 might impact on various biological processes regulated by STAT3 signaling pathway.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"739 ","pages":"Article 151000"},"PeriodicalIF":2.5,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142646851","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}
Pub Date : 2024-11-14DOI: 10.1016/j.bbrc.2024.151003
Umate Nachiket Shankar, Sowmya Andole , Kousamvita Das , Mohd Shiraz , Mohd Akif
Many pathogens establish a successful infection by evading the host complement system, an essential arm of innate immunity. Pathogenic Leptospira is reported to escape complement-mediated killing by recruiting the host complement regulators by lipoproteins or outer surface proteins. One of the outer surface proteins, Leptospiral complement regulator-acquiring protein A (LcpA), is known to recruit complement regulators, C4b-binding protein (C4BP), and Factor H (FH) on the bacterial surface. Mapping of interacting domains from C4BP and FH with the LcpA has already been reported. However, the region or structural part of the LcpA mediating the interaction is not known yet. Here, we report cloning, expression, refolding and purification of recombinant LcpA from an inclusion body of E. coli heterologous expression system. We also demonstrate the biophysical characterization of recombinant LcpA and reveal its secondary structure contents. Moreover, the protein displays a moderate thermostability. The change of intrinsic fluorescence and CD spectra demonstrate a change in the secondary structure of protein due to binding with Zn2+ ions. Molecular docking of LcpA with the complement regulators displays important interface residues from both the individual counterparts. Molecular dynamic simulation analysis demonstrates the stability of interactions between LcpA and C4BP. In our understanding, this is the first report on the large-scale purification of LcpA through refolding experiments and biophysical characterization of LcpA. This study may provide additional information on the structural basis of binding with the complement regulators.
{"title":"Biophysical characterization and structural insights of leptospiral complement regulator-acquiring protein A","authors":"Umate Nachiket Shankar, Sowmya Andole , Kousamvita Das , Mohd Shiraz , Mohd Akif","doi":"10.1016/j.bbrc.2024.151003","DOIUrl":"10.1016/j.bbrc.2024.151003","url":null,"abstract":"<div><div>Many pathogens establish a successful infection by evading the host complement system, an essential arm of innate immunity. Pathogenic <em>Leptospira</em> is reported to escape complement-mediated killing by recruiting the host complement regulators by lipoproteins or outer surface proteins. One of the outer surface proteins, Leptospiral complement regulator-acquiring protein A (LcpA), is known to recruit complement regulators, C4b-binding protein (C4BP), and Factor H (FH) on the bacterial surface. Mapping of interacting domains from C4BP and FH with the LcpA has already been reported. However, the region or structural part of the LcpA mediating the interaction is not known yet. Here, we report cloning, expression, refolding and purification of recombinant LcpA from an inclusion body of <em>E. coli</em> heterologous expression system. We also demonstrate the biophysical characterization of recombinant LcpA and reveal its secondary structure contents. Moreover, the protein displays a moderate thermostability. The change of intrinsic fluorescence and CD spectra demonstrate a change in the secondary structure of protein due to binding with Zn<sup>2+</sup> ions. Molecular docking of LcpA with the complement regulators displays important interface residues from both the individual counterparts. Molecular dynamic simulation analysis demonstrates the stability of interactions between LcpA and C4BP. In our understanding, this is the first report on the large-scale purification of LcpA through refolding experiments and biophysical characterization of LcpA. This study may provide additional information on the structural basis of binding with the complement regulators.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"739 ","pages":"Article 151003"},"PeriodicalIF":2.5,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142661857","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}
Regulated intramembrane proteolysis (RIP) is a fundamentally conserved mechanism involving sequential cleavage by a membrane-bound Site-1 protease (S1P) and a transmembrane Site-2 protease (S2P). In the opportunistic pathogen Pseudomonas aeruginosa, the alternate sigma factor σ22 activates alginate production and in turn is regulated by the MucABCD system. The anti-sigma factor MucA, which inhibits σ22, is sequentially cleaved via RIP by AlgW (S1P) and MucP (S2P) respectively. In this study, we report high-resolution crystal structures of the MucP PDZ1 and PDZ2 domains. Structural and binding analysis confirms that MucP PDZ2 recognizes the carboxy-terminal Ala136 residue of MucA following Site-1 cleavage by AlgW, while the peptide binding groove of PDZ1 is obstructed by a short α-helix. A structure of MucP PDZ2 with bound MucA peptide shows how PDZ2 binds the newly exposed carboxyl terminus of MucA following AlgW cleavage. The ability of a ΔmucP strain of P. aeruginosa to form biofilms was reduced to a similar extent as a ΔalgW strain. This work paves the way for further studies of MucP and other PDZ-containing S2Ps in regulated intramembrane proteolysis.
