Fiber sources, country and annual production of plant fibers.
\\n\\n
Released this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\\n\\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
Note: Edited in March 2021
\\n"}]',published:!0,mainMedia:{caption:"Highly Cited",originalUrl:"/media/original/117"}},components:[{type:"htmlEditorComponent",content:'IntechOpen is proud to announce that 191 of our authors have made the Clarivate™ Highly Cited Researchers List for 2020, ranking them among the top 1% most-cited.
\n\nThroughout the years, the list has named a total of 261 IntechOpen authors as Highly Cited. Of those researchers, 69 have been featured on the list multiple times.
\n\n\n\nReleased this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\n\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
Note: Edited in March 2021
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First topic will be dedicated to the principles of electrophoresis (voltage and ionic strength of buffers will also be addressed). Second topic will be on the DNA gel electrophoresis. Among other things it will address staining of gel for visualizing the DNA bands, different concentrations of agarose gel for separation of DNA of different sizes, DNA marker (used for estimation the rough molecular weight of DNA sample) etc.Third topic will be on protein gel electrophoresis. Along with other things it will address the percentages of polyacrylamide gel for separation of protein on the gel, staining of the polyacrylamide gel with Commassie Brilliant Blue G250 or R250 or silver staining. Fourth topic will cover protocol used for the working of gel electrophoresis apparatus, preparation of agarose gel and buffers. Fifth topic will cover troubleshooting with gel electrophoresis (i.e. bad resolution, heating up of apparatus, voltage problems etc.). Sixth topic will be on different companies manufacturing the gel electrophoresis apparatus.
",isbn:null,printIsbn:"979-953-307-X-X",pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,isSalesforceBook:!1,isNomenclature:!1,hash:"5ef0f1cd60f719358f76bc0ad32aa9ed",bookSignature:"Dr. Farheen Aslam",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/8173.jpg",keywords:"Voltage, Ionic Concentration, Size of Agarose Gel, Buffers, Staining of Gel, Ethidium Bromide, SDS-PAGE, Protein Marker, Agarose Gel, Positive and Negative Electrode, Bad Resolution, Voltage Problem",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"October 18th 2019",dateEndSecondStepPublish:"March 2nd 2020",dateEndThirdStepPublish:"May 1st 2020",dateEndFourthStepPublish:"July 20th 2020",dateEndFifthStepPublish:"September 18th 2020",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"2 years",secondStepPassed:!0,areRegistrationsClosed:!0,currentStepOfPublishingProcess:5,editedByType:null,kuFlag:!1,biosketch:null,coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"309320",title:"Dr.",name:"Farheen",middleName:null,surname:"Aslam",slug:"farheen-aslam",fullName:"Farheen Aslam",profilePictureURL:"https://mts.intechopen.com/storage/users/309320/images/system/309320.png",biography:"Farheen Aslam MS, PhD, is Assistant Professor at Department of Biotechnology, Lahore College for Women University, Lahore, Pakistan. She has obtained her Ph.D. from School of Biological Sciences, University of the Punjab, Pakistan in 2012: her research topic was “Cloning. expression and physicochemical analysis of proinsulin and its derivatives”. Her research field includes human biochemistry and disease, molecular biology and recombinant DNA technology, protein expression and purification, refolding of protein, enzymology, genetics and microbiology. She is proficient in use of HPLC system. MALDI-TOF, Polymerase chain reaction (PCR), Agarose Gel Electrophoresis, UV-Spectrophotometer, RNA, Plasmid and Genomic DNA Isolation. She worked as a Research Officer in the Mass Spectrometry Lab of School of Biological Sciences, University of the Punjab, Lahore from 23rd August 2010 to 23rd November 2010. Responsibilities were to prepare and analyze the samples on Voyager De Pro (ABI) MALDI TOF mass spectrometry workstation. She was awarded with a Startup Research Grant on “Studying the effect on expression of a recombinant protein by changing the nucleotides at the promoter & start of the gene” of PKR 0.5 million by Higher Education Commission on June 4th, 2013. She has participated in 19 conferences at National and international level. She has Published 11 research paper in National and International journals.",institutionString:"Lahore College for Women University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"Lahore College for Women University",institutionURL:null,country:{name:"Pakistan"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"6",title:"Biochemistry, Genetics and Molecular Biology",slug:"biochemistry-genetics-and-molecular-biology"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"184402",firstName:"Romina",lastName:"Rovan",middleName:null,title:"Ms.",imageUrl:"https://mts.intechopen.com/storage/users/184402/images/4747_n.jpg",email:"romina.r@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review, to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. Whether that be identifying an exceptional author and proposing an editorship collaboration, or contacting researchers who would like the opportunity to work with IntechOpen, I establish and help manage author and editor acquisition and contact."}},relatedBooks:[{type:"book",id:"6694",title:"New Trends in Ion Exchange Studies",subtitle:null,isOpenForSubmission:!1,hash:"3de8c8b090fd8faa7c11ec5b387c486a",slug:"new-trends-in-ion-exchange-studies",bookSignature:"Selcan Karakuş",coverURL:"https://cdn.intechopen.com/books/images_new/6694.jpg",editedByType:"Edited by",editors:[{id:"206110",title:"Dr.",name:"Selcan",surname:"Karakuş",slug:"selcan-karakus",fullName:"Selcan Karakuş"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1591",title:"Infrared Spectroscopy",subtitle:"Materials Science, Engineering and Technology",isOpenForSubmission:!1,hash:"99b4b7b71a8caeb693ed762b40b017f4",slug:"infrared-spectroscopy-materials-science-engineering-and-technology",bookSignature:"Theophile Theophanides",coverURL:"https://cdn.intechopen.com/books/images_new/1591.jpg",editedByType:"Edited by",editors:[{id:"37194",title:"Dr.",name:"Theophile",surname:"Theophanides",slug:"theophile-theophanides",fullName:"Theophile Theophanides"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3161",title:"Frontiers in Guided Wave Optics and Optoelectronics",subtitle:null,isOpenForSubmission:!1,hash:"deb44e9c99f82bbce1083abea743146c",slug:"frontiers-in-guided-wave-optics-and-optoelectronics",bookSignature:"Bishnu Pal",coverURL:"https://cdn.intechopen.com/books/images_new/3161.jpg",editedByType:"Edited by",editors:[{id:"4782",title:"Prof.",name:"Bishnu",surname:"Pal",slug:"bishnu-pal",fullName:"Bishnu Pal"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"371",title:"Abiotic Stress in Plants",subtitle:"Mechanisms and Adaptations",isOpenForSubmission:!1,hash:"588466f487e307619849d72389178a74",slug:"abiotic-stress-in-plants-mechanisms-and-adaptations",bookSignature:"Arun Shanker and B. Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3092",title:"Anopheles mosquitoes",subtitle:"New insights into malaria vectors",isOpenForSubmission:!1,hash:"c9e622485316d5e296288bf24d2b0d64",slug:"anopheles-mosquitoes-new-insights-into-malaria-vectors",bookSignature:"Sylvie Manguin",coverURL:"https://cdn.intechopen.com/books/images_new/3092.jpg",editedByType:"Edited by",editors:[{id:"50017",title:"Prof.",name:"Sylvie",surname:"Manguin",slug:"sylvie-manguin",fullName:"Sylvie Manguin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"72",title:"Ionic Liquids",subtitle:"Theory, Properties, New Approaches",isOpenForSubmission:!1,hash:"d94ffa3cfa10505e3b1d676d46fcd3f5",slug:"ionic-liquids-theory-properties-new-approaches",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/72.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"2270",title:"Fourier Transform",subtitle:"Materials Analysis",isOpenForSubmission:!1,hash:"5e094b066da527193e878e160b4772af",slug:"fourier-transform-materials-analysis",bookSignature:"Salih Mohammed Salih",coverURL:"https://cdn.intechopen.com/books/images_new/2270.jpg",editedByType:"Edited by",editors:[{id:"111691",title:"Dr.Ing.",name:"Salih",surname:"Salih",slug:"salih-salih",fullName:"Salih Salih"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"117",title:"Artificial Neural Networks",subtitle:"Methodological Advances and Biomedical Applications",isOpenForSubmission:!1,hash:null,slug:"artificial-neural-networks-methodological-advances-and-biomedical-applications",bookSignature:"Kenji Suzuki",coverURL:"https://cdn.intechopen.com/books/images_new/117.jpg",editedByType:"Edited by",editors:[{id:"3095",title:"Prof.",name:"Kenji",surname:"Suzuki",slug:"kenji-suzuki",fullName:"Kenji Suzuki"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3828",title:"Application of Nanotechnology in Drug Delivery",subtitle:null,isOpenForSubmission:!1,hash:"51a27e7adbfafcfedb6e9683f209cba4",slug:"application-of-nanotechnology-in-drug-delivery",bookSignature:"Ali Demir Sezer",coverURL:"https://cdn.intechopen.com/books/images_new/3828.jpg",editedByType:"Edited by",editors:[{id:"62389",title:"PhD.",name:"Ali Demir",surname:"Sezer",slug:"ali-demir-sezer",fullName:"Ali Demir Sezer"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"872",title:"Organic Pollutants Ten Years After the Stockholm Convention",subtitle:"Environmental and Analytical Update",isOpenForSubmission:!1,hash:"f01dc7077e1d23f3d8f5454985cafa0a",slug:"organic-pollutants-ten-years-after-the-stockholm-convention-environmental-and-analytical-update",bookSignature:"Tomasz Puzyn and Aleksandra Mostrag-Szlichtyng",coverURL:"https://cdn.intechopen.com/books/images_new/872.jpg",editedByType:"Edited by",editors:[{id:"84887",title:"Dr.",name:"Tomasz",surname:"Puzyn",slug:"tomasz-puzyn",fullName:"Tomasz Puzyn"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"6918",title:"New Ideas for in Vivo Detection of RNA",doi:"10.5772/7207",slug:"new-ideas-for-in-vivo-detection-of-rna",body:'\n\t\tThe parallel discovery of the catalytic potential of RNA by Tom Cech and Sydney Altman at the beginning of the 1980s completely changed our view of the capabilities of RNA molecules and won them the Nobel prize in 1989 (Kruger et al., 1982; Guerrier-Takada et al., 1983). Since then, many new roles have been discovered and assigned to RNA, including the ability to form complex multitasking, supramolecular machines, such as ribosomes or spliceosomes, in which RNAs play leading roles. While only about 1.5 % of the human genome codes directly for protein sequences, a large fraction of it is nonetheless transcribed to produce many noncoding RNAs (ncRNA) that carry out important cellular functions (International Human Genome Sequencing Consortium, 2004). The structures and functions of some ncRNA have been extensively characterized, for example, ribosomal RNAs (rRNA), transfer RNAs (tRNA), small nuclear and nucleolar RNAs (snRNA and snoRNA). Moreover, applications of recent technological advances are revealing new classes of ncRNA molecules with novel or yet unknown functions, ranging in size from very small (20-30 nts) to thousands of nucleotides long. Small ncRNA (~20-30nts in length) include microRNA (miRNA), Piwi-interacting RNA (piRNA), small interfering RNA (siRNA), trans-acting siRNA (ta-siRNA), natural antisense transcript siRNA (nat-siRNA) and small scan RNA (scnRNA) all of which help regulate various stages of gene expression (Choudhuri, 2009). The functions of “long noncoding RNA” are still not understood, but they seem to be involved in transcriptional and epigenetic gene regulation (Ponting et al., 2009).
