The legacy of Micheal Chevruel’s discovery of “cholesterine” as a non-saponifiable lipid from gall stones has ignited the imagination and research of countless minds for over two centuries now. In this review, we have provided a brief chronicle of the early history of cholesterol research which paved the way to present day understanding of membrane biology. We have discussed the properties and functionality of various fluorescent analogs of cholesterol in view of the ultra-high sensitivity, rapid response and spatial resolution obtained using fluorescence spectroscopic, microscopic and flow cytometric techniques. The repertoire of fluorescent analogs discussed for cholesterol research include the naturally occurring analogs (dehydroergosterol and cholestatrienol); polarity sensitive probes (NBDand dansyl-cholesterol); bright and photostable probe (BODIPY-cholesterol); clickable alkyne cholesterol and cholesterol binding macromolecules (fluorescently labeled non-toxic subunits of perfringolysin O and filipin) in monitoring cholesterol content in live and fixed cells. We have elaborated on the applications of the fluorescent analogs of cholesterol in clinical research, taking atherosclerosis, Niemann–Pick C and Alzheimer’s disease as representative examples. The applicability of fluorescent probes of cholesterol has become more relevant with the advent of various super-resolution microscopic techniques today and holds the promise of shedding light into the molecular orchestra of lipid-protein interaction with nanometer-scale resolution.
michael Chevruel发现“胆固醇”是一种来自胆结石的非皂化脂质，这一发现的遗产在两个多世纪以来激发了无数人的想象力和研究。在这篇综述中，我们提供了胆固醇研究的早期历史的简要编年史，这为今天对膜生物学的理解铺平了道路。鉴于利用荧光光谱、显微镜和流式细胞术技术获得的超高灵敏度、快速响应和空间分辨率，我们讨论了各种胆固醇荧光类似物的性质和功能。用于胆固醇研究的荧光类似物包括天然存在的类似物(脱氢麦角甾醇和胆固醇三烯醇);极性敏感探针(nbd和丹酚-胆固醇);光稳定探针(bodipy -胆固醇);可点击炔胆固醇和胆固醇结合大分子(荧光标记的无毒亚基的perfringolysin O和filipin)在监测活细胞和固定细胞中的胆固醇含量。我们详细阐述了胆固醇荧光类似物在临床研究中的应用，以动脉粥样硬化、尼曼-匹克C和阿尔茨海默病为代表。随着各种超分辨率显微技术的出现，胆固醇荧光探针的适用性变得更加相关，并有望以纳米级分辨率揭示脂质-蛋白质相互作用的分子管弦乐队。
{"title":"Cholesterol: Revisiting its fluorescent journey on 200th anniversary of Chevruel’s “cholesterine”","authors":"Arunima Chaudhuri, Deepak Anand","doi":"10.3233/BSI-170166","DOIUrl":"https://doi.org/10.3233/BSI-170166","url":null,"abstract":"The legacy of Micheal Chevruel’s discovery of “cholesterine” as a non-saponifiable lipid from gall stones has ignited the imagination and research of countless minds for over two centuries now. In this review, we have provided a brief chronicle of the early history of cholesterol research which paved the way to present day understanding of membrane biology. We have discussed the properties and functionality of various fluorescent analogs of cholesterol in view of the ultra-high sensitivity, rapid response and spatial resolution obtained using fluorescence spectroscopic, microscopic and flow cytometric techniques. The repertoire of fluorescent analogs discussed for cholesterol research include the naturally occurring analogs (dehydroergosterol and cholestatrienol); polarity sensitive probes (NBDand dansyl-cholesterol); bright and photostable probe (BODIPY-cholesterol); clickable alkyne cholesterol and cholesterol binding macromolecules (fluorescently labeled non-toxic subunits of perfringolysin O and filipin) in monitoring cholesterol content in live and fixed cells. We have elaborated on the applications of the fluorescent analogs of cholesterol in clinical research, taking atherosclerosis, Niemann–Pick C and Alzheimer’s disease as representative examples. The applicability of fluorescent probes of cholesterol has become more relevant with the advent of various super-resolution microscopic techniques today and holds the promise of shedding light into the molecular orchestra of lipid-protein interaction with nanometer-scale resolution.","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-170166","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69857105","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}
FTIR spectroscopy is an analytical technique widely applied for studying the vibrational fingerprint of organic compounds. In recent years, it has been applied to many biomedical fields because of its potential to detect the composition and molecular structure of various biological materials without the need of probe molecules. The coupling of IR spectrometers with visible microscopes has led to perform the imaging analysis of non-homogeneous samples, such as tissues and cells, in which the biochemical and spatial information are close related. In this review, we report the most significant applications of FTIR to the study of cells in different conditions (fixed, dried and living) with the aim to monitor their biochemical modifications, either induced or naturally occurring.