{"title":"Structural insights into regulated intramembrane proteolysis by the positive alginate regulator MucP from Pseudomonas aeruginosa.","authors":"Xiaorui Lou, Shanshan Li, Yanan Wang, Runhao Wang, Weiping Li, Jiaqi Yan, Qionglin Zhang, Ruihua Liu, Mark Bartlam","doi":"10.1016/j.bbrc.2024.150999","DOIUrl":"https://doi.org/10.1016/j.bbrc.2024.150999","url":null,"abstract":"<p><p>Regulated intramembrane proteolysis (RIP) is a fundamentally conserved mechanism involving sequential cleavage by a membrane-bound Site-1 protease (S1P) and a transmembrane Site-2 protease (S2P). In the opportunistic pathogen Pseudomonas aeruginosa, the alternate sigma factor σ<sup>22</sup> activates alginate production and in turn is regulated by the MucABCD system. The anti-sigma factor MucA, which inhibits σ<sup>22</sup>, is sequentially cleaved via RIP by AlgW (S1P) and MucP (S2P) respectively. In this study, we report high-resolution crystal structures of the MucP PDZ1 and PDZ2 domains. Structural and binding analysis confirms that MucP PDZ2 recognizes the carboxy-terminal Ala136 residue of MucA following Site-1 cleavage by AlgW, while the peptide binding groove of PDZ1 is obstructed by a short α-helix. A structure of MucP PDZ2 with bound MucA peptide shows how PDZ2 binds the newly exposed carboxyl terminus of MucA following AlgW cleavage. The ability of a ΔmucP strain of P. aeruginosa to form biofilms was reduced to a similar extent as a ΔalgW strain. This work paves the way for further studies of MucP and other PDZ-containing S2Ps in regulated intramembrane proteolysis.</p>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"740 ","pages":"150999"},"PeriodicalIF":2.5,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142680703","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}
Cell migration assays, also known as chemotaxis assays, are widely used to measure the migratory capacities of cancer cells, leukocytes, macrophages, and other motile cell types. In these assays, fluorescently labeled cells are seeded onto cell culture inserts with microporous membranes that block light transmission from 490 to 700 nm. The migrated cells are then observed and quantified from the bottom of the microporous membrane using a fluorescence microscope. In this study, we conducted cell migration assays using macrophages as the motile cells. We discovered that the commonly employed fluorescent labeling method using calcein acetoxymethyl ester (calcein AM) can lead to the time-dependent attenuation of fluorescent signals in certain cell types during migration assays, potentially compromising assay stability. This study overcame this limitation by utilizing PKH26, which fluorescently labels cell surfaces through a mechanism distinct from that of calcein AM. With this modification, we observed a consistent increase in the number of migrating macrophages over time. We also demonstrated that the gradient of chemoattractants is key to the success of cell migration assays. Our improved protocol provided reliable and stable results for cell migration assays.