\n\t\t\tMany ncRNAs form distinct yet very diverse three-dimensional structures to carry out their functions (Noller, 2005, Staple & Butcher, 2005, Montange & Batey, 2008). These functions include fundamental roles at all stages of gene expression including chromosome remodeling, transcriptional and translation regulation, mRNA processing, transport, and localization, protein translocation and posttranslational modification. RNA molecules use various mechanisms to participate in these processes. For example, miRNA, siRNA and piRNA inhibit gene expression by hybridizing to complementary sequences of target mRNAs in association with RISC ribonucleoprotein complexes and either temporarily block translation or target the mRNA for destruction (Carthew, 2006, Bushati & Cohen, 2007, Boyd, 2008). Riboswitches, usually located in the 5\' untranslated region (5’-UTR) of mRNA, also inhibit translation, but do so by forming specific three-dimensional structures, that respond to small molecule metabolites that are products of the pathway to which the gene belongs. Upon binding cognate metabolites, riboswitches undergo conformational changes in their secondary and tertiary structures which usually result in suppression of transcription or translation. Some riboswitches actually function as ribozymes, resulting in self-cleavage of the mRNA and complete inactivation of the mRNA (Montange & Batey, 2008, Winkler et al., 2004).
\n\t\t\tDefects in RNA expression or processing due to mutations and misregulation can lead to serious diseases (Cooper et al., 2009). Recent studies of miRNA suggest their important role in cancer biology (Lee & Dutta, 2009). The expression levels of specific miRNA are highly correlated with tumor, tissue type or disease stage. Aberrant miRNA levels are also involved in other diseases, including Tourette\'s syndrome, fragile X syndrome, myotonic dystrophy, and schizophrenia (Abelson, 2005, Caudy et al., 2002, Bilen et al., 2006; Perkins et al., 2007).
\n\t\t\tAll RNAs are produced by transcription in the nucleus, where most processing also takes place. Processing may require transport of RNA between specific locations in the nucleus (Vargas et al., 2005). Once mature, most RNAs are exported to the cytoplasm through the nuclear pores (Stewart, 2007, Kelly & Corbett, 2009), where some may be shuttled back to the nucleus (Hwang et al., 2007, Takano et al., 2005). Once in the cytoplasm, RNAs maybe further transported to specific sites to carry out their functions, depending on the cell type, its developmental stage, environmental signals or perturbations.
\n\t\tSeveral general methods for
Traditional methods for determining which RNAs are produced by a given cell population involve cell disruption and total RNA extraction and its analysis
Fluorescence-based techniques have gained popularity for several reasons. While radioactive labeling and autoradiography are potentially more sensitive, fluorescent-based methods have the advantages of 1) using labels with long shelf lives, 2) low toxicity and ease of handling, and 3) high resolution imaging at the single cell level. Moreover, a variety of fluorophores and quenchers, which differ in excitation and emission wavelength, fluorescence lifetime, and anisotropy, are commercially available. This makes fluorescent-based methods excellent candidates for high-throughput screening and real-time
Several recent reviews provide a critical evaluation of fluorescent methods that are currently available for live-cell RNA imaging (Bao et al., 2009; Rodriguez et al., 2007; Tyagi, 2009; Schifferer & Griesbeck, 2009, Raj & van Oudenaarden, 2009). Modern fluorescent visualization approaches can be subdivided into the two main groups with respect to how the fluorescent probe is delivered (Figure 1). The traditional and still most widely used method is fluorescent
Schematic representation of several strategies for
to the target increases specificity so long as (i) the melting temperature for exactly matching sequences is greater than physiological temperature and (ii) the melting temperature for single-base mismatches remains below this temperature. If the hybridization sequence is too short, unintended targets may also be bound thus decreasing specificity. If it is too long, one or more mismatches may be tolerated, decreasing specificity and reducing off-rates from correct targets and thus decreasing the biosensor’s responsiveness to decreases in levels of the target RNA.
\n\t\t\t\tTo increase
A number of innovations have been proposed to improve the signal-to-noise ratio of molecular beacons such as dual fluorescence resonance energy transfer (FRET) MBs and quenched autoligation (QUAL) probes (Santangelo et al., 2004; Santangelo et al., 2006; Satterfield et al., 2007). Dual FRET MBs involve a pair of quenched probes that bind to adjacent sites on the target RNA. One probe contains a FRET donor, and the other a FRET acceptor. When both are bound to the target, the donor can transfer its excitation energy to the acceptor, producing FRET emission. A further improvement to dual probes is the use of quenched autoligation (QUAL) probes which bind to adjacent sites and autoligate forming a single long probe that binds to the target with high specificity (Silverman and Kool, 2005). However QUAL probes have the disadvantage of binding essentially irreversibly to targets and hence cannot be used to monitor decreases in target RNAs dynamically.
\n\t\t\t\tQuantum dots, gold or silver nanoparticles and photoluminescent polymers were recently introduced as alternatives to organic fluorescent dyes for labeling hybridization probes (Algar et al., 2009; Kim et al., 2008). Quantum dots are brighter and have better resistance to photobleaching than organic fluorophores. Moreover, quantum dots possess broad absorption bands and narrow tunable emission bands that are desirable for optical multiplexing. In comparison with conventionally labeled probes, that have a tendency to accumulate rapidly in nuclei when microinjected, probes conjugated to quantum dots remain in the cytoplasm (Chen et al., 2007). Photoluminescent polymers and gold nanoparticles have better quenching efficiencies as compared to organic dyes (Kim et al., 2008).
\n\t\t\t\tDisadvantages of FISH and “quenched probe” techniques include the requirements for time-consuming and expensive chemical modifications of probe sequences to covalently attach fluorophore and quencher molecules. Moreover, these conjugates are not able to diffuse directly into cells and require perturbation or disruption of the membranes.
\n\t\t\tGFP and related autofluorescent proteins, proven immensely useful in visualizing proteins
To reduce the background signal from unbound RBP-GFP, the “protein fragment complementation” (PFC) or “split-GFP” methodology was introduced (Valencia-Burton et al., 2007). This method involves the application of rational protein design to separate the autofluorescent protein sequence into two parts that do not fluoresce individually and can be expressed separately. Each part is fused to a different RBP domain. In the presence of a target RNA that is engineered to contain adjacent binding motifs specific to the two RBPs, the two protein fragments are brought together to reconstitute an intact fluorescent protein. In addition to reduced background signal, this method has the virtue that it does not require introducing exogenous RNA fluorescent probes, and can therefore be considered a label-free method. The main disadvantage of this method is the fact that signal generation is essentially irreversible, limiting its use for real-time dynamics of RNA expression levels. An additional limitation of AFP-based methods is the relatively large size of the protein tag (~270 kDa for multiple copies), which raises concerns that transport and localization of the target RNA may be perturbed (Bao et al, 2009; Tyagi, 2009).
\n\t\t\t\t\tWe compare the features and advantages/disadvantages of the latter-mentioned methods for
Aptamers are functional, single-stranded RNA or DNA oligonucleotides, usually 30 to 100 nts in length, artificially selected from combinatorial libraries for high binding affinities to specific molecular targets. Combinatorial libraries of oligonucleotides, randomized at desired sequence positions and comprising up to 1016 different sequences, can be easily generated by modern nucleic acid synthesizers. The process of screening a randomized oligonucleotide pool to identify and amplify sequences that perform a specific function is called “Systematic Evolution of Ligands by Exponential Enrichment” (SELEX) and was introduced almost 20 years ago (Tuerk & Gold, 1990; Ellington & Szostak, 1990). SELEX has been used to obtain high affinity aptamers to almost any conceivable target and has even been adapted to obtain catalytic nucleic acids (ribozymes and DNAzymes) by selecting for
\n\t\t\t\tComparative analysis of in vivo RNA visualization methods. *-calculations based on 330 Da per nucleotide.
transition-state analogues (Weigand et al., 2006; Schlosser et al., 2006). This technology has therefore revolutionized the field of sensing chemistry by enabling researchers to systematically generate numerous aptamers targeting a diverse range of analytes. Large numbers of RNA and DNA aptamers have been obtained by this method in the past 20 years, targeting a broad range of small molecules including organic dyes, amino acids, cofactors, carbohydrates and nucleotides, as well as proteins and nucleic acids (Klussmann, 2006). Aptamers against different classes of antibiotics, proteins and even whole cells have already found wide applications in biosensing, diagnostics, drug development and nanomedicine, as recently reviewed by other authors (Liu et al., 2009; Cho et al., 2009).
\n\t\t\t\tAptamers provide a nucleic acid-based alternative technology to monoclonal antibodies for specific recognition of diverse molecular targets. Aptamer selection exhibits a number of advantages over monoclonal antibody technology including: 1) versatility in designing selection strategies; 2) lower cost; and 3) easy scale-up and production as no cell or animal culture is needed. Other advantages of using nucleic acid aptamers in biosensors include their relatively small sizes, high analyte specificities and high affinities (Kds down to pM range), low toxicities and immunogenicities, and amenability to chemical modification to suit the desired application. Finally, they are small enough that their 2D structures can be reliably predicted and their 3D structures solved by NMR solution methods, which facilitates “tuning up” their chemical and biological properties.
\n\t\t\t\tSELEX begins with chemical synthesis of a randomized DNA pool. If selection of an RNA aptamer is the goal, the DNA pool is transcribed prior to the selection procedure. Next, the pool is incubated with the target molecule to achieve efficient binding. This is followed by a carefully designed selection step to sequester those DNA or RNA oligonucleotides in the pool exhibiting the desired functional properties - this selection process being the most crucial step. Finally, the amplification step enriches the pool, and the entire cycle is repeated for a sufficient number of times to obtain a pool comprising only molecules with a desired functional property. The original SELEX procedure has been modified to achieve higher affinity and specificity of aptamer products (e.g. Negative SELEX, Counter SELEX), to improve selection towards more complex targets (e.g. Genomic SELEX, Deconvolution-SELEX, Tissue-SELEX) or to select aptamers with novel functional properties (e.g.Covalent SELEX, Photo-SELEX) (Stoltenburg et al., 2007).
\n\t\t\t\tThe Ellington group established a database of aptamers that is current as of 2006 and provides aptamer targets, sequences, details regarding selection procedures and references to original publications. It can be accessed at http://aptamer.icmb.utexas.edu/site (Lee et al., 2004).