{"title":"Infrared spectroscopy as a new tool for studying single living cells: Is there a niche?","authors":"S. Sabbatini, C. Conti, G. Orilisi, E. Giorgini","doi":"10.3233/BSI-170171","DOIUrl":"https://doi.org/10.3233/BSI-170171","url":null,"abstract":"FTIR spectroscopy is an analytical technique widely applied for studying the vibrational fingerprint of organic compounds. In recent years, it has been applied to many biomedical fields because of its potential to detect the composition and molecular structure of various biological materials without the need of probe molecules. The coupling of IR spectrometers with visible microscopes has led to perform the imaging analysis of non-homogeneous samples, such as tissues and cells, in which the biochemical and spatial information are close related. In this review, we report the most significant applications of FTIR to the study of cells in different conditions (fixed, dried and living) with the aim to monitor their biochemical modifications, either induced or naturally occurring.","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-170171","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69857196","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}
Previous clinical research has suggested high-affinity binding of flavopiridol (FLAP) to human blood serum proteins, specifically either human serum albumin (HSA) or human alpha-1-acid glycoprotein (hAGP), when compared to fetal bovine serum albumin (BSA) or bovine alpha-1-acid glycoprotein (bAGP) used in pre-clinical assays. This high-affinity binding was suggested as the reason for its poor human clinical trial performance as a treatment for chronic lymphocytic leukaemia (CLL). Using three biophysical techniques, specifically circular dichroism (CD), isothermal calorimetry (ITC) and fluorescence spectroscopy, I show that FLAP does not have an overly high-affinity for either fetal BSA, HSA, bAGP or hAGP. I therefore suggest an alternate hypothesis that models the albumin and alpha-1-acid glycoprotein (AGP) binding sites at the different protein concentrations used in the fetal bovine pre-clinical assay and human physiological conditions. I use analytical ultracentrifugation (AUC) experiments to determine the validity of the theoretical models. The models can also be altered to account for the elevated AGP levels and reduced albumin levels seen in human cancer patients. Major differences in the concentrations of free available FLAP are observed between the fetal bovine pre-clinical model and human physiological conditions. A number of recommendations can therefore be made on how future pre-clinical assay studies should be conducted.
{"title":"The correlation of plasma proteins binding capacity and flavopiridol cellular and clinical trial studies","authors":"Daniel P Myatt","doi":"10.3233/BSI-170165","DOIUrl":"https://doi.org/10.3233/BSI-170165","url":null,"abstract":"Previous clinical research has suggested high-affinity binding of flavopiridol (FLAP) to human blood serum proteins, specifically either human serum albumin (HSA) or human alpha-1-acid glycoprotein (hAGP), when compared to fetal bovine serum albumin (BSA) or bovine alpha-1-acid glycoprotein (bAGP) used in pre-clinical assays. This high-affinity binding was suggested as the reason for its poor human clinical trial performance as a treatment for chronic lymphocytic leukaemia (CLL). Using three biophysical techniques, specifically circular dichroism (CD), isothermal calorimetry (ITC) and fluorescence spectroscopy, I show that FLAP does not have an overly high-affinity for either fetal BSA, HSA, bAGP or hAGP. I therefore suggest an alternate hypothesis that models the albumin and alpha-1-acid glycoprotein (AGP) binding sites at the different protein concentrations used in the fetal bovine pre-clinical assay and human physiological conditions. I use analytical ultracentrifugation (AUC) experiments to determine the validity of the theoretical models. The models can also be altered to account for the elevated AGP levels and reduced albumin levels seen in human cancer patients. Major differences in the concentrations of free available FLAP are observed between the fetal bovine pre-clinical model and human physiological conditions. A number of recommendations can therefore be made on how future pre-clinical assay studies should be conducted.","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-170165","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69856990","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}
