In one volume this book provides useful and innovative protocols developed specifically for the proteomic profiling of human tissues. The book provides high-throughput gel-based techniques, microarrays and a number of other methods used in proteomic research. This important book will prove indispensable to investigators of biomarker discovery and therapeutic response profiling, as well as those forging new paths in the fields of theranostics and personalized medicine.

A multiplicity of biotrophic micro-organisms interact with plants in nature, forming symbiotic relationships that range from mutualism to antagonism. Microorganisms that have adopted biotrophy as a lifestyle are able to colonize the plant and often to cross the plant cell boundaries by forming intracellular structures that are the site of nutrient uptake/exchange. To establish themselves within plant tissues, both mutualistic and pathogenic biotrophs need to overcome the plant defense response through an exchange of molecular signals. Our knowledge of the nature of these signals and their function in the interaction has rapidly increased over the last few years. This volume focuses on the genetic, molecular and cellular components involved in the communication between partners of well-known symbioses, but also reports on the advances for less studied systems.

DNA in the nucleus of plant and animal cells is stored in the form of chromatin. Chromatin and the Chromatin remodelling enzymes play an important role in gene transcription.

Recent major advances in our understanding of modulating protein functions has led to the development of new methods and algorithms to predict and decipher how amino acid sequences shape three-dimensional structures. Protein Design: Methods and Applications presents the most up-to-date protein design and engineering strategies so that readers can undertake their own projects with a maximum chance of success.
The authors present integrated computational approaches that require various degrees of computational complexity, and the major accomplishments that have been achieved in the design and structural characterization of helical peptides and proteins. Other topics of discussion include: design of structural elementary motifs, entire proteins, and interfaces of protein complexes, and of amyloidogenic polypeptides and amyloid inhibitors.
Authoritative and cutting-edge, Protein Design: Methods and Applications will be of major interest to protein scientists, biochemists, and all experimentalists selecting the strategy most adaptable for their design problem.

Phosphorus (P) is a finite resource which is essential for life. It is a limiting nutrient in many ecosystems but also a pollutant which can affect biodiversity in terrestrial ecosystems and change the ecology of water bodies. This book collects the latest information on biological processes in soil P cycling, which to date have remained much less understood than physico-chemical processes. The methods section presents spectroscopic techniques and the characterization of microbial P forms, as well as the use of tracers, molecular approaches and modeling of soil-plant systems. The section on processes deals with mycorrhizal symbioses, microbial P solubilization, soil macrofauna, phosphatase enzymes and rhizosphere processes. On the system level, P cycling is examined for grasslands, arctic and alpine soils, forest plantations, tropical forests, and dryland regions. Further, P management with respect to animal production and cropping, and the interactions between global change and P cycling, are treated.

The aim of this volume is to brief researchers of the importance of data analysis in enzymology, and of the modern methods that have developed concomitantly with computer hardware. It is also to validate researchers' computer programs with real and synthetic data to ascertain that the results produced are what they expected.
Selected Contents: Prediction of protein structure; modeling and studying proteins with molecular dynamics; statistical error in isothermal titration calorimetry; analysis of circular dichroism data; model comparison methods

Liposomes are cellular structures made up of lipid molecules. Important as a cellular model in the study of basic biology, liposomes are also used in clinical applications such as drug delivery and virus studies. Liposomes Part D is a continuation of previous MIE Liposome volumes A, B, and C.
Contents: Antibody or Ligand Targeted Liposomes; environment sensitive liposomes; liposomal oligonucleotides; liposomes in vivo

This volume addresses current methods in biological imaging, including extensive sections on MRI, CAT, NMR, PET and other imaging techniques.

In this volume expert researchers in the field detail the operations of microchip capillary electrophoresis. Chapters focus on small molecule, biomolecule applications, various detection modes, and sample preparation approaches are described. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Microchip Capillary Electrophoresis Protocol aids scientists in continuing to study microchip capillary electrophoresis.