\n\t\t\tOrganic dyes were the first targets used to develop the SELEX methodology for RNA and DNA (Ellington & Szostak, 1990; Ellington & Szostak., 1992). Recent work has been directed to obtain aptamers that target fluorogenic dyes, with the aim of developing new
This earlier work was subsequently adapted to obtain high-affinity aptamers that specifically bind to fluorogenic dyes, defined as molecules which exhibit minimal emission when free in solution, but which fluoresce intensely when tightly bound. Any strategy where an increase in emission signal occurs upon specific binding to a target is called a "light-up" methodology, and fluorogen-aptamer complexes are often referred to as “fluoromodules” or “light-up pairs”. The performance of sensing systems is usually quantified in terms of “fluorescence enhancement” and so it is important to define what is meant by this term. For classic molecular beacons or FRET probes, fluorescence enhancement is defined as the ratio of the emission of the probe bound to its cognate target divided by the emission of free probe in the absence of the target. For light-up dye/aptamer pairs, fluorescent enhancement is defined as the ratio of the emission of the light-up dye bound to its aptamer divided by the emission of the free dye in solution. For biosensors employing light-up dye/aptamer pairs allosterically linked to target recognition domains, fluorescence enhancement is defined as the ratio of the emission of the dye in the presence of both biosensor and target divided by the emission of the dye in the presence of the biosensor alone. In genetic fusions of dye-binding aptamers with target RNAs, fluorescence enhancement is the ratio of the emission of the dye bound to the aptamer fused with the target RNA divided by the emission of the dye in the presence of the target lacking aptamer modules.
\n\t\t\t\tWe focus the rest of this section on efforts to design light-up pairs and to adapt them for use as biosensors to detect and image specific molecules
Sando and coworkers suggested a new route for generating specific light-up pairs starting with a conventional fluorescent DNA-staining dye that exhibits a number of favorable characteristics, including cell permeability, solubility, and stability in cellular environments (Sando et al., 2007; Sando et al., 2008). The key idea to this approach was demonstrated using Hoechst dye 33258, a widely used fluorescent stain for dsDNA molecules that fluoresces upon binding to the minor groove of dsDNA. To adapt this dye for more specific light-up applications, a variety of chemical modifications of the dye were employed to reduce or completely suppress its nonspecific affinity for dsDNA. The most effective modification of the dye for this purpose was the addition of two t-Butyl groups to its terminal phenol ring (The resulting “Hoechst 33258 derivative” is shown in Figure 2C). Next, SELEX was used to obtain a DNA aptamer that bound Hoechst 33258 derivative with Kd = 878 nM and enhanced fluorescence emission by 191-fold (Sando et al., 2007). Further work obtained an improved RNA aptamer showing higher affinity for the dye (Kd=35 nM) but somewhat reduced emission enhancement (56-fold).
\n\t\t\t\tA similar strategy was adapted by Constantin and coworkers to design a novel fluorogenic cyanine dye, Dimethyl Indole Red (DIR) (Constantin et al., 2008). Cyanine dyes such as thiazole orange (TO) and its derivatives are fluorogenic intercalation dyes that comprise two heterocycles separated by a methine group. Substitution in TO of a bulky dimethylindole heterocycle in place of the thiazole ring sterically hindered intercalation. Addition of an anionic propylsulfonate group further minimized nonspecific interactions with nucleic acids by increasing electrostatic repulsion (DIR is shown in Figure 2C). An RNA aptamer was obtained for DIR by SELEX that binds with Kd=87 nM and 60-fold emission enhancement. An advantage of the cyanine dyes is that they have relatively narrow emission bands, their emission wavelength maxima easily varied by chemical modification. As a group, their emission maxima cover much of the visible spectrum and extend into the near infrared.
\n\t\t\t\tAnother recently developed approach for creating light-up dyes is to start with a dye that emits when free in solution and to covalently attach electron donor groups that quench its emission by a photo-induced electron transfer (PET) process (Sparano & Koide, 2005; Sparano & Koide, 2007). Aptamers that bind exclusively to the quencher moieties are then selected. The idea is to obtain aptamers that can block electron transfer and thereby restore emission of the fluorophore moiety. The PET approach was demonstrated in principle using 2\',7\'-dichlorofluorescein (DCF) as a fluorophore and N-(p-methoxyphenyl)-piperazine (MPP) as a quencher (Figure 2D). An RNA aptamer for the MPP quencher moiety was obtained by SELEX and was indeed found to enhance DCF fluorescence in the DCF/MPP conjugate in a concentration-dependent manner. As the authors pointed out, however, this particular DCF-MPP/aptamer system is unsuitable for
A novel class of light-up probes for RNA detection was developed by Stojanovic\'s group by conjugating nonspecific fluorogenic intercalating dyes such as thiazole orange (TO) to small molecules such as GMP and AMP, for which RNA aptamers have previously been obtained (Pei et al., 2009). The GMP aptamer binds the GMP-TO conjugate with Kd=60 nM and a 500-fold fluorescence enhancement (Figure 3). Similar results were obtained with an AMP aptamer and AMP-TO conjugate. To adapt this approach for
Light-up dye/aptamer pairs and their properties. A. Malachite Green (MG) aptamer 3D structure with basepairs annotated with Leontis-Westhof nomenclature. Watson-Crick Edges are shown in circles, Hoogsteen Edges in squares, and the Sugar Edges in triangles (
be visualized, and the small-molecule dye conjugate would have to be delivered into the cell by an appropriate mechanism (for a review of cellular delivery systems, see Bao et al., 2009). The main drawback of this approach is competition for aptamer binding by endogenous molecules (for example, cellular GMP or AMP). Nevertheless, this approach has great potential, as the number of small molecules that can be used here is almost limitless.
\n\t\t\tStojanovic’s strategy for RNA detection. Thiazole Orange (TO), a fluorescent intercalator, is conjugated to a small molecule (blue hexagon). RNA aptamer (pink region) specific for its target binds the small molecule and forces TO binding to double-stranded region at the specific location.
Some of the disadvantages of other RNA imaging methods (described above in section 2.1 and 2.2) may potentially be overcome using label-free approaches based on fluorogen-binding aptamers. First, aptamers can be entirely composed of unmodified RNA, allowing genetic fusion to genes coding for cellular RNAs of interest (Figure 4B). This has the advantage of endogenous synthesis of the targeting sequence. Moreover, these aptamers, even when used in multiple copies, do not significantly increase the size of the cellular RNA, and only the imaging dye needs to be introduced exogenously, as most organic dyes are able to diffuse into cells. Finally, dyes with suitable light-up characteristics and low affinity for cellular targets can be expected to give low background signals.
\n\t\t\t\tHowever, selecting appropriate sites for aptamer insertion remains a critical issue. To obtain valid imaging results one must ensure that the insertion of the aptamer molecules does not disrupt functionally important target RNA secondary or tertiary structure or protein-binding sites. In cases where the organism of interest is difficult to genetically modify, these aptamers can be coupled to a specific target recognition domain and introduced exogenously using a variety of delivery methods (Figure 4A).
\n\t\t\t\tAptamers have been conjugated to other aptamers forming “allosteric aptamers” or to ribozymes forming “aptazymes” (Soukup, 2004; Silverman, 2003). Stojanovic and Kolpashchikov first reported the design of novel allosteric sensing systems produced by coupling the MG-binding RNA aptamer through a “communication module” to each of a series of specific aptamers for ATP, FMN (flavin monomucleotide), and theophylline. These “allosteric aptamers” are capable of transducting binding information between the two aptamers (Stojanovic & Kolpaschikov, 2004). They showed that binding of the target molecule to its respective aptamer domain caused a significant conformational change in the RNA sensor that was relayed through the communication module to the MG aptamer module so as to enable it to bind MG and produce a fluorescent signal.
\n\t\t\t\t\tWithin a year, Kolpashchikov adapted this concept to create an allosteric RNA aptamer probe system to sense specific nucleic acid targets (Kolpashchikov, 2005). The MG aptamer domain of the probe was divided into two fragments, each of which also contained half of the sequence complementary to the target DNA (see Figure 5). In the presence of the target, the complementary “binding arms” of the binary probe hybridize cooperatively to the target, forming a three-way junction structure and reconstituting the MG aptamer by stabilizing the helix containing it. The probe-target complex then binds MG and renders it
\n\t\t\t\t\tLabel-free approaches for in vivo RNA imaging. A. Allosteric coupling of fluorogen-binding aptamer to another target recognition element. B. Genetic fusion of the fluorogen-binding aptamer to target RNA sequence.
Binary MG-aptamer based sensor (
fluorescent, signaling the presence of the analyte. A 20-fold fluorescence enhancement was reported for this biosensor in the presence of 2 M concentration of the target, with discrimination factors greater than 20-fold for half of the sequences differing from the intended analyte by single nucleotide substitutions. Sando and coworkers tested the applicability of Kolpashchikov’s strategy to design a binary probe using their novel light-up Hoechst derivative/DNA aptamer light-up pair (see section 2.2 above) (Sando et al., 2007). The same design principle was used: the DNA aptamer for the Hoechst dye was split into two molecules, each of which contains half of the sequence complementary to the target. When the target DNA binds to the complementary regions of the binary probe, a three-way junction forms which restores the DNA aptamer structure and allows it to bind to the Hoechst dye. These binary DNA probes showed 70-fold fluorescence enhancement; correct analyte was discriminated from single-mismatches by a factor of 15.
\n\t\t\t\t\tA major hurdle in designing probes for specifically detecting or imaging biological RNAs in complex environmental or whole cell samples is the presence of secondary structure in the target RNA, which may compete with probe binding, thus reducing the binding affinity of the biosensor. Paranemic binding between two pre-formed nucleic acid structures was originally developed for DNA nanotechnology by Seeman and coworkers (Shen et al., 2004) and subsequently demonstrated in RNA (Afonin et al., 2008). Paranemic binding provides the means to design probes targeting internal loop structures in the RNA analyte in a programmable, sequence-specific manner that obviates the need to unfold the target to expose extended single-stranded regions. Paranemic binding can be applied for recognition of symmetrical internal loops ranging in size from 4 to 8 nucleotides (Afonin et al., 2008), as well as asymmetrical loops which have different numbers of bases in each strand (Novikova et al, unpublished observations). Afonin
The key principle of the sensing strategies described above involves the incorporation of a fluorogen-binding aptamer within the probe structure in such a way that fluorogen binding depends upon the specific recognition and binding to the target.
\n\t\t\t\tParanemic RNA biosensor for folded RNA molecules (
In the previous section, we described strategies for RNA detection and visualization using hybridization probes allosterically coupled to fluorogen-binding aptamers. A related approach is to genetically encode the fluorogen- or fluorophore-binding aptamer directly into the gene sequence of a cellular RNA of interest. Sando
Eydeler
Zhang and co-workers inserted the MG aptamer sequence into the cloned gene of
Applications of light-up dye/aptamer pairs are not solely limited to sensing nucleic acids. Other applications for these technologies have been proposed and are under development. For example, MG dye was conjugated to kinesin-driven microtubules to develop mobile bioprobes that can capture MG aptamer-tagged mRNAs from a transcription mixture and sequester them in a microfluidic device for high-throughput screening (Hirabayashi et al., 2006).
\n\t\t\t\tTo date, remarkable progress has been made to visualize specific RNA molecules with high spatial and temporal resolution in living cells. These studies have begun to reveal detailed mechanisms of RNA trafficking, localization and turnover (Bratu et al., 2003, Lange et al., 2008; Zimyanin et al., 2008). However, significant challenges remain to render these methods more generally applicable and to overcome shortcomings of currently used methods. The ideal RNA biosensing system should be easy to deliver to living cells, non-toxic and non-perturbing to cellular functions. It should penetrate all parts of the cell homogeneously and be chemically stable in different cellular environments. The fluorescent reporter of the biosensor should have high absorptivity and quantum yield (brightness) as well as good photostability and low phototoxicity. The sensor system should be easy to implement, so that different RNA targets can be studied without extensive optimization. It should allow optical multiplexing so that interactions between two or more cellular RNAs (or RNA and protein) can be monitored. It should only fluoresce when bound to the intended target and otherwise produce low background fluorescence. It should bind specifically to the intended target with minimal binding to targets having similar sequence and should exhibit fast binding kinetics to monitor rapid changes in RNA concentration. Finally, it should allow for studying specific RNAs in whole tissues or organs. Of course, no current method possesses all these desired characteristics.
