Integrative biology : quantitative biosciences from nano to macro最新文献

{"title":"RhoA-induced cytoskeletal tension controls adaptive cellular remodeling to mechanical signaling.","authors":"Soon-Mi Lim,&nbsp;Jerome P Trzeciakowski,&nbsp;Harini Sreenivasappa,&nbsp;Lawrence J Dangott,&nbsp;Andreea Trache","doi":"10.1039/c2ib20008b","DOIUrl":"https://doi.org/10.1039/c2ib20008b","url":null,"abstract":"<p><p>The ability to measure real-time mechanosensitive events at the subcellular level in response to discrete mechanical stimulation is a critical component in understanding mechanically-induced cellular remodeling. Vascular smooth muscle cells (VSMC) were transfected with RhoA constructs (wild type, dominant negative or constitutively active) or treated with ML-7 to induce specific cytoskeletal tension characteristics prior to mechanical stimulation. Tensile stress was applied to live VSMC using an atomic force microscope probe functionalized with extracellular matrix (ECM) proteins. The ECM induces selective integrin activation and focal adhesion formation, enabling direct manipulation of cortical actin through an active ECM-integrin-actin linkage. Therefore, locally induced mechanosensitive events triggered downstream activation of intracellular signaling pathways responsible for actin and focal adhesion remodeling throughout the cell. Integration of mechanical stimulation with simultaneous fluorescence imaging by spinning-disk confocal and total internal reflection fluorescence microscopy enabled visualization and quantification of molecular dynamic events at the sub-cellular level in real-time. Results provide evidence that the pre-existing cytoskeletal tension affects the actomyosin apparatus which in turn coordinates the ability of the cell to adapt to the externally applied stress. RhoA activation induced high cytoskeletal tension that correlated with increased stress fiber formation, cell stiffness, integrin activation and myosin phosphorylation. In contrast, blocking Rho-kinase or myosin function was characterized by low cytoskeletal tension with a decreased level of stress fiber formation, lower cell stiffness and integrin activation. Our findings show that VSMC sense and adapt to physical microenvironmental changes by a coordinated response of the actomyosin apparatus necessary to establish a new homeostatic state.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"615-27"},"PeriodicalIF":2.5,"publicationDate":"2012-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1039/c2ib20008b","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40550157","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A plant virus substrate induces early upregulation of BMP2 for rapid bone formation.","authors":"Pongkwan Sitasuwan,&nbsp;L Andrew Lee,&nbsp;Peng Bo,&nbsp;Erin N Davis,&nbsp;Yuan Lin,&nbsp;Qian Wang","doi":"10.1039/c2ib20041d","DOIUrl":"https://doi.org/10.1039/c2ib20041d","url":null,"abstract":"<p><p>Many nanoscale materials have been developed to investigate the effects on stem cell differentiations via topographical and chemical cues for applications in tissue engineering and regenerative medicine. The use of plant viruses as cell supporting substrates has been of particular interest due to the rapid induction of bone marrow derived mesenchymal stem cells (BMSCs) towards osteogenic cells. In this study, the role of Tobacco mosaic virus (TMV) and its early effects on osteoinduction with particular emphasis on the regulation of bone morphogenetic protein-2 (BMP2) was examined. We observed that the cells on the virus substrate immediately aggregated and formed bone-like nodules within 24 hours. An immediate increase in BMP2 gene and protein expression for cells on the TMV substrate was observed within 8 hours of osteoinduction. Moreover, BMP2 expression was highly localized to cells within the cell aggregates. This enhanced differentiation only occurred when TMV was coated on a solid support but not upon adding the virus to the media solution. Taken together, the results from this study highlight the potential of virus-based nanomaterials to promote endogenous BMP2 production which may prove to be a unique approach to studying the regulatory mechanisms involved in early osteoblastic differentiation.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"651-60"},"PeriodicalIF":2.5,"publicationDate":"2012-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1039/c2ib20041d","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40181618","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthetic curcumin analog EF31 inhibits the growth of head and neck squamous cell carcinoma xenografts.","authors":"Shijun Zhu,&nbsp;Terry W Moore,&nbsp;Xiaoqian Lin,&nbsp;Nao Morii,&nbsp;Alessandra Mancini,&nbsp;Randy B Howard,&nbsp;Deborah Culver,&nbsp;Richard F Arrendale,&nbsp;Prabhakar Reddy,&nbsp;Taylor J Evers,&nbsp;Hongzheng Zhang,&nbsp;Gabriel Sica,&nbsp;Zhuo G Chen,&nbsp;Aiming Sun,&nbsp;Haian Fu,&nbsp;Fadlo R Khuri,&nbsp;Dong M Shin,&nbsp;James P Snyder,&nbsp;Mamoru Shoji","doi":"10.1039/c2ib20007d","DOIUrl":"https://doi.org/10.1039/c2ib20007d","url":null,"abstract":"<p><p>Objectives