Interdisciplinary knowledge is becoming more and more important to the modern scientist. This invaluable textbook covers bioanalytical chemistry (mainly the analysis of proteins and DNA) and explains everything for the nonbiologist. Electrophoresis, mass spectrometry, biosensors, bioassays, DNA and protein sequencing are not necessarily all included in conventional analytical chemistry textbooks. The book describes the basic principles and the applications of instrumental and molecular methods. It is particularly useful to chemistry and engineering students who already have some basic knowledge about analytical chemistry.

New Antisense Strategies: Chemical Synthesis of RNA Oligomers, by Junichi Yano und Gerald E. Smyth Development and Modification of Decoy Oligodeoxynucleotides for Clinical Application, by Mariana Kiomy Osako, Hironori Nakagami und Ryuichi Morishita Modulation of Endosomal Toll-Like Receptor-Mediated Immune Responses by Synthetic Oligonucleotides, by Ekambar R. Kandimalla und Sudhir Agrawal Delivery of Nucleic Acid Drugs, by Yan Lee und Kazunori Kataoka Aptamer: Biology to Applications, by Yoshikazu Nakamura Development and Clinical Applications of Nucleic Acid Therapeutics, by Veenu Aishwarya, Anna Kalota und Alan M. Gewirtz

"Immuno Systems Biology" aims to study the immune system in the more integrated manner on how cells and molecules participate at different system levels to the immune function. Through this bookKumar Selvarajoointroduces to physicists, chemists, computer scientists, biologists and immunologists the idea of an integrated approach to the understanding of mammalian immune system. Geared towards a researcher with limited immunological and computational analytical experience, the book provides a broad overview to the subject and some instruction in basic computational, theoretical and experimental approaches. The book links complex immunological processes with computational analysis and emphasizes the importance of immunology to themammalian system. "

Protein Transcription is a key element of cellular and organ regulation. This volume covers structure and function of all major elements associated with transcription.
*Mechanism of RNA polymerase I Transcription
*Structure and function of RNA Polymerase II
*Structure and function of the TFIID complex
*Functional properties of Chromatin Remodeling Enzymes
*Posttranslational modification

This book offers an overview of our current understanding of host defense peptides and their potential for clinical applications as well as some of the obstacles to this. The chapters, written by leading experts in the field, detail the number and diversity of host defense peptides, and discuss the therapeutic potential not only of antibacterial, but also of antifungal, antiviral, plant antimicrobial and anticancer host defense peptides. The authors provide new insights into their mechanisms of action and their immunomodulatory properties, and review recent advances in the design of novel therapeutic molecules. Lastly, their potential to prevent preterm births and Staphylococcus aureus infections is highlighted. The book is of interest to researchers, industry and clinicians alike.

The inducible isoforms of the enzymes cyclooxygenase (COX 2), nitric oxide synthase (iNOS) and heme oxygenase 1 (HO-1) have generated great interest as possible therapeutic targets in inflammation. This book is the first publication to address the importance of all three enzymes and the consequences of their interactions to the inflammatory process. The book brings together overviews by leading researchers in the field of the current status of knowledge of COX, NOS and HO in inflammation. These overviews cover a series of new concepts in the mechanism of inflammation. Topics include inducible enzyme involvement in inflammatory processes including the role in vascular permeability, leukocycte migration, granuloma formation, angiogenesis, neuroinflammation and algesia. New findings from transgenic animal models are reviewed. Other chapters address the importance of these enzymes in inflammatory disease states including rheumatoid arthritis, atherosclerosis and multiple sclerosis. The possibility of selective inhibitors or inducers of COX, NOS and HO, and their use in the clinic is discussed. The subject matter of this book is of interest to rheumatologists, pathologists, pharmacologists, neuroscientists and anyone with an academic interest in the mechanisms of inflammation.