\n\t\t\tCurrently, hybridization probes offer the greatest variety and versatility in choice of fluorescent reporters as they can be conjugated to semiconductor quantum dots, in addition to a wide variety of organic dyes. However, they still require exogenous delivery which can perturb cell functions. Hybridization methods suffer from signal sensitivity issues for two reasons. Unmodified targets generally bind only one hybridization probe and degraded probes release fluorescent reporters. To minimize degradation, hybridization probes are usually synthesized with resistant, modified backbones (see above). To increase sensitivity, multiple copies of the complementary binding sequence can be introduced in the target gene (van den Bogaard & Tyagi, 2009). Whether or not the target is genetically engineered, the use of hybridization probes raises several concerns related to choosing appropriate binding sites on the target RNAs. First the target sites should have relatively weak secondary structures to allow the probe to bind with high affinity with minimal competition. Second the selected binding sites should not participate in crucial functions, especially related to the processing, localization, or regulation of the target RNA. Hybridization probes targeting RNA sequences that bind other cellular factors can potentially interfere with these processes and produce spurious imaging results.
\n\t\t\tAutofluorescent protein (AFP) probes are synthesized intra-cellularly from gene fusions directly encoded in cell lines or from plasmids that can be introduced by transfection. These methods are less perturbing than those used to deliver hybridization probes. Moreover, AFP probes can be directed to desired cellular compartments with appropriate peptide targeting sequences. However, the use of AFP probes requires insertion of RNA binding protein (RBP) recognition sites into target RNA sequences, which also requires attention to target RNA sequence to avoid interfering with RNA function, but is potentially less sensitive to RNA secondary structure. One disadvantage of the general AFP strategy is that unbound probes are also fluorescent. This has been largely solved by the “split-AFP“ approach, but this method has the disadvantage that signal generation is irreversible upon reconstitution of the fluorescent protein. Contrary to common perceptions, autofluorescent proteins also have phototoxicity issues, as under irradiation, they produce reactive oxygen species that can damage cells (Remington et al., 2006).
\n\t\t\tNew methods, employing fluorogen-binding aptamers, hold promise of addressing many of the shortcomings of the two currenty used methods. First, aptamer-based methods are generally simpler and potentially more flexible and can be readily adapted to the particular application. Like AFP probes and unlike hybridization probes, the aptamers themselves can be produced endogenously. They can be either genetically encoded in the target RNA or produced from plasmids. Of course, they can also be introduced exogenously like hybridization probes if that is required. Like hybridization probes and unlike AFP probes, there is no need for co-expressing additional fusion constructs that require additional time to mature and that need to be balanced in their expression to target RNA expression levels– a significant simplification of the procedure. Additionally, because fluorogen-binding aptamers can be genetically integrated in their targets, signal generation is rapidly and completely eliminated upon RNA degradation, thus reducing background and false-positive rates providing further improvement over both AFP and hybridization methods. Finally, the aptamer size is quite insignificant compared to protein-based probes. However, it is not yet well established how many tandem aptamers are required to achieve adequate sensitivity for single-molecule detection
In the 1830s, the use of plant resources such as flax, hemp and others was widespread as their fibers were in high demand by the textile, the paper and sailing industries. These plants were grown over large areas for exploitation. However, with the progress of science and technology (loom, steam engine, development of cotton harvesting and processing technique and others), materials such as metals, ceramics, glass, polymers, stones and concrete were preferred to plant resources. In 1900, fiber plants experienced their lowest implantation in terms of surface area. Indeed, the rise of new materials has greatly contributed to the improvement of human living conditions through the construction of more robust and sophisticated habitats, the development of the automotive, railway, aeronautics, textile industries, etc. Subsequently, in a concern for economy, lightness and performance, the development of composite materials was born during the 1930s.
The industrial use of plant fibers began in the early twentieth century with the manufacturing of aircraft seats, fuel tanks or other electronic boxes in plant fibers reinforced polymer materials. The need for securing constructions or structures that are made up of these materials inevitably arose. From an engineering viewpoint, this is taken into consideration during the design, due to a good knowledge of the material characteristics. Plant fibers have specific properties that make them good candidate reinforcing materials for high-performance composites and other applications [1]. However, the mechanical properties of PFs vary considerably both within the same species and from one species to another. Humidity variation, for example, leads to shrinkage or swelling that changes mechanical properties [2]. Similarly, their thermal properties are by far very different from those of synthetic fibers.
Various studies also indicate that plant fibers exhibit, for example, a very complex anisotropic behavior [3, 4, 5]. This anisotropy must be accounted for if a reliable design is to be achieved. Close collaboration between scientific disciplines such as botany, chemistry, biochemistry, molecular structural biology, plant genetics, physics and mechanics allows each of them to make a constructive and complementary contribution. PFs must withstand stresses of all kinds when they are associated with their deriving plants. They are loaded when it comes to supporting the weight of the plant or when it comes to resisting the winds, storms and hurricanes so common in their environment. PFs are diverse, and can all be studied for their use as engineering materials, in order to take benefit of the particular advantages offered by each of them. Meanwhile, their mechanical, physical and chemical characterization can differ between members of the same species and from one species to another. They are most often in the form of bundles (technical fibers) comprising one to twenty elementary fibers. They have a complex hierarchical structure inducing anisotropy and, have great geometric and mechanical variability. Humidity variation, for example, leads to shrinkage or swelling and changes in mechanical properties.
The characterization of material generally involves so-called monotonous tests (tensile, compression, torsion, bending or a combination) according to the load’s direction (uniaxial or multiaxial), cyclic tests, hardness and resilience tests. Tensile test is undoubtedly the most common test applied to PFs [6, 7, 8, 9, 10, 11, 12] because it allows obtaining Young’s modulus, strength and elongation at break. Recent works show that PFs exhibit a delayed behavior over time and temperature [13, 14] highlighting their viscoelastic nature. A tensile test alone is therefore not sufficient to characterize these materials.
This chapter is structured in four sections. Following this introduction, Section 2 will give an overview of some essential applications, the supply chain and the techniques of separating fibers from their plant. In Section 3, we will describe the experimental characterization methods generally used to derive their structure morphology and their elastic, viscoelastic and thermomechanical properties. Section 4 is the conclusion.
The natural fiber derived from wood, sisal, hemp, coconut, cotton, kenaf, flax, jute, abaca, banana leaf fibers, bamboo, wheat straw or other fibrous material and the matrix can be a polymeric material. The key advantage of natural fibers and their composites over traditional materials is their biological and environmental durability as well as their superior biodegradability. Natural PFs are increasingly used in several fields of engineering applications because of their interesting properties [15]. Diverse abundance of natural fiber, shapes and forms is caused by their occurrence in different climatic zones, hence stimulating the interest and opportunities to conduct comprehensive studies for identifying new applications for the fibers in industry. Notably, they are gaining popularity due to their optimal use in reinforcement of bio-composite structures. These fibers are biodegradable, structurally sound and environmental friendly. However, a sound theoretical basis for modeling their structure and mechanical behavior has yet to be established. Thus, it will be a priority field of study that will challenge the scientists and researchers.
The emerging trends and opportunities for natural fibers are broadening due to desirable attributes such as biodegradability, eco-friendly, sustainability and energy efficiency. Sustainability supply chain of natural fibers is assessed and rated based on the following criteria: water usage, CO2 emissions, cost, availability and any other impacts [16]. Moreover, in the fashion industry, businesses tend to identify the impacts of fibers on brands that contribute to the most impressive reduction in their impact on environmental footprint. Some of the preferred fibers include Linen, Tencel, Bamboo, Recycled Polyester, Recycled Wool, Cork, Organic Cotton and Hemp.
Perhaps the most important factor is the understanding of the entirety of the supply chain of natural fibers and the stages that contribute to having the biggest impacts. Consequently, a map of biodiversity quantitative impact indicators that help the companies determine where to focus their efforts in supply chain management to alleviate natural fiber environmental footprint was developed.
Nowadays, only 23% of companies take into account their environmental footprint when choosing their suppliers and between 40 and 60% of a company’s environmental footprint actually comes from its supply chain. Hence, in developing the natural fiber supply strategy, it is critical to understand the role of supply chain management and the associated impacts of environmental footprint. Network analysis, optimization of transhipment costs and decision analysis on optimal solutions to minimize both the supply chain cost and environmental footprint are essential toolkits in the advancement and promotion of natural fibers industry.
Moreover, over the last two decades, the trends in production of plant fibers have been declining due to popularity of synthetic fibers as well as adverse drought conditions. The fiber production plants spread across all continents of the globe. Table 1 illustrates the trends of different sources of fibers, production capacities and where they are produced.
Fiber source | World production (103 tonnes) | Origin | Country |
---|---|---|---|
Abaca | 70 | Leaf | Malaysia, Uganda, Philippines, Bolivia, Brazil |
Bambou | 10,000 | Stem | Africa, India, Brazil |
Banana | 200 | Stem | Africa, India, Brazil |
Broom | Abundant | Stem | |
Coir | 100 | Fruit | India, Sri Lanka, Philippines, Malaysia, Brazil |
Cotton Lint | 18,500 | Stem | India, Europe, USA |
Elephant Grass | Abundant | Stem | India, Africa |
Flax | 810 | Stem | Europe |
Hemp | 215 | Stem | Yugoslavia, China |
Jute | 2500 | Stem | India, Egypt, Guyana, Jamaica, Ghana, Malawi, Sudan, Tanzania, Brazil |
Kenaf | 770 | Stem | Iraq, Tanzania, Jamaica |
Linseed | Abundant | Fruit | USA |
Nettles | Abundant | Stem | Europe |
Oil Palm Fruit | Abundant | Fruit | Malaysia, India, Brazil, Indonesia, Philippines |
Palmyrah | Abundant | Stem | India |
Ramie | 100 | Stem | Honduras, Mauritius |
Roselli | 250 | Stem | Borneo, Guyana, Malaysia, Sri Lanka, Togo, Indonesia, Tanzania |
Rice Husk | Abundant | Fruit/grain | India, Japan, Brazil, Others |
Rice Straw | Abundant | Stem | India, Japan, Brazil, Others |
Sisal | 380 | Leaf | East Africa, Bahamas, Antigua, Kenya, Tanzania, India, Brazil |
Sun Hemp | 70 | Stem | Nigeria, Guyana, Siera Leone, India |
Wheat Straw | Abundant | Stem | USA, Brazil, India, Canada |
Wood | 1,750,000 | Stem | All Countries |
Fiber sources, country and annual production of plant fibers.
In 2018, world production of all apparel and textile fibers reached 110 million tons, with natural fiber production estimated at 32 million metric tons. Natural fibers accounted for 29% of the total world fiber production capacity, with most of annual yield variation linked to dry weather conditions. Moreover, the decline in the amounts of natural fibers in total fiber production in the last decade is due to the exponential growth in polyester production, whose demands were triggered by the fast-fashion apparel industry.