A Google search for “cholesterol” (December 2016) returned 73,300,000 hits. This is much higher compared to 9,680,000 and 14,500,000 hits for “haemoglobin” (UK spelling) and “hemoglobin” (USA spelling), respectively. There is little doubt that cholesterol is one of the most well known biochemical substances in the world. Unfortunately, to the vast majority of the people, it is associated with death due to its connection with cardiovascular disease (CVD) which is the number one cause for death globally [2]. A Google search (December 2016) for “death” and “cholesterol” returned 32,700,000 hits in contrast to 37,400,000 hits when “life” and “cholesterol” was combined. The huge number of hits linked to death is surprising, although it is understandable considering the association of cholesterol with CVD. The Google results should be treated with caution but there is no doubt that cholesterol has a negative image and it is seen by many as a molecule of disease and death, and everything possible should be done to reduce its intake through the diet. Unfortunately, this link to disease and death leads people to overlook the fact cholesterol is also essential for human life. It plays a pivotal role in the structural organisation of biological membranes, tissue repair and as a precursor to a range of hormones and synthesis of vitamin D. Cholesterol was first identified in gallstones in about 1758 by the French scientist François Poulletier de la Salle. However, it was another French scientist, Michel-Eugène Chevreul (see Fig. 1), who first named this compound as “cholesterine” at a meeting of the French Academy of Sciences on August 26, 1816 [5]. Subsequently, it was identified to be an alcohol and was called cholesterol. Chevreul was born in Angers, France in 1786 and died in Paris in 1889. He is credited with being the founding father of research on fats and oils. This year (2016) marks the 200th anniversary of the naming of this compound and to celebrate this notable occasion, I chose to produce a special issue of Biomedical Spectroscopy and Imaging devoted to cholesterol research. The issue is dedicated to the memory of my former PhD supervisor, the late Professor Dennis Chapman FRS, who was one of the first to demonstrate the role of cholesterol in modulating membrane fluidity [4,7]. It contains a number of articles from researchers who are applying diverse techniques, including spectroscopic and imaging methods, to explore the structure and function of cholesterol. Ever since it was named in 1816, numerous scientists have engaged in research to understand the structure and function of this small hydrophobic molecule. It is worthy of being described as the “the most highly decorated small molecule in biology” [3]. The scientific community have certainly appreciated the value of this molecule and 13 Nobel Prizes have been awarded to scientists who have worked on cholesterol [3]. The first of these prizes was awarded in 1928 to Windaus and Wieland who d
在谷歌搜索“胆固醇”(2016年12月)，得到7330万个结果。相比之下，“血红蛋白”(英式拼写)和“血红蛋白”(美式拼写)的点击率分别为968万和1450万。毫无疑问，胆固醇是世界上最著名的生化物质之一。不幸的是，对绝大多数人来说，它与死亡有关，因为它与心血管疾病(CVD)有关，心血管疾病是全球死亡的头号原因。2016年12月，在谷歌上搜索“死亡”和“胆固醇”，得到3270万次搜索结果，而将“生命”和“胆固醇”结合起来，得到3740万次搜索结果。尽管考虑到胆固醇与心血管疾病的关系，这是可以理解的，但与死亡相关的大量撞击令人惊讶。谷歌的结果应该谨慎对待，但毫无疑问，胆固醇有一个负面的形象，它被许多人视为疾病和死亡的分子，应该尽一切可能通过饮食减少它的摄入量。不幸的是，这种与疾病和死亡的联系使人们忽视了胆固醇对人类生命也是必不可少的这一事实。它在生物膜的结构组织、组织修复和一系列激素的前体以及维生素d的合成中起着关键作用。大约在1758年，法国科学家franois Poulletier de la Salle首次在胆结石中发现了胆固醇。然而，在1816年8月26日法国科学院的一次会议上，另一位法国科学家michel - eug Chevreul(见图1)首次将这种化合物命名为“胆固醇”。后来，它被鉴定为一种酒精，并被称为胆固醇。谢弗勒尔1786年出生于法国昂热，1889年在巴黎去世。他被认为是脂肪和油脂研究的奠基人。今年(2016年)是这种化合物命名200周年，为了庆祝这一值得纪念的时刻，我选择了专门研究胆固醇的《生物医学光谱与成像》特刊。本期特为纪念我的前博士导师，已故的Dennis Chapman FRS教授，他是最早证明胆固醇在调节膜流动性中的作用的人之一[4,7]。它包含了许多研究人员的文章，他们正在应用不同的技术，包括光谱和成像方法，来探索胆固醇的结构和功能。自从它在1816年被命名以来，许多科学家从事研究，以了解这种小疏水分子的结构和功能。它不愧是“生物学中装饰最精美的小分子”。科学界当然很欣赏这种分子的价值，13位从事胆固醇研究的科学家获得了诺贝尔奖。1928年，温道斯和维兰首次获得这一奖项，他们确定了胆固醇的结构。最近的奖项颁给了布朗和戈尔茨坦，以表彰他们在胆固醇代谢调节方面的发现。在最近的一篇文章中，Brown和Goldstein以以下方式强调了胆固醇在心血管疾病中的作用:“在工业化国家，四分之一的死亡是由冠心病造成的。一个世纪的研究揭示了主要的致病因子:携带胆固醇的低密度脂蛋白