{"title":"Optimizing cell migration assays: Critical roles of fluorescent labeling and chemoattractant gradients","authors":"Tomomi Tadokoro , Mimoko Kato , Tatsuya Kobayashi , Hideki Taniguchi","doi":"10.1016/j.bbrc.2024.150998","DOIUrl":"10.1016/j.bbrc.2024.150998","url":null,"abstract":"<div><div>Cell migration assays, also known as chemotaxis assays, are widely used to measure the migratory capacities of cancer cells, leukocytes, macrophages, and other motile cell types. In these assays, fluorescently labeled cells are seeded onto cell culture inserts with microporous membranes that block light transmission from 490 to 700 nm. The migrated cells are then observed and quantified from the bottom of the microporous membrane using a fluorescence microscope. In this study, we conducted cell migration assays using macrophages as the motile cells. We discovered that the commonly employed fluorescent labeling method using calcein acetoxymethyl ester (calcein AM) can lead to the time-dependent attenuation of fluorescent signals in certain cell types during migration assays, potentially compromising assay stability. This study overcame this limitation by utilizing PKH26, which fluorescently labels cell surfaces through a mechanism distinct from that of calcein AM. With this modification, we observed a consistent increase in the number of migrating macrophages over time. We also demonstrated that the gradient of chemoattractants is key to the success of cell migration assays. Our improved protocol provided reliable and stable results for cell migration assays.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"739 ","pages":"Article 150998"},"PeriodicalIF":2.5,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142643830","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}
Isolated enzymes serve as advantageous platforms for the fabrication of nanomaterials. The objective of this study was to fabricate silver nanoparticles (AgNPs) incorporated with Trametes versicolor laccase and evaluate their diverse biological properties. The AgNPs fabricated through laccase-mediated methods were characterized using various characterization techniques including UV-visible (UV-vis) spectroscopy, Energy-dispersive X-ray (EDX) spectroscopy, Dynamic light scattering (DLS) spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, and Field emission scanning electron microscopy (FE-SEM). The results showed that the laccase-incorporated AgNPs were spherical in shape with a Z-average diameter of 19.40 nm and a zeta potential of -19.2 mV. The AgNPs exhibited significant dose-dependent in vitro α-amylase, urease, and DPPH free radical inhibitory activities, with maximum inhibitions of 83.49 ± 1.06 %, 68.95 ± 3.60 %, and 67.36 ± 3.40 %, respectively, at a concentration of 1000 μg mL-1. Furthermore, the intrinsic pathway-mediated anticoagulant activity of the fabricated AgNPs was confirmed through the activated partial thromboplastin time (aPTT) assay, which serves as a global coagulation assay. Additionally, the laccase-incorporated AgNPs demonstrated antibacterial properties against both standard gram-positive strains of Staphylococcus epidermidis and Streptococcus mutans, with minimum inhibitory concentration (MIC) values of 2 and 4 μg mL-1, and minimum bactericidal concentration (MBC) values of 16 and 16 μg mL-1, respectively. The dose-dependent antibacterial performance of the AgNPs against both bacterial populations was also confirmed through flow cytometry. Moreover, the AgNPs exhibited 61.53 ± 3.17 % and 63.03 ± 1.44 % biofilm degradation against S. epidermidis and S. mutans, respectively, at the maximum tested concentration (20∗MIC).
{"title":"Trametes versicolor laccase-derived silver nanoparticles: Green synthesis, structural characterization and multifunctional biological properties.","authors":"Hamed Barabadi, Melika Kamali, Kamyar Jounaki, Kimiya Karami, Salar Sadeghian-Abadi, Reza Jahani, Omid Hosseini, Salimeh Amidi","doi":"10.1016/j.bbrc.2024.150995","DOIUrl":"https://doi.org/10.1016/j.bbrc.2024.150995","url":null,"abstract":"<p><p>Isolated enzymes serve as advantageous platforms for the fabrication of nanomaterials. The objective of this study was to fabricate silver nanoparticles (AgNPs) incorporated with Trametes versicolor laccase and evaluate their diverse biological properties. The AgNPs fabricated through laccase-mediated methods were characterized using various characterization techniques including UV-visible (UV-vis) spectroscopy, Energy-dispersive X-ray (EDX) spectroscopy, Dynamic light scattering (DLS) spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, and Field emission scanning electron microscopy (FE-SEM). The results showed that the laccase-incorporated AgNPs were spherical in shape with a Z-average diameter of 19.40 nm and a zeta potential of -19.2 mV. The AgNPs exhibited significant dose-dependent in vitro α-amylase, urease, and DPPH free radical inhibitory activities, with maximum inhibitions of 83.49 ± 1.06 %, 68.95 ± 3.60 %, and 67.36 ± 3.40 %, respectively, at a concentration of 1000 μg mL<sup>-1</sup>. Furthermore, the intrinsic pathway-mediated anticoagulant activity of the fabricated AgNPs was confirmed through the activated partial thromboplastin time (aPTT) assay, which serves as a global coagulation assay. Additionally, the laccase-incorporated AgNPs demonstrated antibacterial properties against both standard gram-positive strains of Staphylococcus epidermidis and Streptococcus mutans, with minimum inhibitory concentration (MIC) values of 2 and 4 μg mL<sup>-1</sup>, and minimum bactericidal concentration (MBC) values of 16 and 16 μg mL<sup>-1</sup>, respectively. The dose-dependent antibacterial performance of the AgNPs against both bacterial populations was also confirmed through flow cytometry. Moreover, the AgNPs exhibited 61.53 ± 3.17 % and 63.03 ± 1.44 % biofilm degradation against S. epidermidis and S. mutans, respectively, at the maximum tested concentration (20∗MIC).