Cellulosic fibers originated from plants and trees such as cotton, flax, hemp, jute, ramie, kapok, coir and bamboo are termed natural PFs. Such fibers are derived from various parts of plants including leaves, stems (bast fibers), fruits and seeds. Because all natural PFs are made up of mainly cellulose, they are categorized as ‘natural cellulosic fibres’, which may consist of one plant cell or an aggregate of cells bounded together by non-cellulose materials. Major commercially used PFs include: seed fibers (cotton, coir, kapok), bast fibers (flax, hemp, ramie, bamboo, banana), leaf fibers (sisal, kenaf, pineapple, abaca). To date, bast fibers are produced and utilized to manufacture a wide array of traditional and novel products including ropes, nets, carpets, mats, brushes, mattresses, paper and board materials. Generally, PFs are classified into two groups, namely soft fibers and hard fibers. Soft fibers are obtained through labour-intensive processes. It involves the following steps: selection of plant and harvesting the plant, partial drying, pounding with stone mallet, scraped with devices similar to comb to clean the fibers, wash the fibers, dry in the sun and finally comb the fibers. Subsequently, the fibers are ready to be spun or twisted into thread or cord. Soft fibers are often used to make ropes, string, nets, bags, and hammocks.
Hard fibers are processed through successive phases of cutting, drying, cleaning, and soaking before they can be woven. They are strong and naturally flexible fibers, thus suitable and utilized to make furniture, birdcages, toys, baskets, and mats.
Figure 1(a) and (b) shows the matured flax plants grown under a controlled greenhouse environment and a setup of bench-scale trouph for water retting of flax stems [17].
Greenhouse controlled experiments for flax plants [
Historically, most plant fibers were extracted manually, supplemented by natural retting. Evidently, this process is tedious, time-consuming and the extracted quality of fibers depends on the skill of the labourer. Nowadays, these fibers are extracted by chemical, mechanical or biological methods.
Akubueze et al. [18], reviewed the chemical techniques employed to extract fibers from natural plants, which include alkali, acid and other reagents. The typical mechanical extraction methods involve the use of stripping the plant stem (typically known as Bacnis and Leonit processes). The latest mechanical extraction methods utilize the decortication process, whereby the plant stems are crushed between two drum rollers to obtain the fibers after removing the pulp. The use of decorticators increase fiber production by 20–25 times compared with the manual process. With biological processes, both consortium of microorganisms and enzymes are utilized to efficiently extract fibers from plant stems.
Overall, the mechanical extraction is incapable to remove the natural binding material (pectin) from the interspaces of the fibers within fiber bundle, chemical extraction is capable to remove the pectin within the fiber bundle but causes significant environmental pollution, whereas the biological extraction method provides increased fiber yield, with minimum detrimental effects to the environment.
According to the Centre for Learning and Teaching in Art and Design (CLTAD), bast fibers, for example, are generally obtained from the phloem, an inner skin of a plant. These fibers support the cells of the phloem to provide strength to the stem. During processing, the fibers need to be separated from both the interior (xylem) and exterior (epidermis) which is the outermost layer of cells. The processes for separating these fibers from plant stalks are known as retting and decortication. Bast fiber bundles are typically several feet long, composed of overlapping cellulose fibers and a cohesive gum (or pectin), which strengthens the stem of the plant. The processes with which the bast fibers are separated significantly influence the quality of fibers as there are many stages involved. Kumar et al. [19], reported that the processing of sustainable fiber starts with fiber extraction and yarn production followed by bleaching, dyeing, softening, printing and drying.
Moreover, the process that separates the fibers into smaller bundles and elementary fibers is known as retting. Fiber retting is a key process and is an important criterion that most industries value because it determines the ultimate properties of the fibers produced. Traditional retting methods include dew and water retting. Dew retting depends on ambient weather conditions, typically takes several weeks and hence the quality of fibers produced varies considerably. Similarly, water retting has been a primary method for low-cost production of bast fibers. The process involves submerging bast straws into water and then the decomposition of the pectic is effected by the activity of anaerobic microorganisms. The quality of retting is assessed by the weight, degumming rate and the fiber properties. The faster rate of weight loss is preferred, the degumming rate is evaluated as the percentage change in pectin content of phloem regions in the raw plant to those in water-retted plant, whereas the desired fiber properties include color, linear density and tensile strength. Ruan et al. [20], reported that water retting improved both whiteness and fineness as well as the mechanical properties of fibers.
Although water retting is capable to produce good quality fibers, the inherent long duration of 7–14 days and associated odor has made it less attractive. The retting period can be reduced to 100 h by using warm water (35°C), but high water consumption and unpleasant odor limit its use to some developing countries. Retting is the process by which pectin gets dissolved or softened from the fiber bundles and separates the fibers from stems through microbial activity. As such, a group of Clostridium microorganism is commonly known to play a significant role in the process by hydrolysing the pectin as it produces pectinase enzyme. These enzymes initially attack the cambium layer and then the other thin-walled cells in the cortex. This phenomenon takes place in most plant bast fibers as they have similar long filament structures, except those from cotton fibers which are single plant cells. As an example, for the retting process conducted in a bench-scale trouph under no-flow process water conditions, there were distinct features on how the fibers separate from bundles. Figure 2(a) and (b) show the scanning electron microscopy of the unretted and retted fibers of flax.
A SEM shows the microstructure of flax fibers (a) before retting and (b) after the retting process [
Figure 3(a) shows that cellulosic fiber production accounted for 6% of the total in 2018, synthetic filament accounted for 45% and synthetic staple 20%. Similarly, Figure 3(b) depicts that cotton accounted for 81% of natural fiber production by weight in 2018, jute accounted for 7%, while coir and wool each accounted for 3%.
World total fiber production and natural fiber production [
The synthetic fibers are dominated by polyester, which accounts for nearly 90% of world filament production and 70% of world synthetic staple production. The remaining synthetic fibers are composed mostly of nylon, acrylic and polypropylene.
Perhaps a key factor is to consider the role and contribution of human capital and household social economics. Employment statistics in natural fiber industries is difficult to estimate because households do not engage in consistent annual production. In Ref. [23] it is estimated that about 60 million households worldwide are engaged in natural fiber production, and hence the total employment, reflecting both full-time year-round employment and part-time or seasonal employment, is around 300 million, which represents about 4% of the world’s population.
Natural fibers possess superior advantages over synthetic fibers including widespread availability, low cost, low density, moderate strength modulus to weight ratio, high acoustic damping, low manufacturing energy consumption, low carbon footprint and biodegradability. Consequently, there are emerging concerted research initiatives that explore and promote the understanding of the characteristics of natural fibers [15, 24].
As discussed in Section 2.3.1 above, dew and water retting are the most common processes for fiber retting. Plant fibers can also be extracted using chemical and enzymatic retting, which provide better control than dew and water retting. Unfortunately, chemical retting while effective in extraction of fibers, causes significant pollution challenges due to higher amount of chemicals utilized. For the chemical extraction methods, alkali and selected reagents have been employed. Alkali treatments promote the fibrillation, whereby the composite fiber bundle is degraded into smaller fibers. Sodium hydroxide (NaOH) is popularly used to reduce the fiber roughness, but also produces good quality fiber. Reagents such as sulfuric acid, hydrogen peroxide, protease and sodium citrate can also be used for chemical extraction [25].
Similarly, enzymatic retting is relatively expensive despite its shorter retting time, yet it produces acceptable fiber quality and is advantageous over other retting processes. In the enzymatic method, the selection of enzymes depends on the type of substrate, composition, size and lignin content. The most common enzymes utilized are cellulases and pectinases. Cellulase enzymes enhance the fiber smoothness by removing fibrils from the outer layer. As such, this results in reduction in the mechanical properties due to the damage caused in the fibers. Pectinases remove the inter-lamellar pectin, which is a natural adhesive compound between fibers.
The ultrastructure is about dimensions between the atomic and molecular domains. These are accessed using microscopes. Morphology and quantitative chemistry investigations on plant fibers can be achieved following various analytical techniques such as Fourier transform infrared spectroscopy (FTIR), high-performance liquid chromatography (HPLC) and thermogravimetric analysis (TGA), surface electron microscopy (SEM), atomic force microscopy (AFM) and transmission electron microscopy (TEM) [7, 24]. TEM, which uses the principle of electron diffraction leads to very high magnifications of about 5,000,000. Recent progress in instrumentation has made Raman microscopy an extraordinary analytical tool in biological and plant research [26]. The main advantage of confocal Raman microscopy (CRM) is its lateral spatial resolution and the fact that it provides not only chemical composition information but also structural information.
A plant fiber is a nanostructured, renewable, sustainable and biodegradable composite material (Figure 4) [27]. Its cell wall can be likened to a composite lamina, consisting of a few plies reinforced with fibrils. Each individual fiber is composed of a primary wall P and a secondary wall S, itself consisting of three layers S1, S2, S3. In the centre, there may be a cavity called lumen if the cell has not filled up completely during its development. Individual cells are interphased with the middle lamellae (ML) as presented in Figure 4. The S2 layer of the secondary wall represents about 80% of the section and governs the mechanical behavior of the fiber [28]. The middle lamella is a wall 0.5–2 μm thick that surrounds the fiber; it plays the role of matrix that maintains the cohesion of the fibers. It is mainly composed of hemicelluloses, pectin and lignin (about 70%) [29]. Figure 5(a)–(d) show micrographs of the RC fiber [30] obtained on a Hitachi H-7650 TEM.
Simplified structure of the wood cell wall as seen by Coté [
TEM micrographs of the RC fiber (a) consecutive layers (16,400), (b) layer stacking (16,400), (c) warty sub-layer (7660) and (d) reinforcement by a small cell (10,900) [
The microfibrillar angle is defined as the angle that the microfibrils form with the longitudinal axis of the cell. These two parameters explain partially the difference in mechanical properties between different types of cortical fibers (Table 2) [15]. The microfibrillar angle has a major influence on the elastic properties of plant fibers. The weaker is this angle, the better are the properties for plant fibers to behave as a composite material, which presents better mechanical properties in the reinforcement direction [24, 31]. Xu and Liu [32] predicted that the cell wall elastic modulus of wood varies by a factor of 3 when microfibril angle changes from 40° to 10°.
Fibers | Crystallinity index (CrI) | Microfibril angle (°) | Cross-section area (mm2) | Length of the cell (mm) | Aspect ratio l/d |
---|---|---|---|---|---|
Coco | 45.0 | 1.20 | 3.3 | 35 | |
Flax | 10.0 | 0.12 | 2. | 1687 | |
Hemp | 6.2 | 0.06 | 23 | 960 | |
Jute | 8.0 | 0.12 | 2.3 | 110 | |
Ramie | 7.5 | 0.03 | 154 | 3500 | |
RC [35] | 42 | 0.05–0.962 | >2000 | ||
Sisal | 56.6–66.2 | 20.0 | 1.10 | 2.2 | 100 |
The cellulose fibrils are oriented in a helix at an angle called micro-fibril angle, as shown in Figure 4. The microfibril angle in the S1 and S3 layers is greater than that of the S2 layer. It means that the fibrils in S1 and S3 layers are almost transversely oriented with respect to the fiber axis. According to the small microfibril angle in the S2 layer, its fibrils are oriented more parallel to the axis of the fiber [22]. In addition, for a given percentage of cellulose, the lower the microfibril angle, the higher the stiffness and strength of the fiber. The greater the microfibril angle, the greater the elongation at break [28]. Each microfibril can be considered as chains of cellulose crystals bound by amorphous zones [36].