{"title":"Cholesterol: A chemical of life and death.","authors":"P. Haris","doi":"10.3233/BSI-160161","DOIUrl":"https://doi.org/10.3233/BSI-160161","url":null,"abstract":"A Google search for “cholesterol” (December 2016) returned 73,300,000 hits. This is much higher compared to 9,680,000 and 14,500,000 hits for “haemoglobin” (UK spelling) and “hemoglobin” (USA spelling), respectively. There is little doubt that cholesterol is one of the most well known biochemical substances in the world. Unfortunately, to the vast majority of the people, it is associated with death due to its connection with cardiovascular disease (CVD) which is the number one cause for death globally [2]. A Google search (December 2016) for “death” and “cholesterol” returned 32,700,000 hits in contrast to 37,400,000 hits when “life” and “cholesterol” was combined. The huge number of hits linked to death is surprising, although it is understandable considering the association of cholesterol with CVD. The Google results should be treated with caution but there is no doubt that cholesterol has a negative image and it is seen by many as a molecule of disease and death, and everything possible should be done to reduce its intake through the diet. Unfortunately, this link to disease and death leads people to overlook the fact cholesterol is also essential for human life. It plays a pivotal role in the structural organisation of biological membranes, tissue repair and as a precursor to a range of hormones and synthesis of vitamin D. Cholesterol was first identified in gallstones in about 1758 by the French scientist François Poulletier de la Salle. However, it was another French scientist, Michel-Eugène Chevreul (see Fig. 1), who first named this compound as “cholesterine” at a meeting of the French Academy of Sciences on August 26, 1816 [5]. Subsequently, it was identified to be an alcohol and was called cholesterol. Chevreul was born in Angers, France in 1786 and died in Paris in 1889. He is credited with being the founding father of research on fats and oils. This year (2016) marks the 200th anniversary of the naming of this compound and to celebrate this notable occasion, I chose to produce a special issue of Biomedical Spectroscopy and Imaging devoted to cholesterol research. The issue is dedicated to the memory of my former PhD supervisor, the late Professor Dennis Chapman FRS, who was one of the first to demonstrate the role of cholesterol in modulating membrane fluidity [4,7]. It contains a number of articles from researchers who are applying diverse techniques, including spectroscopic and imaging methods, to explore the structure and function of cholesterol. Ever since it was named in 1816, numerous scientists have engaged in research to understand the structure and function of this small hydrophobic molecule. It is worthy of being described as the “the most highly decorated small molecule in biology” [3]. The scientific community have certainly appreciated the value of this molecule and 13 Nobel Prizes have been awarded to scientists who have worked on cholesterol [3]. The first of these prizes was awarded in 1928 to Windaus and Wieland who d","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-160161","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69857294","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}
Kenneth J. Rothschild’s (see Fig. 1) interest in science and specifically spectroscopy began at an early age. In my personal communication with Ken, he told me that as a high school student (1961– 1965), at Raritan High School (Hazlet, NJ, USA), he built a spectroscopic device for identifying different compounds. Between 1965–1969, Ken was an undergraduate student at Rensselaer Polytechnic Institute (RPI) where he majored in Physics. During this period he had some notable achievements. This included