are to examine the efficacy, pharmacokinetics, and toxicology of a synthetic curcumin analog EF31 in head and neck squamous cell carcinoma. The synthesis of EF31 was described for the first time. Solubility of EF24 and EF31 was compared using nephelometric analysis. Human head and neck squamous cell carcinoma Tu212 xenograft tumors were established in athymic nude mice and treated with EF31 i.p. once daily five days a week for about 5-6 weeks. The long term effect of EF31 on the NF-κB signaling system in the tumors was examined by Western blot analysis. EF31 at 25 mg kg(-1), i.p. inhibited tumor growth almost completely. Solubilities of EF24 and EF31 are <10 and 13 μg mL(-1) or <32 and 47 μM, respectively. The serum chemistry profiles of treated mice were within the limits of normal, they revealed a linear increase of C(max). EF31 decreased the level of phosphorylation of NF-κB p65. In conclusion, the novel synthetic curcumin analog EF31 is efficacious in inhibiting the growth of Tu212 xenograft tumors and may be useful for treating head and neck squamous cell carcinoma. The long term EF31 treatment inhibited NF-κB p65 phosphorylation in xenografts, implicating downregulation of cancer promoting transcription factors such as angiogenesis and metastasis.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"633-40"},"PeriodicalIF":2.5,"publicationDate":"2012-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1039/c2ib20007d","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40180941","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Resolving brain regions using nanostructure initiator mass spectrometry imaging of phospholipids.","authors":"Do Yup Lee,&nbsp;Virginia Platt,&nbsp;Ben Bowen,&nbsp;Katherine Louie,&nbsp;Christie A Canaria,&nbsp;Cynthia T McMurray,&nbsp;Trent Northen","doi":"10.1039/c2ib20043k","DOIUrl":"https://doi.org/10.1039/c2ib20043k","url":null,"abstract":"<p><p>In a variety of neurological diseases, pathological progression is cell type and region specific. Previous reports suggest that mass spectrometry imaging has the potential to differentiate between brain regions enriched in specific cell types. Here, we utilized a matrix-free surface mass spectrometry approach, nanostructure initiator mass spectrometry (NIMS), to show that spatial distributions of multiple lipids can be used as a 'fingerprint' to discriminate between neuronal- and glial- enriched brain regions. In addition, glial cells from different brain regions can be distinguished based on unique lipid profiles. NIMS images were generated from sagittal brain sections and were matched with immunostained serial sections to define glial cell enriched areas. Tandem mass spectrometry (LC-MS/MS QTOF) on whole brain extracts was used to identify 18 phospholipids. Multivariate statistical analysis (Nonnegative Matrix Factorization) enhanced differentiation of brain regions and cell populations compared to single ion imaging methods. This analysis resolved brain regions that are difficult to distinguish using conventional stains but are known to have distinct physiological functions. This method accurately distinguished the frontal (or somatomotor) and dorsal (or retrosplenial) regions of the cortex from each other and from the pons region.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"693-9"},"PeriodicalIF":2.5,"publicationDate":"2012-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1039/c2ib20043k","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40190879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Surface mobility regulates skeletal stem cell differentiation.","authors":"Cristina González-García, David Moratal, Richard O C Oreffo, Matthew J Dalby, Manuel Salmerón-Sánchez","doi":"10.1039/c2ib00139j","DOIUrl":"10.1039/c2ib00139j","url":null,"abstract":"<p><p>A family of polymer substrates which consists of a vinyl backbone chain with the side groups -COO(CH(2))(x)H, with x = 1, 2, 4, was prepared. Substrates with similar chemical groups but decreasing stiffness, characterized by their elastic modulus at 37 °C, as well as surface mobility, characterized by the glass transition temperature, were obtained. We have investigated whether these subtle variations in polymer chemistry lead to alterations in fibronectin (FN) adsorption and mesenchymal stem cell response. The same FN density was adsorbed on every substrate (∼450 ng cm(-2)) although the supramolecular organization of the protein at the material interface, as obtained with AFM, was different for x = 1 and the other two surfaces (x = 2, 4). Consequently, this allows one to investigate the effect of physical properties of the matrix on stem cell differentiation after ruling out any influence of protein activity. Cell adhesion was quantified by calculating the size distribution of focal adhesions. Mesenchymal stem cell differentiation to the osteoblastic lineage was determined by quantifying protein levels for osteocalcin, osteopontin and Runx2, in the absence of any additional osteogenic soluble factors in the culture media, but as a direct effect of material properties. The findings indicate the potential to modulate skeletal progenitor cell commitment to the osteoblastic lineage through surface mobility of the underlying material surface.