The microfibril angle partly explains the elastic deformation of the plant fiber and therefore its elongation at break. Under relatively low tensile forces, a plant fiber undergoes a reversible deformation due to the progressive alignment of cellulose microfibrils with the fiber axis and an elasto-visco-plastic deformation of amorphous polymers. If the stress of the fiber is stronger, the deformation of it enters an irreversible phase that can continue until the rupture. A high microfibril angle implies a greater elastic deformation for a low tensile fiber stress. In addition, there is a negative correlation between the microfibril angle and the corresponding Young’s modulus (Figure 6) [37].
Variation of the young modulus with the microfibril angle of a unit cell.
In order to estimate suitability of different fibers to engineering and other applications, it is necessary, among other things, to determine their mechanical properties in the longitudinal and transverse directions as well as the origin of the viscoelastic properties. Thus, we will present in the following paragraphs a state of the art on the main methods used to evaluate the elastic and viscoelastic properties of PFs. Various methods have been used to measure the angle of microfibrils in the S2 layer, which is generally considered a Z-helix. Nevertheless, some studies using cross-field pit punctuations such as those of Pysznski and Hejnowicz [38] on the tracheids of Norwegian Spruce show that in about 80% of the trees studied, the Z-shaped microfibrils have an angle of 10°–40° while in the remaining 20%, the angle is lower with variations in orientation. A complete list of the different microfibril angle measurement techniques with their advantages and disadvantages is given by Huang et al. [39]. Among these techniques, X-ray diffraction is fast, but it is impossible to measure the angle of a single fiber, because of the bundle, only an average of the angle on the X-rays affected cells can be determined. The results obtained by different methods are often contradictory. For example, the work of Herman et al. [40] on individual tracheids shows large variations in the microfibril angle within annual dark circles with a sharp decrease from spring cells to summer cells. While other studies by Lichtenegger et al. [41] using the SAXS (small-angle X-ray scattering) method, on the same cell type shows a higher microfibril angle in summer tracheids than in spring tracheids. Currently, it is necessary to understand where the differences in results obtained by the available measurement methods originate from and to find a method that gives safe and reproductive results. A technique was developed by Jang [42] which uses polarization confocal microscopy based on dichromic cell wall fluorescence when stained with specific fluorochromes showing a high affinity with cellulose. In this technique, sample preparation still needs to be addressed. In fact, very thin samples, only allow observation of fluorescence intensity in the S2 layer without interference with the other layers. A quick but reliable estimate of the Rhectophyllum Camerunense (RC) fiber [30] microfibrils angle was obtained on the SEM (following a microtome longitudinal section of the fiber coinciding with the S2 layer) and fluorescence micrographs.
The chemical composition of plant fibers depends largely on the particular needs of their stemming plant. However, cellulose, hemicellulose and lignin are the main constituents, and their content depends on the age, origin and extraction conditions of the fibers. Cellulose is the chemical constituent that contributes the most to the strength and stability of the plant cell wall and therefore of the fibers. The cellulose content of the fiber largely influences mechanical properties, the economic aspect and the production of the fiber. Fibers with a high cellulose content would be preferable for use in textiles, paper, composites and other fields of activity while those with a high hemicellulose content would be suitable for the production of ethanol and other fermentation products because hemicellulose is easy to hydrolyse in fermentable sugars. Thus, the value of plant fiber and its potential applications depends largely on its cellulose content. Let us say, however, that the value of a plant depends mainly on the quality of its fibers and their end-use and not on the cellulose content itself. As with all-natural products, mechanical and physical properties of natural fibers vary greatly. These properties depend on the chemical and structural composition which depends on the origin of extraction (from leaves, seeds or stems), the local environment where the plants grow, the age of the plants and the climate. The chemical composition, structure, defects and dimensions of the fiber cells are the main parameters that condition all properties of the fibers including mechanical properties [12]. With the exception of cotton, the constituents of plant fibers are cellulose, hemicellulose, lignin, pectin, waxes and water-soluble substances. The average chemical composition of some plant fibers is shown in Table 3.
Fiber | Chemical content (%) | |||
---|---|---|---|---|
Cellulose | Hemicellulose | Lignin | Pectin | |
Abaca | 63.2 | 19.6 | 5.1 | — |
Bamboo | 48 | 23 | 19 | — |
Cotton | 83 | 5 | — | — |
Flax | 65–70 | 10–16 | 2.9 | 2–4 |
Hemp | 67 | 16.1 | 4 | — |
Jute | 55–64 | 12–18 | 12–33 | 0.2 |
Kenaf | 55–59 | 18–20 | 6.8–8 | 4.5–5 |
Ramie | 68.6 | 13.1 | 0.6 | — |
RC | 68.2 | 16 | 15.6 | — |
Sisal | 54–66 | 12 | 7.3 | 0.8 |
TJ | 62.7 | 14.5 | 4.1 | 7.6 |
Wood | 83 | 5 | 19–26 | 0 |
Chemical contents of some fibers.
The chemical bonds of the fibers can be determined with FTIR. Crystallographic properties can be analyzed with XRD. TGA, DTA and DSC are used to understand the thermal degradation behavior, the maximum degradation temperature of fibers. Pull-out tests applied to both raw and NaOH treated fibers aim for evaluation of the surface interaction of fibers with polymer matrices for composite materials applications.
In 1838, Anselm Payen proposed that cell walls of many plant cells be made of the same substance to which he gave the name cellulose. Cellulose is a natural polymer whose molecule, formed by long chains, consists of units of D-anhydroglucopyranoses (formula: (C6 H10 O5)n) linked by β-(1,4)-glycosidic bonds in position C1 and C4 (Figure 7). It represents the most abundant biological molecule on our planet. It is present in plants, algae, bacteria and some animals.
Cellulose molecule.
Cellulose is the major constituent of wood and is the major constituent of cotton and other textile fibers such as flax, hemp, jute and ramie. Its degree of polymerization varies according to the plant species. It can be 14,000 for native cellulose, but the insulation and purification procedures reduce it very sharply by about 2500. Cellulose contributes to the strength and rigidity of the fiber thanks to its strongly oriented chains. These macromolecular chains can be arranged, either regularly, in crystalline regions, or randomly in amorphous regions. Mechanical properties of natural fibers depend on their type of cellulose, as each type has its own cellular geometry. If cellulose is a prime structural constituent for the vast majority of plant cell walls, then hemicellulose with lignin acts as binding materials. Properties depend on the fiber cell geometry of each type of cellulose and its degree of polymerization.
Hemicelluloses represent the second most abundant constituent of plant fiber. Hemicelluloses are polysaccharides found in lignocelluloses alongside cellulose and pectin. Hemicelluloses, unlike cellulose, are composed of several sugars that form short chains with ramifications. The sugars present can be divided into different groups: pentoses (xylose, arabinose), hexoses (glucose, mannose, galactose), hexo-uronic acids (glucuronic acid and methyl-glucuronic acid) and l-deoxyhexoses (rhamnose and fucose). Hemicelluloses are, by definition, water-soluble polysaccharides that can be extracted from the plant cell walls using alkaline solutions. They are the most hydrophilic biopolymers in the cell wall that promote moisture absorption. In their natural state, they have a degree of polymerization that varies from 200 to 300, and their structure depends on the plant species. The best-studied class of hemicelluloses are xyloglucans. They have a bridging role between cellulose microfibrils in order to strengthen the cell wall by interaction with cellulose and, in some cell walls, with lignin. They consist of a glucose chain and short side chains of xylose, galactose and fructose.
Lignin together with cellulose and hemicelluloses is part of the wood industry. Its proportion in wood varies between 15 and 30% [43]. Lignin or ‘lignins’ are three-dimensional polymers from the radical polymerization of three phenylpropenoic alcohols: coniferryl alcohol, sinapyl alcohol and p-coumaryl alcohol [44]. Lignin contributes to the rigidity of cell walls, and thus to the erect port of terrestrial higher plants. Lignin also offers a protective barrier against the microbial attack of plants. Indeed, due to its chemical nature, lignin is very resistant to various chemical agents and biological degradation. To sum up, lignin polymers make the cell wall rigid and impermeable, allowing the transport of water and nutrients through the vascular system by protecting plants from microbial invasion. Lignin is totally amorphous and hydrophobic. It is not hydrolysed by acids, but hot soluble in soda, easily oxidized and also condensable with phenol.
Pectins are polymers of acidic polysaccharides, composed of a main chain of uronic acid bound in 1–4. Regularly, rhamnose molecules are interspersed between these monomers by bonds 1–2 and 1–4. Some of these rhamnose units carry side chains composed of neutral oses among which galactose and arabinose are the most abundant. The type of bond between uronic acid and rhamnose molecules forms elbows. The pectin macromolecule appears like a zigzag. This arrangement contributes to its special properties and provides some flexibility to plants. Pectins are extracted from the fiber by a chemical method either by boiling water or by ethylene diamine tetraacetic acid.
Different methods can be used including solid pycnometers or gas pycnometers [45, 46, 47]. The choice of gases (helium for example) or immersion liquids such as toluene, ethanol and xylene is decisive for quality results [46, 47]. Fibers must be dried for at least 72 h in a desiccator containing silica (previously regenerated). Fibers are then cut into lengths of 5–15 mm and then introduced into the pycnometer which is eventually placed in the desiccator for at least 24 h. Before carrying out the hydrostatic weighing with the immersion liquids, the vortex agitation of fibers to evacuate the microbubbles between needs to be done. Significant degassing could occur at this stage and provides information on the porosity rate of the fibers [30].
In general, PFs are suitable for reinforcing plastics (thermosets and thermoplastics) and textiles manufacturing thanks to their relatively high strength and low density. The tensile strength and the modulus of elasticity of PFs are very important characteristics for the use of fibers as reinforcements in composite and textile materials. However, the tensile test data for most fibers in service have yet to be studied, as the data found in the literature are scattered and often unreliable. In fact, methods used for the characterization are not identical. Table 4 shows the tensile mechanical properties of some plant fibers compared to synthetic fibers [48]. The properties of the fibers and their structure depend on several factors such as the origin, variety, conditions of growth and harvesting of fibers associated with the treatments, the location in the stem, the presence or absence of a lumen, measurement techniques that vary greatly from one research team to another. These factors can make a difference for the same type of fiber and influence test results.