罗斯柴尔德(见图1)对科学，特别是光谱学的兴趣在很小的时候就开始了。在我与Ken的个人交流中，他告诉我，作为一名高中生(1961 - 1965)，他在美国新泽西州哈兹莱特的黎登高中(haritan high school)建造了一个光谱装置来识别不同的化合物。1965年至1969年间，肯是伦斯勒理工学院(RPI)的一名本科生，主修物理学。在此期间，他取得了一些显著的成就。这包括
{"title":"Kenneth J. Rothschild - A pioneer of infrared difference spectroscopy of membrane proteins","authors":"P. Haris","doi":"10.3233/BSI-160150","DOIUrl":"https://doi.org/10.3233/BSI-160150","url":null,"abstract":"Kenneth J. Rothschild’s (see Fig. 1) interest in science and specifically spectroscopy began at an early age. In my personal communication with Ken, he told me that as a high school student (1961– 1965), at Raritan High School (Hazlet, NJ, USA), he built a spectroscopic device for identifying different compounds. Between 1965–1969, Ken was an undergraduate student at Rensselaer Polytechnic Institute (RPI) where he majored in Physics. During this period he had some notable achievements. This included","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-160150","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69856555","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}
The 16th European Conference on Spectroscopy of Biological Molecules (ECSBM) was held at Ruhr University Bochum in Germany (6–10 September, 2015). The conference was chaired by Klaus Gerwert (see Fig. 1) and it coincided with the 30th year of this conference series. The conference is held in alternate years in different countries within Europe. The first ECSBM was held at the University of Reims in September 1985. It was organised by Lucien Bernard (Rector Reims University) and his colleagues Alain Alix and Michel Manfait. In total three out of the 16th ECSBM conferences has been held in Germany and this reflects the large number of German scientists and institutions that focus on application of spectroscopy in biological research. The second ECSBM meeting was held in Freiburg, Germany, in September 1987 and after 18 years, in 2005, the conference returned back to Germany once more. The meeting was held than in Aschaffenburg. It is noteworthy that the 16th ECSBM conference in Bochum was a form of reunion for key scientists who were responsible for starting this conference series including Prof. Werner Mäntele (see Fig. 1). Werner, who is currently based at the Johann Wolfgang Goethe University Frankfurt am Main, was awarded the Life Time Achievement Award at the Bochum meeting for his outstanding contributions in the field of biological vibrational spectroscopy. The conference in Bochum now (see Fig. 1) had a total of 222 participants from 26 different countries covering the continents of Europe, Asia and America. There were 69 lectures (15 invited lectures, 18 contributed talks and 36 selected talks from submitted abstracts). This special issue of Biomedical Spectroscopy and Imaging presents a selection of work presented at the conference. It provides excellent examples of application of spectroscopic and imaging techniques for the study of diverse biological systems. The conference in Bochum was dominated by impressive presentations which show how vibrational spectroscopy is used at different scales to solve biological questions. The talks demonstrated in a convincing manner how the use of time-resolved FTIR difference spectroscopy is able to determine molecular reaction mechanisms of proteins and protein interactions at the atomic level. For example the molecular reaction mechanisms of retinal-proteins like bacteriorhodopsin and channelrhodopsins and