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"531-9"},"PeriodicalIF":0.0,"publicationDate":"2012-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40144510","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Glycation of plasma proteins in type II diabetes lowers the non-covalent interaction affinities for dietary polyphenols.","authors":"Yixie Xie,&nbsp;Jianbo Xiao,&nbsp;Guoyin Kai,&nbsp;Xiaoqing Chen","doi":"10.1039/c2ib00185c","DOIUrl":"https://doi.org/10.1039/c2ib00185c","url":null,"abstract":"<p><p>Diabetes is characterized by an elevated level of glucose in the blood. This glucose can form covalent adducts with plasma proteins through a non-enzymatic process known as glycation. It has been suggested that the increasing glycation can influence the ability of plasma proteins to bind to small molecules. Herein, the difference between healthy human plasma proteins (HPP) and type II diabetes plasma proteins (TPP) in binding small molecules was investigated. TPP showed about 1-10 times lower affinities for polyphenols than HPP. The values of lg K(a)(HPP) are positive proportional to the values of lg K(a)(TPP) with excellent linear relationship. The glycation of HPP decreased the affinities for HPP by about 1.17 to 16.6 times. The difference between HPP-polyphenol interaction and TPP-polyphenol interaction was bigger for the more lipophilic polyphenols. The affinities for TPP or HPP slightly decreased with increasing hydrogen bond donor numbers of polyphenols and hardly changed with hydrogen bond acceptor numbers.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"502-7"},"PeriodicalIF":2.5,"publicationDate":"2012-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1039/c2ib00185c","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40166217","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identifying core features of adaptive metabolic mechanisms for chronic heat stress attenuation contributing to systems robustness.","authors":"Jenny Gu,&nbsp;Katrin Weber,&nbsp;Elisabeth Klemp,&nbsp;Gidon Winters,&nbsp;Susanne U Franssen,&nbsp;Isabell Wienpahl,&nbsp;Ann-Kathrin Huylmans,&nbsp;Karsten Zecher,&nbsp;Thorsten B H Reusch,&nbsp;Erich Bornberg-Bauer,&nbsp;Andreas P M Weber","doi":"10.1039/c2ib00109h","DOIUrl":"https://doi.org/10.1039/c2ib00109h","url":null,"abstract":"<p><p>The contribution of metabolism to heat stress may play a significant role in defining robustness and recovery of systems; either by providing the energy and metabolites required for cellular homeostasis, or through the generation of protective osmolytes. However, the mechanisms by which heat stress attenuation could be adapted through metabolic processes as a stabilizing strategy against thermal stress are still largely unclear. We address this issue through metabolomic and transcriptomic profiles for populations along a thermal cline where two seagrass species, Zostera marina and Zostera noltii, were found in close proximity. Significant changes captured by these profile comparisons could be detected, with a larger response magnitude observed in northern populations to heat stress. Sucrose, fructose, and myo-inositol were identified to be the most responsive of the 29 analyzed organic metabolites. Many key enzymes in the Calvin cycle, glycolysis and pentose phosphate pathways also showed significant differential expression. The reported comparison suggests that adaptive mechanisms are involved through metabolic pathways to dampen the impacts of heat stress, and interactions between the metabolome and proteome should be further investigated in systems biology to understand robust design features against abiotic stress.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"480-93"},"PeriodicalIF":2.5,"publicationDate":"2012-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1039/c2ib00109h","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40151928","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The mechanical memory of lung myofibroblasts.","authors":"Jenna L Balestrini,&nbsp;Sidharth Chaudhry,&nbsp;Vincent Sarrazy,&nbsp;Anne Koehler,&nbsp;Boris Hinz","doi":"10.1039/c2ib00149g","DOIUrl":"https://doi.org/10.1039/c2ib00149g","url":null,"abstract":"<p><p>Fibroblasts differentiate into the highly synthetic and contractile myofibroblast phenotype when exposed to substrates with an elastic modulus corresponding to pathologically stiff fibrotic tissue. Cellular responses to changes in substrate stiffness are typically analyzed after hours or days, which does not enable the monitoring of myofibroblast persistence, a hallmark of fibrosis. To determine long-lasting effects on the fibrotic behavior of lung fibroblasts, we followed a novel approach of explanting and repeatedly passaging fibroblasts on silicone substrates with stiffness representing various states of lung health. Fibrotic activity was determined by