Fiber | Density (g/cm3) | Diametre (μm) | Length (mm) | Tensile strength (MPa) | Young modulus (GPa) | Elongation at break (%) | Moisture content (%) |
---|---|---|---|---|---|---|---|
Abaca | 1.5 | 10–30 (20) | 4.6–5.2 (4.9) | 430–813 (621.5) | 31.1–33.6 (32.35) | 2.9 | 14 |
Bamboo | 0.6–1.1 (0.85) | 25–88 (56.5) | 1.5–4 (2.75) | 270–862 (566) | 17–89 (53) | 1.3–8 (4.65) | 11–17 (14) |
Banana | 1.35 | 12–30 (21) | 0.4–0.9 (0.65) | 529–914 (721.5) | 27–32 (29.5) | 5–6 (5.5) | 10–11 (10.5) |
Coir | 1.2 | 7–30 (18.5) | 0.3–3 (1.65) | 175 | 6 | 15–25 (20) | 10 |
Cotton | 1.21 | 12–35 (23.5) | 15–56 (35.5) | 287–597 (442) | 6–10 (8) | 2–10 (6) | 33–34 (33.5) |
Flax | 1.38 | 5–38 (21.5) | 10–65 (37.5) | 343–1035 (689) | 50–70 (60) | 1.2–3 (2.1) | 7 |
Hemp | 1.47 | 10–51 (30.5) | 5–55 (30) | 580–1110 (845) | 30–60 (45) | 1.6–4.5 (3.05) | 8 |
Jute | 1.23 | 5–25 (15) | 0.8–6 (3.4) | 187–773 (480) | 20–55 (37.5) | 1.5–3.1 (2.3) | 12 |
Kenaf | 1.2 | 12–36 (24) | 1.4–11 (6.2) | 295–930 (612.5) | 22–60 (41) | 2.7–6.9 (4.8) | ) 6.2–12 (9.1) |
Pineapple | 1.5 | 8–41 (24.5) | 3–8 (5.5) | 170–1627 (898.5) | 60–82 (71) | 1–3 (2) | 14 |
Ramie | 1.44 | 18–80 (49) | 40–250 (145) | 400–938 (669) | 61.4–128 (94.7) | 2–4 (3) | 12–17 (14.5) |
RC | 0.94 | 70–350 (120) | — | 450–1500 (557.1) | 5.8 (±3.5) | 27.5 | — |
Sisal | 1.2 | 7–47 (27) | 0.8–8 (4.4) | 507–855 (681) | 9–22 (15.5) | 1.9–3 (2.45) | 11 |
TJ | (1.398) | 40–90 () | (404.0) | (32.3) | (1.8) |
Mechanical properties of some selected plant fibers versus synthetic fibers [48].
Selection of plant fiber implies a prior study of its mechanical properties, chemical resistance, dimensional stability, separation process, etc. It is worth recalling that linear cellulosic macromolecules are linked by hydrogen bonds and are closely associated with hemicelluloses and lignin, which confer stiffness to fiber. One of the issues of natural fibers is the scattered information and the differences in mechanical properties reported. Likewise, the lack of standards for both producers and users of these materials regarding methods to collect, process, post-process and characterize plant fibers underlines the complexity in the selection.
Quasi-static tensile test is the method commonly used in the literature for the characterization of the mechanical properties of plant fibers in the longitudinal direction. This type of characterization presents challenges linked to the assembly and to the single nature of the fiber. In addition, the geometry of the plant fiber makes it often difficult to conduct the tests. Therefore, evaluation of the mean diameter along the fiber using a microscope is necessary for the performance of the test. The single fiber is mounted on a paper frame and a drop of glue is used to stick the fibers. The role of this paper frame is to facilitate the handling and alignment of the fiber on the jaws of the experimental device as shown in Figure 8 [35].
Tensile test and gripping tab specimens for plant fibers.
The large dispersion of the mechanical properties of the plant fibers observed (Figure 9) is mostly related to the test conditions. The research work by Ntenga et al. [14] focused on the choice of the stress speed and the gage length, in order to keep the deformation in the elastic domain and reduce this dispersion during the tests. The machine cross-head speed of 1 mm/min and the gage length of 10 mm were found to cause less dispersion of the mechanical properties in a tensile test.
Tensile stress/strain curves for the four cross-head speeds of gage length 10 mm [
Nanoindentation is a technique used to characterize the longitudinal and transverse mechanical properties of fibers at the cell wall scale. Commonly measured properties are Young’s modulus and material hardness. In the literature, nanoindentation tests have been carried out to access both transverse and longitudinal mechanical properties on wood fibers [34, 49] and recently on flax fibers [50]. According to Cisse [51], nanoindentation only gives access to local behavior of the fiber, and the identification of mechanical properties requires knowledge and use of a behavior model. The testing technique consists of applying a force to the indenter and taking the area of the indentation, in order to determine the Young’s modulus and the hardness of the material (Figure 10(a) and (b)).
(a) Nano indentation experimental device and (b) indentor impression Berkovich [
A typical set of nanoindentation tests results [53] is shown in Figure 11.
Transverse modulus of plant fibers obtained in nano indentation.
Differences in transverse and longitudinal modulus noted between the fibers can be explained not only by the differences in micro-fibrillary angles but also by the rate of cellulose that varies between fibers. Hemp and sisal in particular have a cellulose content of around 60%, while that of flax is over 75%; however, the mechanical properties of cellulose are much superior to those of lignin, hemicelluloses and pectins, other constituents of natural fibers [50].
A large amount of work exists in the field of vibration-based non-destructive testing (NDT) including an extensive survey of over 300 papers by Kong et al. [54]. Indeed, the vibration-based technique has been a very active area of research for many years, however, has always dealt with rigid bodies. As an extension of the use of this technique, the purpose of this section is to present the applicability of the low-frequency vibration-based technique towards estimation of dynamic Young’s modulus of natural fiber-based materials, initially having no bending stiffness. This technique enhances the applicability of non-contact acoustic non-destructive testing to the estimation of dynamic characteristics of thin materials, where the current standard method [55] is not applicable.
Let us consider a thin rectangular specimen having a length
Specimen configuration (i): undeformed, and vibrating at (ii): fundamental frequency, (iii): second frequency in flexural mode.
The specimen, considered as a membrane, initially has no bending stiffness. It is then slightly stretched in the y-direction, in order to make it possible to vibrate transversally (i.e. in the
In general, for a specimen having intrinsic elasticity, the equation of motion is expressed as follows:
where
where
The frequency equation with the fixed-fixed boundary condition shown in Figure 12 above was derived in Mfoumou et al. [56] to obtain the frequency of vibration
where
For a plant fiber-based material considered as a membrane; therefore, no account of intrinsic elasticity is taken so that Eq. (3) is simplified, and the normal frequencies equation is expressed as:
The Young’s modulus can therefore be determined using the flexural resonance method by monitoring normal modes of vibration. These modes for an oscillating system are special solutions where all the parts of the system are oscillating with the same frequency. At these modes, considering only bending modes in the length direction (
thus, enabling extraction of the constant
Both creep experiment and relaxation experiment are two techniques commonly used to characterize the delayed behavior of ‘conventional’ materials. A creep test consists of imposing an almost instantaneous stress load on the plant fiber and maintaining it constantly over time and then proceeding to a discharge. The resulting deformation under the action of the load is creep, and that under the action of discharge is recovery. In general, the creep responses can be broken down into three stages depending on the strain rate as shown in the following Figure 13. The first stage in which creep occurs at a decreasing rate is called primary creep; the second step, commonly called secondary creep, is carried out at a relatively constant speed; and the third stage, tertiary creep, occurs at an increasing rate and terminates with material fracture.
Creep/recovery test of an elementary hemp fiber under a constant environment [
The creep test was successfully carried out on an elementary hemp fiber and the results allowed it possible to highlight the viscoelastic nature of the plant fiber [51]. Figure 13 shows the creep test results obtained.
When a constant strain is applied to a material for a long period, cross-links or the primary bonds that form between molecules start breaking with time and spontaneously lose their bonding capability. High level of strain or long period is the main reason for intermolecular bond breakage, thus creating stress decay over time, called stress relaxation. The rate of bond breakage influences the rate of stress relaxation. Other factors control the rate of bond breakdown, such as stress on the bond, chemical interference, molecular chain mobility which allows molecular chains to move out from their position. The behavior of stress relaxation in plant fibers is also influenced by temperature, humidity, and strain levels. The stress relaxation tests are therefore mainly performed with different ranges of temperature, humidity and strain levels. The time taken to reach the end of relaxation is called relaxation time. From other studies, it is reported that at higher temperature relaxation time becomes shorter, while at lower temperature it becomes longer but the shape of relaxation does not change with temperature [57]; moreover, the variation of strain level affects the stress relaxation [58]. The literature also reports the sensitivity of this class of material to loading-directionality, and ductile and brittle phenomena [59].
During structural design, the properties of the material must be considered. Elastic Modulus is one of the most important material properties describing the stiffness of the material. When a force is applied to an object, modulus of elasticity or elastic modulus gives the mathematical description of the object’s tendency to be deformed elastically.
In orthotropic materials such as wood-based natural fibers, the strain quickly increases linearly with the stress, then exhibit a nonlinear behavior when the strain exceeds the proportional limits. When the stress relaxation tests are conducted for a very small deformation, the viscoelasticity of the material can be considered linear. During stress relaxation test, the material relieves stress over time as well as the elastic modulus of material
where,
A rectangular strip of specimen is placed between the clamps of the tensile test machine (see Figure 8), and it is slightly loaded within its elastic region. The specimen is tested in uniaxial stress-state at a strain rate of 1 mm/mm with 0.4% strain changes. The elongation is kept constant at 0.4% strain level (1 mm extension) for 5400 s and time, stress, and strain are recorded.
Experiments were carried out for paperboard (PPR) without crack and PPR with crack. Five specimens were tested for each case and each experiment continued for 5400 s (1.5 h) with 1 mm extension. The reason for taking 1 mm extension was to keep the deformation within the elastic region.
The stress relaxation of each specimen was monitored and analyzed at constant.
elongation. The load, stress and time data for constant strain were obtained from the experiments. From the testing of five specimens in each case, we have plotted stress versus time curves. The plotted stress relaxation of PPR without and with the presence of a side crack is presented in Figure 14.
Stress relaxation of paperboard with and without crack.
Figure 15 show the stress relaxation behavior of PPR at different strain levels (two different extension levels, 1 mm and 0.5 mm).
Stress relaxation of paperboard for 1 mm and 0.5 mm extension.
The data obtained from the stress relaxation experiments are decreasing type of data with function of time and this type of data can be fitted to the poly-exponential function of the following form:
where,
The parameters of a set of mechanical models can be calculated from experimental data. MATLAB, for example, can be used to extract the parameters from the data. To analyze the suitability of the mechanical model with the experimental stress relaxation, Maxwell Model, Two-unit Maxwell Model, Modified Two-unit Maxwell Model, Standard linear solid model are constructed and then compared with the experimental relaxation. Analytical description of these models is given in [62].
In Ref. [56] we have chosen Foss method to develop curve fitting for all models and then compared with the experimental relaxation. Whereas in Ref. [15] we used the Zapas-Phillips method. The best-fitted model with the experimental data was then selected to analysis all experimental data and mathematically stress relaxation equations were derived.
To predict the stress relaxation behavior of natural fibers, we derived the mathematical equations for PPR with and without presence of crack. These equations were derived by the Modified Two-unit Maxwell model which suits best with the experimental result. Though we carried out our experimental tests with five specimens for each kind of test and among them three specimen-data were taken into consideration, but here we will construct the stress relaxation equation for only one specimen for each case.