{"title":"Thirty years of European Conference on Spectroscopy of Biological Molecules celebrated in Ruhr University Bochum","authors":"P. Haris, K. Gerwert","doi":"10.3233/BSI-160142","DOIUrl":"https://doi.org/10.3233/BSI-160142","url":null,"abstract":"The 16th European Conference on Spectroscopy of Biological Molecules (ECSBM) was held at Ruhr University Bochum in Germany (6–10 September, 2015). The conference was chaired by Klaus Gerwert (see Fig. 1) and it coincided with the 30th year of this conference series. The conference is held in alternate years in different countries within Europe. The first ECSBM was held at the University of Reims in September 1985. It was organised by Lucien Bernard (Rector Reims University) and his colleagues Alain Alix and Michel Manfait. In total three out of the 16th ECSBM conferences has been held in Germany and this reflects the large number of German scientists and institutions that focus on application of spectroscopy in biological research. The second ECSBM meeting was held in Freiburg, Germany, in September 1987 and after 18 years, in 2005, the conference returned back to Germany once more. The meeting was held than in Aschaffenburg. It is noteworthy that the 16th ECSBM conference in Bochum was a form of reunion for key scientists who were responsible for starting this conference series including Prof. Werner Mäntele (see Fig. 1). Werner, who is currently based at the Johann Wolfgang Goethe University Frankfurt am Main, was awarded the Life Time Achievement Award at the Bochum meeting for his outstanding contributions in the field of biological vibrational spectroscopy. The conference in Bochum now (see Fig. 1) had a total of 222 participants from 26 different countries covering the continents of Europe, Asia and America. There were 69 lectures (15 invited lectures, 18 contributed talks and 36 selected talks from submitted abstracts). This special issue of Biomedical Spectroscopy and Imaging presents a selection of work presented at the conference. It provides excellent examples of application of spectroscopic and imaging techniques for the study of diverse biological systems. The conference in Bochum was dominated by impressive presentations which show how vibrational spectroscopy is used at different scales to solve biological questions. The talks demonstrated in a convincing manner how the use of time-resolved FTIR difference spectroscopy is able to determine molecular reaction mechanisms of proteins and protein interactions at the atomic level. For example the molecular reaction mechanisms of retinal-proteins like bacteriorhodopsin and channelrhodopsins and","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-160142","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69856545","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}
A. Aluigi, G. Sotgiu, A. Torreggiani, R. Zamboni, A. Guerrini, G. Varchi, V. Orlandi
BACKGROUND: The design of wound dressings with extraordinary functionalities that fully address the problem of wound healing is an ambitious challenge in biomedical field. Keratin is a protein most abundant in nature, being the major component of wool, feather, hair, etc., with promising applications in biomedical and regenerative medicine fields. A high level of antibacterial functionality is another desirable property for applications in biomedical field in response to the increasing resistance of bacteria to antibiotics. One of the emerging methods of disinfection and sterilization is the antimicrobial photodynamic therapy (APDT), which uses light combined to a photosensitizer and oxygen to produce phototoxic species. OBJECTIVE: Biomatrices (photo-active keratin) made of wool keratin functionalized with methylene blue, a powerful photosensitizer, have been developed and tested as systems that combine the bioactive properties with the antimicrobial photodynamic functionality. METHODS: The biomatrix resistance to photo-degradation and the formation of reactive oxygen species were evaluated by spectroscopic methods, whereas the antibacterial properties were tested towards gram-positive bacteria. RESULTS: The Raman analysis revealed that specific damages occur at sensitive amino acid sites, selectively, rather than indiscriminately. However, keratin resulted to be a suitable biomaterial for APDT, since it has enough resistance to photodegradation and the radical-induced oxidation is not able to induce strong structural changes in the protein. CONCLUSIONS: The results clearly indicate the potential use of these novel photo-active keratin biomatrices in wound dressing and tissue engineering.