Protein phosphatases are a group of enzymes responsible for the dephosphorylation of various proteins and enzymes in a cell. This role is an extremely important one since protein phosphorylation and dephosphorylation is required for the regulation of a large number of cellular activities.
* Classification of Protein Phosphatases
* Analysis/Technology: Cell and Molecular Imaging Technology, Assay of Protein Phosphatases, MS and MNR, Genomics/Proteomics, cDNA Microarray Analysis
* Cellular Regulation: Substrates, Inhibitors, Regulation, Protein-Protein Interaction
* Biological Function: Antisense Studies, Transgenic and Knockout Animal Models in Vivo, Therapeutic Approaches

This book reflects the use of cyanobacteria for the bioremediation of wastewater through different mechanisms and pathways of transformation and transfer of hazardous substances from one medium to another. The application of microorganisms for bioremediation is determined by their ubiquity, small size, high rate of reproduction and large surface-to-volume cell ratio. Mechanisms of interaction of cyanobacteria with inorganic pollutants include biosorption, bioaccumulation with an opportunity to obtain metal nanoparticles both on the cell surface and inside the cells as well as chelation and inclusion of metals in the composition of certain organic structures. Data presented in the book provides specialists in the field with useful information for bioremediation technologies as well as for obtaining valuable preparations using cyanobacteria.

The work reported in this book represents an excellent example of how creative experimentation and technology development, complemented by computational data analysis, can yield important insights that further our understanding of biological entities from a systems perspective. The book describes how the study of a single RNA-binding protein and its interaction sites led to the development of the novel 'protein occupancy profiling' technology that for the first time captured the mRNA sequence space contacted by the ensemble of expressed RNA binders. Application of protein occupancy profiling to eukaryotic cells revealed that extensive sequence stretches in 3' UTRs can be contacted by RBPs and that evolutionary conservation as well as negative selection act on protein-RNA contact sites, suggesting functional importance. Comparative analysis of the RBP-bound sequence space has the potential to unravel putative cis-acting RNA elements without a priori knowledge of the bound regulators. Here, Dr. Munschauer provides a comprehensive introduction to the field of post-transcriptional gene regulation, examines state-of-the-art technologies, and combines the conclusions from several journal articles into a coherent and logical story from the frontiers of systems-biology inspired life science. This thesis, submitted to the Department of Biology, Chemistry and Pharmacy at Freie Universitat Berlin, was selected as outstanding work by the Berlin Institute for Medical Systems Biology at the Max-Delbrueck Center for Molecular Medicine, Germany.

Liposomes are cellular structures made up of lipid molecules. Important as a cellular model in the study of basic biology, liposomes are also used in clinical applications such as drug delivery and virus studies.
* Liposomes in Biochemistry
* Liposomes in Molecular Cell Biology
* Liposomes in Molecular Virology

DNA in the nucleus of plant and animal cells is stored in the form of chromatin. Chromatin and the chromatin remodelling enzymes play an important role in gene transcription.
*Genetic assays of chromatin modification and remodeling
*Histone modifying enzymes
*ATP-dependent chromatin remodeling enzymes

This volume focuses on methods related to allosteric enzymes and receptors, including fluorescent proves, spectroscopic methods and quantitative analysis as well as on cooperativity in protein folding. NMR and mass spectrometry methods are discussed.
*Allosteric Enzymes and Receptors
*Cooperativity in Protein Folding and Assembly