</p>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"740 ","pages":"150995"},"PeriodicalIF":2.5,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142674921","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}
Pub Date : 2024-11-13DOI: 10.1016/j.bbrc.2024.150974
Mei-Na Li , Qiang Han , Nan Wang , Ting Wang , Xue-Ming You , Shuai Zhang , Cui-Cui Zhang , Yong-Qiang Shi , Pei-Zhuang Qiao , Cheng-Lian Man , Teng Feng , Yue-Yue Li , Zhuang Zhu , Ke-Ji Quan , Teng-Lin Xu , George Fei Zhang
16S rRNA gene sequence is the most common housekeeping genetic marker to study bacterial phylogeny and taxonomy. Therefore, 16S rRNA gene sequencing has the potential to identify novel bacteria and diagnose bacteria. This study compared 16S rRNA gene sequencing with conventional PCR for bacterial identification and disease diagnosis. The bacterial community in healthy and diseased hosts was analyzed by 16S rRNA gene sequencing. 16S rRNA gene sequencing is more sensitive than conventional PCR in detecting bacteria. Moreover, 16S rRNA gene sequencing is adequate to identify novel bacteria. 16S rRNA gene sequencing demonstrated that most pathogenic bacteria persist in diseased or healthy hosts in different abundance. Pathogenic bacteria, such as well-known chicken pathogen Avibacterium paragallinarum, Ornithobacterium rhinotracheale, and Gallibacterium anatis, were identified as indicator species of diseased samples. Alpha diversity analysis showed that the healthy group species is significantly higher than in the diseased groups. Beta diversity analysis also demonstrated differences between healthy and infected groups. The study concluded that 16S rRNA gene sequencing is a more sensitive method for detecting pathogens, and microbiota analysis can distinguish between healthy and diseased samples. Eventually, 16S rRNA gene sequencing has represented the potential in human and animal clinical diagnosis and novel bacterial identification.
{"title":"16S rRNA gene sequencing for bacterial identification and infectious disease diagnosis","authors":"Mei-Na Li , Qiang Han , Nan Wang , Ting Wang , Xue-Ming You , Shuai Zhang , Cui-Cui Zhang , Yong-Qiang Shi , Pei-Zhuang Qiao , Cheng-Lian Man , Teng Feng , Yue-Yue Li , Zhuang Zhu , Ke-Ji Quan , Teng-Lin Xu , George Fei Zhang","doi":"10.1016/j.bbrc.2024.150974","DOIUrl":"10.1016/j.bbrc.2024.150974","url":null,"abstract":"<div><div>16S rRNA gene sequence is the most common housekeeping genetic marker to study bacterial phylogeny and taxonomy. Therefore, 16S rRNA gene sequencing has the potential to identify novel bacteria and diagnose bacteria. This study compared 16S rRNA gene sequencing with conventional PCR for bacterial identification and disease diagnosis. The bacterial community in healthy and diseased hosts was analyzed by 16S rRNA gene sequencing. 16S rRNA gene sequencing is more sensitive than conventional PCR in detecting bacteria. Moreover, 16S rRNA gene sequencing is adequate to identify novel bacteria. 16S rRNA gene sequencing demonstrated that most pathogenic bacteria persist in diseased or healthy hosts in different abundance. Pathogenic bacteria, such as well-known chicken pathogen <em>Avibacterium paragallinarum</em>, <em>Ornithobacterium rhinotracheale</em>, and <em>Gallibacterium anatis</em>, were identified as indicator species of diseased samples. Alpha diversity analysis showed that the healthy group species is significantly higher than in the diseased groups. Beta diversity analysis also demonstrated differences between healthy and infected groups. The study concluded that 16S rRNA gene sequencing is a more sensitive method for detecting pathogens, and microbiota analysis can distinguish between healthy and diseased samples. Eventually, 16S rRNA gene sequencing has represented the potential in human and animal clinical diagnosis and novel bacterial identification.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"739 ","pages":"Article 150974"},"PeriodicalIF":2.5,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142646833","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}
Preeclampsia (PE) is a complex multi-organ disorder characterized by systemic inflammation, endothelial dysfunction, and vasoconstriction, which manifests as hypertension, with or without proteinuria. Effective preventive strategies for PE are currently lacking in clinical practice, leading to significant morbidity and mortality among mothers and newborns.