assaying for myofibroblast proliferation, cell contractility, expression of α-smooth muscle actin, extracellular matrix and active TGFβ1. As predicted, myofibroblast activity was low on healthy soft substrates and increased with increasing substrate stiffness. However, explanting and mechanically priming lung fibroblasts for 3 weeks on pathologically stiff substrates resulted in sustained myofibroblast activity even after the cells were returned to healthy soft cultures for 2 weeks. Such primed cells retained higher fibrotic activity than cells that had been exclusively cultured on soft substrates, and were not statistically different from cells continuously passaged on stiff surfaces. Inversely, priming lung fibroblasts for 3 weeks on soft substrates partially protected from myofibroblast activation after the shift to stiff substrates. Hence, mechano-sensed information relating to physical conditions of the local cellular environment could permanently induce fibrotic behavior of lung fibroblasts. This priming effect has important implications for the progression and persistence of aggressive fibrotic diseases such as idiopathic pulmonary fibrosis.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"410-21"},"PeriodicalIF":2.5,"publicationDate":"2012-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1039/c2ib00149g","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40159196","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modelling complex biological systems using an agent-based approach.","authors":"Mike Holcombe, Salem Adra, Mesude Bicak, Shawn Chin, Simon Coakley, Alison I Graham, Jeffrey Green, Chris Greenough, Duncan Jackson, Mariam Kiran, Sheila MacNeil, Afsaneh Maleki-Dizaji, Phil McMinn, Mark Pogson, Robert Poole, Eva Qwarnstrom, Francis Ratnieks, Matthew D Rolfe, Rod Smallwood, Tao Sun, David Worth","doi":"10.1039/c1ib00042j","DOIUrl":"10.1039/c1ib00042j","url":null,"abstract":"<p><p>Many of the complex systems found in biology are comprised of numerous components, where interactions between individual agents result in the emergence of structures and function, typically in a highly dynamic manner. Often these entities have limited lifetimes but their interactions both with each other and their environment can have profound biological consequences. We will demonstrate how modelling these entities, and their interactions, can lead to a new approach to experimental biology bringing new insights and a deeper understanding of biological systems.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"53-64"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40128464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Limitations of conventional inhibitor classifications.","authors":"Ryan Walsh,&nbsp;Earl Martin,&nbsp;Sultan Darvesh","doi":"10.1039/c1ib00053e","DOIUrl":"https://doi.org/10.1039/c1ib00053e","url":null,"abstract":"<p><p>Enzyme inhibitors are usually classified as competitive, non-competitive or mixed non-competitive. Each of these designations has a serious limitation in that it only describes an extreme of inhibitory behaviour. The non-competitive inhibition equation only considers an approach to complete inhibition of the catalytic turnover rate, while the competitive inhibition equation predicts an infinite increase in the Michaelis-Menten constant (decrease in enzyme affinity for substrate), resulting from increased inhibitor concentration. Both of these models exclude the possibility of a finite inhibitor-induced change in the kinetic parameters of the enzyme they are affecting. They also exclude the possibility of an inhibitor affecting both the substrate affinity and the catalytic turnover at the same time. Mixed non-competitive inhibition describes a hybrid form of inhibition displaying some characteristics of both competitive and non-competitive inhibition. It also suffers from an inability to describe finite changes in activity and to describe concomitant changes in substrate affinity and catalytic turnover. Two inhibitor binding constants are invoked in this equation, suggesting that such inhibitors interact with the enzyme in two completely independent manners. From these considerations, it is suggested here that conventional equations do not adequately describe observed kinetic data due to a lack of distinction between the mass action binding term describing inhibitor-enzyme association and the terms representing the actual effect of the inhibitor on the enzyme. Herein we describe an alternate approach for representing enzyme activity modulation based on a re-examination of conventional inhibition equations. The arguments presented are illustrated using the known competitive inhibition of Kallikrein with benzamidine.</p>","PeriodicalId":520649,"journal":{"name":"Integrative biology : quantitative biosciences from nano to macro","volume":" ","pages":"1197-201"},"PeriodicalIF":2.5,"publicationDate":"2011-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1039/c1ib00053e","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40114729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}