Phospholipidshavelongbeenknownfortheirkeyroleinmaintainingthebilayer structureofmembranesandinphysicallyseparatingthecytosolfromorganelles andtheextracellularspace. Inthepastdecade,acompletelynovelandunexpected functionemerged,full?llingacrucialroleincellsignaling. Itwasthediscoveryin animalcells,thatagonist-activatedcellsurfacereceptorsledtotheactivationofa phospholipase C (PLC), to hydrolyze the minor lipid, phosphatidylinositol 4- bisphosphateintotwosecondmessengers,inositol1,4,5-trisphosphate(InsP)and 3 2+ diacylglycerol(DAG). WhileInsP diffusesintothecytosol,whereitreleasesCa 3 2+ from an intracellular store by activating a ligand-gated Ca -channel, DAG remainsinthemembranetorecruitandactivatemembersoftheproteinkinase Cfamily. Overtheyears,avarietyofotherlipidbased-signalingcascadesweredisc- ered. Theseinclude,phospholipaseA,generatinglyso-phospholipidsandfreefatty acids(tobeconvertedintoprostaglandinsandleukotrienes),phospholipaseD,to generatethelipidsecondmessenger,phosphatidicacid(PA),andphosphoinositide 3-kinase (PI3K), generating a distinct set of polyphosphoinositides (PPI) ph- phorylated at the D3-position of the inositol ring, all with separate signaling functions. Sphingolipids,representinganotherimportantgroupofsignalinglipids, alsocameacross. Themajorityoftheselipid-basedsignalingpathwayshavebeendiscoveredin plantcellstoo. Moreover,theyhavebeenfoundtobeactivatedinresponsetoa widevarietyofbioticandabioticstresssignals,butalsotobebasicallyinvolvedin plantgrowthanddevelopment. Whilemanyoftheenzymes,lipids,andtheirtargets involved arewell conserved, major differences with the mammalian paradigms havealsoemerged. Thisbookhighlightsthecurrentstatusofplantlipidsignaling. Allchaptershave beenwrittenbyexpertsinthe?eldandcoverinformationforbothbeginnersand advancedlipidologists. PartIincludesphospholipases(Chaps. 1-3),partII,lipid kinases (Chaps. 4-7), part III, lipid phosphatases (Chaps. 8-9), part IV, ix x Preface inositolphosphates and PPI metabolism (Chaps. 10-13), part V, PA signaling (Chaps. 14-17),andpartVI,additionallipidsignals,e. g. oxylipins,NAPEand sphingolipids(Chaps18-20). Ithasbeenagreatpleasuretobetheeditorofthis bookandtobeawitnessofthislipid-signalingadventure. Amsterdam,June2009 TeunMunnik Contents PartI Phospholipases PhospholipaseAinPlantSignalTransduction...3 Gu..ntherF. E. Scherer TheEmergingRolesofPhospholipaseCinPlantGrowth andDevelopment...23 PeterE. DowdandSimonGilroy PlantPhospholipaseD...39 WenhuaZhang,XiaoboWan,YueyunHong,WeiqiLi,andXueminWang PartII Kinases Phosphatidylinositol4-PhosphateisRequiredforTip GrowthinArabidopsisthaliana ...65 AmyL. SzumlanskiandErikNielsen PIP-KinasesasKeyRegulatorsofPlantFunction ...79 TillIschebeckandIngoHeilmann PlantPhosphatidylinositol3-Kinase...95 YureeLee,TeunMunnik,andYoungsookLee DiacylglycerolKinase...107 StevenA. AriszandTeunMunnik xi xii Contents PartIII Phosphatases SignalingandthePolyphosphoinositidePhosphatasesfromPlants ...117 GlendaE. Gillaspy PhosphatidicAcidPhosphatasesinSeedPlants...131 YukiNakamuraandHiroyukiOhta PartIV PPIMetabolism InsP inPlantCells ...145 3 YangJuIm,BrianQPhillippy,andImaraYPerera InositolPolyphosphatesandKinases...161 JillStevenson-PaulikandBrianQ. Phillippy PhosphoinositidesandPlantCellWallSynthesis ...175 RuiqinZhong,RyanL. McCarthy,andZheng-HuaYe ImagingLipidsinLivingPlants ...185 JoopE. M. VermeerandTeunMunnik PartV PASignaling PhosphatidicAcid:AnElectrostatic/Hydrogen-BondSwitch?...2 03 EdgarEduardKooijmanandChristaTesterink NitricOxideandPhosphatidicAcidSignalinginPlants...223 AyelenM. Diste'fano,M. LucianaLanteri,ArjentenHave, CarlosGarc?'a-Mata,LorenzoLamattina,andAnaM. Laxalt 3-Phosphoinositide-DependentProteinKinaseisaSwitchboard fromSignalingLipidstoProteinPhosphorylationCascades...243 ChristineZalejskiandLa'szlo'Bo..gre PartVI AdditionalLipidSignals DiacylglycerolPyrophosphate,ANovelPlantSignalingLipid...263 EmmanuelleJeannette,SophieParadis,andChristineZalejski OxylipinSignalingandPlantGrowth...277 AlinaMosblech,IvoFeussner,andIngoHeilmann Contents xiii FattyAcidAmideHydrolaseandtheMetabolismof N-AcylethanolamineLipidMediatorsinPlants...293 KentD. ChapmanandElisonB. Blanca?or SphingolipidSignalinginPlants...307 LouiseV. MichaelsonandJohnathanA. Napier Index ...323 Contributors Steven A. Arisz Section Plant Physiology, Swammerdam Institute for Life Sciences,UniversityofAmsterdam,SciencePark904,NL-1098XH,Amsterdam, TheNetherlands ElisonB. Blanca?or SamuelRobertsNobleFoundation,PlantBiologyDivision, Ardmore,OK73401,USA,eblanca?or@noble.