Objective
This study aims to investigate the impact of metformin (MET) on the l-NAME-induced PE rat model, focusing on the mechanisms through which MET may exert its effects.
Methods
Thirty pregnant Sprague-Dawley (SD) rats were randomly assigned to three groups: Control, PE, and PE + MET, on gestational day 0 (GD0). Regularly measure blood pressure and 24-h proteinuria, and collect tissue samples on GD20. Enzyme-linked immunosorbent assay (ELISA) was used to analyze inflammatory factors, endothelial function biomarkers, angiogenic factors, and apoptosis-related factors in the rat plasma. Western blot, RT-qPCR, and immunohistochemistry techniques were employed to determine the expression levels of key apoptotic proteins in placental tissue.
Results
The study findings demonstrate that MET administration improves blood pressure and 24-h proteinuria, alleviates fetal growth restriction, ameliorate inflammation cytokines, and restores the balance of angiogenic factors and endothelial function. Moreover, MET inhibits the expression levels of critical apoptotic proteins in the plasma and placental tissue of PE-like rats.
Conclusion
The results suggest that MET shows promise in alleviating symptoms associated with l-NAME-induced PE in rats, preserving endothelial function, enhancing angiogenesis, reducing inflammation, inhibiting placental apoptosis, improving placental function, and promoting fetal growth. These findings highlight MET as a potential therapeutic agent for the prevention and treatment of preeclampsia.
背景：子痫前期（PE）是一种复杂的多器官疾病，以全身炎症、内皮功能障碍和血管收缩为特征，表现为高血压，伴有或不伴有蛋白尿。目前，临床上缺乏有效的 PE 预防策略，导致母亲和新生儿的发病率和死亡率显著上升：本研究旨在探讨二甲双胍（MET）对 l-NAME 诱导的 PE 大鼠模型的影响，重点是二甲双胍发挥其作用的机制：方法：将 30 只妊娠 Sprague-Dawley (SD) 大鼠随机分为三组，分别为对照组、PE 组和 PE + MET 组：方法：30 只妊娠 Sprague-Dawley (SD) 大鼠在妊娠第 0 天（GD0）被随机分为三组：对照组、PE 组和 PE + MET 组。定期测量血压和 24 小时蛋白尿，并在 GD20 时采集组织样本。使用酶联免疫吸附试验（ELISA）分析大鼠血浆中的炎症因子、内皮功能生物标志物、血管生成因子和细胞凋亡相关因子。研究采用了 Western 印迹、RT-qPCR 和免疫组化技术来确定胎盘组织中关键凋亡蛋白的表达水平：研究结果表明，服用 MET 可改善血压和 24 小时蛋白尿，缓解胎儿生长受限，改善炎症细胞因子，恢复血管生成因子的平衡和内皮功能。此外，MET还能抑制PE样大鼠血浆和胎盘组织中关键凋亡蛋白的表达水平：结果表明，MET有望缓解l-NAME诱导的PE大鼠的相关症状，保护内皮功能，增强血管生成，减轻炎症反应，抑制胎盘凋亡，改善胎盘功能，促进胎儿生长。这些研究结果突出表明，MET 是预防和治疗子痫前期的一种潜在治疗药物。
{"title":"Protective effect of metformin on the NG-nitro-l-arginine methyl ester (l-NAME)-induced rat models of preeclampsia","authors":"Huiniu Hao, Feng Li, Fang Wang, Jia Ran, Yinmin Chen, Lijun Yang, Huijing Ma, Jianli Wang, Hailan Yang","doi":"10.1016/j.bbrc.2024.150996","DOIUrl":"10.1016/j.bbrc.2024.150996","url":null,"abstract":"<div><h3>Background</h3><div>Preeclampsia (PE) is a complex multi-organ disorder characterized by systemic inflammation, endothelial dysfunction, and vasoconstriction, which manifests as hypertension, with or without proteinuria. Effective preventive strategies for PE are currently lacking in clinical practice, leading to significant morbidity and mortality among mothers and newborns.</div></div><div><h3>Objective</h3><div>This study aims to investigate the impact of metformin (MET) on the <span>l</span>-NAME-induced PE rat model, focusing on the mechanisms through which MET may exert its effects.