{"title":"Raman spectroscopic characterisation of photo-active keratin doped with Methylene Blue for wound dressings and tissue engineering","authors":"A. Aluigi, G. Sotgiu, A. Torreggiani, R. Zamboni, A. Guerrini, G. Varchi, V. Orlandi","doi":"10.3233/BSI-160143","DOIUrl":"https://doi.org/10.3233/BSI-160143","url":null,"abstract":"BACKGROUND: The design of wound dressings with extraordinary functionalities that fully address the problem of wound healing is an ambitious challenge in biomedical field. Keratin is a protein most abundant in nature, being the major component of wool, feather, hair, etc., with promising applications in biomedical and regenerative medicine fields. A high level of antibacterial functionality is another desirable property for applications in biomedical field in response to the increasing resistance of bacteria to antibiotics. One of the emerging methods of disinfection and sterilization is the antimicrobial photodynamic therapy (APDT), which uses light combined to a photosensitizer and oxygen to produce phototoxic species. OBJECTIVE: Biomatrices (photo-active keratin) made of wool keratin functionalized with methylene blue, a powerful photosensitizer, have been developed and tested as systems that combine the bioactive properties with the antimicrobial photodynamic functionality. METHODS: The biomatrix resistance to photo-degradation and the formation of reactive oxygen species were evaluated by spectroscopic methods, whereas the antibacterial properties were tested towards gram-positive bacteria. RESULTS: The Raman analysis revealed that specific damages occur at sensitive amino acid sites, selectively, rather than indiscriminately. However, keratin resulted to be a suitable biomaterial for APDT, since it has enough resistance to photodegradation and the radical-induced oxidation is not able to induce strong structural changes in the protein. CONCLUSIONS: The results clearly indicate the potential use of these novel photo-active keratin biomatrices in wound dressing and tissue engineering.","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-160143","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69856578","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}
Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), the primary receptor for ox-LDL in endothelial cells, is a multi-ligand scavenger receptor that plays a crucial role in the pathogenesis of atherosclerosis and cardiovascular disorders and recently identified as a tumor marker. LOX-1 is naturally present in caveolae/lipid rafts in plasma membranes and disruption of these membrane domains by cholesterol-lowering drugs leads to a spatial disorganization of LOX-1 and a marked loss of specific LOX-1 function in terms of ox-LDL binding and internalization. Moreover, cholesterol depletion triggers the release of LOX-1 in exosomes and enhances shedding of LOX-1 ectodomain. We here provide an overview of the involvement of membrane and circulating cholesterol in LOX-1 function and shedding and its impact on cardiovascular pathologies and cancer. In particular, we consider the available biological and molecular evidence indicating LOX-1 as a potential therapeutic target for atherosclerosis, inflammation processes, myocardial infarction and cholesterol-lowering drugs as specific inhibitors of LOX-1 function.
{"title":"Cholesterol level regulates lectin-like oxidized low-density lipoprotein receptor-1 function","authors":"Sofia Raniolo, Giulia Vindigni, S. Biocca","doi":"10.3233/BSI-160156","DOIUrl":"https://doi.org/10.3233/BSI-160156","url":null,"abstract":"Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), the primary receptor for ox-LDL in endothelial cells, is a multi-ligand scavenger receptor that plays a crucial role in the pathogenesis of atherosclerosis and cardiovascular disorders and recently identified as a tumor marker. LOX-1 is naturally present in caveolae/lipid rafts in plasma membranes and disruption of these membrane domains by cholesterol-lowering drugs leads to a spatial disorganization of LOX-1 and a marked loss of specific LOX-1 function in terms of ox-LDL binding and internalization. Moreover, cholesterol depletion triggers the release of LOX-1 in exosomes and enhances shedding of LOX-1 ectodomain. We here provide an overview of the involvement of membrane and circulating cholesterol in LOX-1 function and shedding and its impact on cardiovascular pathologies and cancer. In particular, we consider the available biological and molecular evidence indicating LOX-1 as a potential therapeutic target for atherosclerosis, inflammation processes, myocardial infarction and cholesterol-lowering drugs as specific inhibitors of LOX-1 function.","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-160156","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69856607","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}
Cholesterol is an important component of cell plasma membrane. Due to its chemical composition (long rigid hydrophobic chain and a small polar hydroxyl group), it fits most of its structure into the lipid bilayer, where its steroid rings are in close proximity and attracted to the hydrocarbon chains of neighboring lipids. This gives a condensing effect on the packing of lipids in cell membranes creating cholesterol-enriched regions called membrane rafts, which also congregate a lot of specific proteins. Membrane rafts have been shown to work as platforms involved with signaling in diverse cellular processes, such as immune regulation, cell cycle control, membrane trafficking and fusion events. A series of studies in the last two decades have linked many of these functions with the effects of membrane cholesterol content and rafts integrity on actin cytoskeleton organization, as well as its consequences in cellular biomechanics. This was possible by using microscopy techniques before and after manipulation of cholesterol content from cell plasma membrane, using agents that are able to sequester these molecules, such as cyclodextrins. In this review we’ll give a personal perspective on these studies and how microscopy techniques were important to unravel the effects of cholesterol on actin and cellular mechanics. We will also discuss how actin and cholesterol contributes to control cell secretion and vesicular trafficking.