This manual reflects practical approaches to handling bacteria in the labora- tory. It is designed to recall historical methods of bacterial genetics that have had recent developments and to present new techniques that allow full genome analysis. It has been written for microbiologists who need to group their protocols at the state of the art of a new millennium and also for scientists in other fields of life sciences who need to use bacteria for their research. Teachers, graduate students, and postdocs also will benefit from having these protocols to help them understand modern bacterial genetics. I learned so much from these contributions from my colleagues that I have no doubt about the daily usefulness of this book. April 2002 Michel Blot XII Abbreviations Acyl-HSL N-acyl homoserine lactone moi multiplicity of infection Amp or Ap ampicillin N amino C carboxy NMR nuclear magnetic resonance CIO-HSL N-decanoyl-L-homoserine lactone 3-0H-C14:1-HSL N-(3-hydroxy-7 -cis-tetra- C12-HSL N-dodecanoyl-L-homoserine lac- decanoyl)homo-serine lactone tone 3-0H-C4-HSL N-3-hydroxybutanoyl-L- C14-HSL N-tetradecanoyl-L-homoserine homoserine lactone lactone ONPG o-nitrophenyl ~-D-galactopyranoside C4-HSL N-butanoyl-L-homoserine lactone ORF open reading frame C6-HSL N-hexanoyl-L-homoserine lactone OTG I-S-octyl-~-D-thioglucoside C8-HSL N-octanoyl-L-homoserine lactone 3-oxo-CIO-HSL N-3-oxodecanoyl-L-homo- Cam or Cm chloramphenicol serine lactone CBD chitin binding domain 3-oxo-C12-HSL N-3-oxododecanoyl-L- CHEF contour clamped homogenous electric homoserine lactone field 3-oxo-C14-HSL N-3-oxotetradecanoyl-L- CI consistency index homoserine lactone CRIM conditional-replication, integration, 3-oxo-C4-HSL N-3-oxobutanoyl-L-homoser- and modular ine lactone dCTP deoxycytidine triphosphate 3-oxo-C6-HSL N-3 -oxohexanoyl-L-homoser- deg.

Nuclear G-Protein Coupled Receptors: Methods and Protocols is a compilation of a number of conceptual and methodological aspects important for the validation and characterization of intacrine signaling systems. To date, the best-characterized intracrine signaling system is that of angiotensin II (Ang II), covered in depth in various chapters. Methodology to study the subcellular localization and function of GPCRs and other signaling systems is provided, as well as numerous chapters focusing on methods designed to understand signaling mediated by nuclear and other internal GPCRs. Methods are also described to study the formation of second messengers such as cAMP and to study the trafficking of receptors from the cell surface. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Nuclear G-Protein Coupled Receptors: Methods and Protocols seeks to serve both professionals and novices with state-of-the-art approaches to characterize what is becoming a common theme in cellular signaling.

Quinones are members of a class of aromatic compounds with two oxygen atoms bonded to the ring as carbonyl groups. This volume covers more clinical aspects of quinines, such as anticancer properties, as well as their role in nutrition and in age-related diseases.
*Mitochondrial Ubiquinone and Reductases
*Anticancer Quinones and Quinone Oxido-Reductases
*Quininone Reductases: Chemoprevention, Nutrition
*Quinones and Age-Related Diseases