</div></div><div><h3>Methods</h3><div>Thirty pregnant Sprague-Dawley (SD) rats were randomly assigned to three groups: Control, PE, and PE + MET, on gestational day 0 (GD0). Regularly measure blood pressure and 24-h proteinuria, and collect tissue samples on GD20. Enzyme-linked immunosorbent assay (ELISA) was used to analyze inflammatory factors, endothelial function biomarkers, angiogenic factors, and apoptosis-related factors in the rat plasma. Western blot, RT-qPCR, and immunohistochemistry techniques were employed to determine the expression levels of key apoptotic proteins in placental tissue.</div></div><div><h3>Results</h3><div>The study findings demonstrate that MET administration improves blood pressure and 24-h proteinuria, alleviates fetal growth restriction, ameliorate inflammation cytokines, and restores the balance of angiogenic factors and endothelial function. Moreover, MET inhibits the expression levels of critical apoptotic proteins in the plasma and placental tissue of PE-like rats.</div></div><div><h3>Conclusion</h3><div>The results suggest that MET shows promise in alleviating symptoms associated with <span>l</span>-NAME-induced PE in rats, preserving endothelial function, enhancing angiogenesis, reducing inflammation, inhibiting placental apoptosis, improving placental function, and promoting fetal growth. These findings highlight MET as a potential therapeutic agent for the prevention and treatment of preeclampsia.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"739 ","pages":"Article 150996"},"PeriodicalIF":2.5,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142646847","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}
Pub Date : 2024-11-13DOI: 10.1016/j.bbrc.2024.150976
Raliat O. Abioye , Oluwasemilogo H. Adetula , Julia Diem Hum , Chibuike C. Udenigwe
Type 2 diabetes development has been associated with islet amyloid polypeptide (IAPP) fibrillation. IAPP fibrils have various deleterious effects, such as oxidative stress and disruption of cellular membrane integrity, resulting in pancreatic β-cell toxicity. Rutin, a plant polyphenol, possesses promising cytoprotective effects as a fibrillation inhibitor. Similarly, bioactive peptides have been identified as potential inhibitors to IAPP fibrillation. In this study, the effect of peptide/polyphenol mixtures consisting of rutin and each peptide, TNGQ, MANT, and YMSV, on anti-fibrillation activity and cellular response was elucidated. Results indicated a 54.7–75.1 % decrease in thioflavin T fluorescence, confirming anti-fibrillation activity. The combination decreased the average particle diameters of IAPP more than the single inhibitors, suggesting a combined effect of peptide/rutin mixtures in enhancing anti-fibrillation activity. IAPP fibrillation-induced rat insulinoma RIN-m cell death was minimized in the presence of the peptide/rutin mixture, but the activity was lower relative to rutin alone, suggesting a non-additive effect of the mixtures. Transmission electron microscopy showed a near-complete inhibition of IAPP fibrillation by TNGQ/rutin mixtures, which translated to a decreased production of membrane-bound IAPP oligomers in RIN-m cells based on immunofluorescence staining. Additionally, TNGQ/rutin mixtures significantly decreased reactive oxygen species production by 30 %, higher than the effects of single inhibitors, but no effect was observed on glucose-stimulated insulin secretion. The results demonstrate the potential of multifunctional compounds as dual inhibitor systems in controlling IAPP fibrillation and provide insight into the implications of peptide/polyphenol mixtures towards the rational development of novel anti-diabetic nutraceutical combinations.