{"title":"Understanding the role of cholesterol in cellular biomechanics and regulation of vesicular trafficking: The power of imaging","authors":"L. O. Andrade","doi":"10.3233/BSI-160157","DOIUrl":"https://doi.org/10.3233/BSI-160157","url":null,"abstract":"Cholesterol is an important component of cell plasma membrane. Due to its chemical composition (long rigid hydrophobic chain and a small polar hydroxyl group), it fits most of its structure into the lipid bilayer, where its steroid rings are in close proximity and attracted to the hydrocarbon chains of neighboring lipids. This gives a condensing effect on the packing of lipids in cell membranes creating cholesterol-enriched regions called membrane rafts, which also congregate a lot of specific proteins. Membrane rafts have been shown to work as platforms involved with signaling in diverse cellular processes, such as immune regulation, cell cycle control, membrane trafficking and fusion events. A series of studies in the last two decades have linked many of these functions with the effects of membrane cholesterol content and rafts integrity on actin cytoskeleton organization, as well as its consequences in cellular biomechanics. This was possible by using microscopy techniques before and after manipulation of cholesterol content from cell plasma membrane, using agents that are able to sequester these molecules, such as cyclodextrins. In this review we’ll give a personal perspective on these studies and how microscopy techniques were important to unravel the effects of cholesterol on actin and cellular mechanics. We will also discuss how actin and cholesterol contributes to control cell secretion and vesicular trafficking.","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-160157","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69856675","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}
R. Wu, Hai-Yuan Sun, Baoming Zhao, Geng Deng, Zhi‐Wu Yu
Structural properties of paeonol-encapsulated liposomes containing cholesterol or stigmasterol at 37°C have been investigated by synchrotron small-angle X-ray scattering (SAXS) and wide-angle X-ray scattering (WAXS) techniques. We compared the structural properties of pure dipalmitoylphosphatidylcholine (DPPC) liposomes, sterol–DPPC liposomes, and those of paeonol–sterol–DPPC liposomes at different molar ratios. Three conclusions can be drawn: First, phase separation occurs in both sterol–DPPC and paeonol–sterol–DPPC liposomes. Second, the incorporation of paeonol molecules into sterol– DPPC liposomes weakens the membrane order. Third, cholesterol has a stronger tendency to interact with DPPC as compared to its counterpart in plant, stigmasterol.
{"title":"Structural properties of paeonol encapsulated liposomes at physiological temperature: Synchrotron small-angle and wide-angle X-ray diffraction studies","authors":"R. Wu, Hai-Yuan Sun, Baoming Zhao, Geng Deng, Zhi‐Wu Yu","doi":"10.3233/BSI-160162","DOIUrl":"https://doi.org/10.3233/BSI-160162","url":null,"abstract":"Structural properties of paeonol-encapsulated liposomes containing cholesterol or stigmasterol at 37°C have been investigated by synchrotron small-angle X-ray scattering (SAXS) and wide-angle X-ray scattering (WAXS) techniques. We compared the structural properties of pure dipalmitoylphosphatidylcholine (DPPC) liposomes, sterol–DPPC liposomes, and those of paeonol–sterol–DPPC liposomes at different molar ratios. Three conclusions can be drawn: First, phase separation occurs in both sterol–DPPC and paeonol–sterol–DPPC liposomes. Second, the incorporation of paeonol molecules into sterol– DPPC liposomes weakens the membrane order. Third, cholesterol has a stronger tendency to interact with DPPC as compared to its counterpart in plant, stigmasterol.","PeriodicalId":44239,"journal":{"name":"Biomedical Spectroscopy and Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/BSI-160162","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69857361","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}
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