{"title":"Influence of anti-fibrillation TNGQ peptide and rutin combination on β-cell cytoprotective effects against IAPP-induced cell death and oxidative stress","authors":"Raliat O. Abioye , Oluwasemilogo H. Adetula , Julia Diem Hum , Chibuike C. Udenigwe","doi":"10.1016/j.bbrc.2024.150976","DOIUrl":"10.1016/j.bbrc.2024.150976","url":null,"abstract":"<div><div>Type 2 diabetes development has been associated with islet amyloid polypeptide (IAPP) fibrillation. IAPP fibrils have various deleterious effects, such as oxidative stress and disruption of cellular membrane integrity, resulting in pancreatic β-cell toxicity. Rutin, a plant polyphenol, possesses promising cytoprotective effects as a fibrillation inhibitor. Similarly, bioactive peptides have been identified as potential inhibitors to IAPP fibrillation. In this study, the effect of peptide/polyphenol mixtures consisting of rutin and each peptide, TNGQ, MANT, and YMSV, on anti-fibrillation activity and cellular response was elucidated. Results indicated a 54.7–75.1 % decrease in thioflavin T fluorescence, confirming anti-fibrillation activity. The combination decreased the average particle diameters of IAPP more than the single inhibitors, suggesting a combined effect of peptide/rutin mixtures in enhancing anti-fibrillation activity. IAPP fibrillation-induced rat insulinoma RIN-m cell death was minimized in the presence of the peptide/rutin mixture, but the activity was lower relative to rutin alone, suggesting a non-additive effect of the mixtures. Transmission electron microscopy showed a near-complete inhibition of IAPP fibrillation by TNGQ/rutin mixtures, which translated to a decreased production of membrane-bound IAPP oligomers in RIN-m cells based on immunofluorescence staining. Additionally, TNGQ/rutin mixtures significantly decreased reactive oxygen species production by 30 %, higher than the effects of single inhibitors, but no effect was observed on glucose-stimulated insulin secretion. The results demonstrate the potential of multifunctional compounds as dual inhibitor systems in controlling IAPP fibrillation and provide insight into the implications of peptide/polyphenol mixtures towards the rational development of novel anti-diabetic nutraceutical combinations.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"739 ","pages":"Article 150976"},"PeriodicalIF":2.5,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142661856","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}
Pub Date : 2024-11-13DOI: 10.1016/j.bbrc.2024.150994
Shasha Liang , Yifei Qian , Ying Liu , Yahui Wang , Lianlin Su , Shuai Yan
Following abdominal surgery, the occurrence of postoperative abdominal adhesion (PAA) is highly prevalent and stands out as one of the most frequently encountered complications. The effect and molecular mechanisms of Ligustrazine nanoparticles (LN) underlying epithelial-mesenchymal transition (EMT) in PAA still remain elusive.
Adhesions were induced in Male Sprague-Dawley rats by injuring the cecum (cecal abrasion model), followed by administration of LN and hyaluronate acid (HA). The mechanism was further verified by enzyme-linked immunosorbent assay, wound healing assay, si-RNA and Western blot. Animal experiments revealed that LN effectively ameliorated adhesions, notably decreased tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-8, and fibrosis, and reduced the expression of TGF-β1 and EMT related markers (Fibronectin and E-cadherin). Furthermore, in vitro experiments demonstrated that LN might inhibit the TGF-β1 FOXC2 pathway through suppressing the expression of Fibronectin, P120, and E-cadherin and ameliorating peritoneal adhesion. Collectively, our findings indicate that LN inhibits PAA formation by reducing inflammation, decreasing EMT and promoting peritoneal mesothelial cell repair. Therefore, LN might be considered a potential candidate for the treatment of PPA. However, further clinical studies are required to approve the effectiveness of LN.
{"title":"Ligustrazine nanoparticles inhibits epithelial-mesenchymal transition and alleviates postoperative abdominal adhesion","authors":"Shasha Liang , Yifei Qian , Ying Liu , Yahui Wang , Lianlin Su , Shuai Yan","doi":"10.1016/j.bbrc.2024.150994","DOIUrl":"10.1016/j.bbrc.2024.150994","url":null,"abstract":"<div><div>Following abdominal surgery, the occurrence of postoperative abdominal adhesion (PAA) is highly prevalent and stands out as one of the most frequently encountered complications. The effect and molecular mechanisms of Ligustrazine nanoparticles (LN) underlying epithelial-mesenchymal transition (EMT) in PAA still remain elusive.</div><div>Adhesions were induced in Male Sprague-Dawley rats by injuring the cecum (cecal abrasion model), followed by administration of LN and hyaluronate acid (HA). The mechanism was further verified by enzyme-linked immunosorbent assay, wound healing assay, si-RNA and Western blot. Animal experiments revealed that LN effectively ameliorated adhesions, notably decreased tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-8, and fibrosis, and reduced the expression of TGF-β1 and EMT related markers (Fibronectin and E-cadherin). Furthermore, in vitro experiments demonstrated that LN might inhibit the TGF-β1 FOXC2 pathway through suppressing the expression of Fibronectin, P120, and E-cadherin and ameliorating peritoneal adhesion. Collectively, our findings indicate that LN inhibits PAA formation by reducing inflammation, decreasing EMT and promoting peritoneal mesothelial cell repair. Therefore, LN might be considered a potential candidate for the treatment of PPA. However, further clinical studies are required to approve the effectiveness of LN.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"739 ","pages":"Article 150994"},"PeriodicalIF":2.5,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142638327","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}
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