Below the comparison, diagrams between experimental relaxation data and the Modified Two-unit Maxwell are shown in Figures 16 and 17. The stress relaxation equation for each case is derived using Modified Two-unit Maxwell model.
Stress relaxation of paperboard—curve fitting.
Stress relaxation of paperboard with crack–curve fitting.
Suitability of materials inverse characterization, destructive or non-destructive, is widely investigated [52, 63, 64]. Furtado et al. [65] used an ultrasound shear wave viscoelastography method to determine the viscoelastic complex shear modulus of macroscopically homogeneous tissues. Ilczyszyn et al. [66] performed the mechanical characterization of flax fibers using an inverse optimization simplex method.
The aim here is to use macro-micro approaches to achieve an efficient estimation of the fiber properties. In fact, homogenization laws of the micromechanics of the elastic/viscoelastic behavior of composite materials provide relationships of the properties of these materials in terms of their constituents’ properties. For an orthotropic material, the knowledge of its off-axes elastic modules in a set of
For a tensile test in the
There are five independent properties to be determined
Analytical expressions of the five properties in terms of fiber and matrix phase properties and the volume fractions are given by:
with
Eqs. (8)–(13) are then solved for
There are evolving global challenges on the utilization of non-renewable resources in the manufacturing industry and increasingly stringent environmental legislation. Both consumers and regulatory agencies are thriving for products that reduce dependency on fossil fuels and thus, are more environmentally friendly. As such, this paves for an opportunity to embrace the use of natural fibers in products and composites leading to significant growth of biobased economy, which the present chapter intends to stimulate.
The field of study of plant fibers that can be industrially exploited remains open. In this chapter, a particular emphasis has been put on their production, in particular on the methods that are generally used to separate them from their originating plants. To date, the question of improving the quality of the extracted fiber has been satisfactorily answered, particularly as regards the possibility of combining several methods when necessary. Some other questions still require research. These include, among others, growing conditions for seed multiplication and fiber production, harvesting methods, optimisation of fiber separation, the molecular basis for improving fiber decortication and performance. The knowledge gained from this work could be used to design new varieties of fibers, tailored for specific industrial applications. Similarly, the recourse to proteomics [68, 69], to isolate genes involved in the biosynthesis of cell wall lignin and hemicellulose in tobacco. Variations in these constituents can affect the fiber quality and cellulose availability. This could then lead to a new orientation on molecular selection research as well as genetic modifications studies to improve the quality of plant fibers.
Morphology and surface behavior of plant fibers are studied using various techniques such as XRD, FTIR, SEM, AFM, TEM and thermogravimetric analysis that helps in understanding the nature of natural fibers.
In terms of the mechanical behavior of plant fibers, important milestones have been achieved to highlight the influence of the chemical composition and structural parameters of the plant wall on their tensile properties. The microstructure of plant fibers is very complex, precisely when it comes to defining generalizable geometric and analytical models that describe it. As mentioned above, improving the mechanical properties of fibers may require the introduction of new types of fibers. And we could mention in this regard the ongoing research on spinning with solvents [70, 71], to obtain fibers of greater strength and low scattered properties. Understanding how fiber morphology affects the properties of composite materials is essential. More precisely, it is important for the selection of new fibers and for the cultivation of fibrous plants genetically selected. This would help to predict their potential for reinforcement in other materials to achieve desired properties.
Investigation of the viscoelastic properties of plant fibers has also been outlined. A variety of dynamic modulus measurement methods exists including ultrasonic wave propagation and the flexural resonance method presented here, for which normal modes of vibration are monitored. Stress relaxation tests are to be carried out to retrieve stress over time as well as the elastic modulus of the fiber material. A mathematical method for extracting the relaxation modulus from relaxation experimental data has to be proposed to this end. Proper selection of the testing vibrational mode and machine cross-head speed (during relaxation) appear important in the suggested methods in order to avoid dispersive results. The Young’s modulus that is obtained from the dynamic behavior of the specimen should, therefore, reflects the frequency dependence of the material.
The authors wish to acknowledge the Director of the University Institute of Technology of the University Ngaoundéré, Prof. Mohammadou Bouba Adji, for providing research facilities within the department of mechanical engineering.
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Morrish"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7121",title:"Cell Growth",subtitle:null,isOpenForSubmission:!1,hash:"9845b4d66ecca197908bcbfe4fd89321",slug:"cell-growth",bookSignature:"Biba Vikas and Michael Fasullo",coverURL:"https://cdn.intechopen.com/books/images_new/7121.jpg",editedByType:"Edited by",editors:[{id:"241658",title:"Dr.",name:"Biba",middleName:null,surname:"Vikas",slug:"biba-vikas",fullName:"Biba Vikas"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8498",title:"Extracellular Vesicles and Their Importance in Human Health",subtitle:null,isOpenForSubmission:!1,hash:"eb168770441543e33da9325f16197fb4",slug:"extracellular-vesicles-and-their-importance-in-human-health",bookSignature:"Ana Gil De Bona and Jose Antonio Reales Calderon",coverURL:"https://cdn.intechopen.com/books/images_new/8498.jpg",editedByType:"Edited by",editors:[{id:"203919",title:"Dr.",name:"Ana",middleName:null,surname:"Gil De Bona",slug:"ana-gil-de-bona",fullName:"Ana Gil De Bona"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8774",title:"Programmed Cell Death",subtitle:null,isOpenForSubmission:!1,hash:"0459d0c7a518f61817a48fd4709c35bd",slug:"programmed-cell-death",bookSignature:"Hala Gali-Muhtasib and Omar Nasser Rahal",coverURL:"https://cdn.intechopen.com/books/images_new/8774.jpg",editedByType:"Edited by",editors:[{id:"57145",title:"Prof.",name:"Hala",middleName:null,surname:"Gali-Muhtasib",slug:"hala-gali-muhtasib",fullName:"Hala Gali-Muhtasib"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6883",title:"Cell Signalling",subtitle:"Thermodynamics and Molecular Control",isOpenForSubmission:!1,hash:"e4e17d85c0643c7f4d274fa9adbcc628",slug:"cell-signalling-thermodynamics-and-molecular-control",bookSignature:"Sajal Ray",coverURL:"https://cdn.intechopen.com/books/images_new/6883.jpg",editedByType:"Edited by",editors:[{id:"173697",title:"Prof.",name:"Sajal",middleName:null,surname:"Ray",slug:"sajal-ray",fullName:"Sajal Ray"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6925",title:"Endoplasmic Reticulum",subtitle:null,isOpenForSubmission:!1,hash:"a9e90d2dbdbc46128dfe7dac9f87c6b4",slug:"endoplasmic-reticulum",bookSignature:"Angel Català",coverURL:"https://cdn.intechopen.com/books/images_new/6925.jpg",editedByType:"Edited by",editors:[{id:"196544",title:"Prof.",name:"Angel",middleName:null,surname:"Catala",slug:"angel-catala",fullName:"Angel Catala"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6964",title:"Cell Culture",subtitle:null,isOpenForSubmission:!1,hash:"045f3a964a9628162956abc06ef5777d",slug:"cell-culture",bookSignature:"Radwa Ali Mehanna",coverURL:"https://cdn.intechopen.com/books/images_new/6964.jpg",editedByType:"Edited by",editors:[{id:"182118",title:"Dr.",name:"Radwa Ali",middleName:null,surname:"Mehanna",slug:"radwa-ali-mehanna",fullName:"Radwa Ali Mehanna"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6820",title:"Keratin",subtitle:null,isOpenForSubmission:!1,hash:"6def75cd4b6b5324a02b6dc0359896d0",slug:"keratin",bookSignature:"Miroslav Blumenberg",coverURL:"https://cdn.intechopen.com/books/images_new/6820.jpg",editedByType:"Edited by",editors:[{id:"31610",title:"Dr.",name:"Miroslav",middleName:null,surname:"Blumenberg",slug:"miroslav-blumenberg",fullName:"Miroslav Blumenberg"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7264",title:"Calcium and Signal Transduction",subtitle:null,isOpenForSubmission:!1,hash:"e373a3d1123dbd45fddf75d90e3e7c38",slug:"calcium-and-signal-transduction",bookSignature:"John N. Buchholz and Erik J. Behringer",coverURL:"https://cdn.intechopen.com/books/images_new/7264.jpg",editedByType:"Edited by",editors:[{id:"89438",title:"Dr.",name:"John N.",middleName:null,surname:"Buchholz",slug:"john-n.-buchholz",fullName:"John N. Buchholz"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],booksByTopicTotal:15,seriesByTopicCollection:[],seriesByTopicTotal:0,mostCitedChapters:[{id:"64565",doi:"10.5772/intechopen.81552",title:"Two-Dimensional (2D) and Three-Dimensional (3D) Cell Culturing in Drug Discovery",slug:"two-dimensional-2d-and-three-dimensional-3d-cell-culturing-in-drug-discovery",totalDownloads:3324,totalCrossrefCites:11,totalDimensionsCites:40,abstract:"Cell culture is an indispensable in vitro tool used to improve our perception and understanding of cell biology, the development of tissue engineering, tissue morphology, mechanisms of diseases and drug action. Efficient cell culturing techniques both in vitro and in vivo allow researchers to design and develop new drugs in preclinical studies. Two-dimensional (2D) cell cultures have been used since 1900s and are still a dominant method in many biological studies. However, 2D cell cultures poorly imitate the conditions in vivo. Recently three-dimensional (3D) cell cultures have received remarkable attention in studies such as drug discovery and development. Optimization of cell culture conditions is very critical in ensuring powerful experimental reproducibility, which may help to find new therapies for cancer and other diseases. In this chapter, we discuss the 2D and 3D cell culture technologies and their role in drug discovery.",book:{id:"6964",slug:"cell-culture",title:"Cell Culture",fullTitle:"Cell Culture"},signatures:"Jitcy Saji Joseph, Sibusiso Tebogo Malindisa and Monde Ntwasa",authors:null},{id:"68141",doi:"10.5772/intechopen.87778",title:"Nonenzymatic Exogenous and Endogenous Antioxidants",slug:"nonenzymatic-exogenous-and-endogenous-antioxidants",totalDownloads:1920,totalCrossrefCites:17,totalDimensionsCites:30,abstract:"Nonenzymatic exogenous and endogenous antioxidants play an important role in human health and act as preservatives for cosmetics, pharmaceuticals, and food products. This chapter will discuss the chemical structure and mechanism of action of the most important nonenzymatic small exogenous and endogenous organic molecules that act as antioxidants. The chapter will focus on the structural features, functional groups, properties, biosynthetic origin, and mechanism of action of such antioxidants. It also covers damages that free radicals create and the mechanisms by which they are neutralized by the various antioxidants. The scope of this chapter will be limited to nonenzymatic exogenous and endogenous antioxidants since enzymatic antioxidants have been discussed extensively in several reviews.",book:{id:"7999",slug:"free-radical-medicine-and-biology",title:"Free Radical Medicine and Biology",fullTitle:"Free Radical Medicine and Biology"},signatures:"Ziad Moussa, Zaher M.A. Judeh and Saleh A. Ahmed",authors:[{id:"300774",title:"Dr.",name:"Ziad",middleName:null,surname:"Moussa",slug:"ziad-moussa",fullName:"Ziad Moussa"},{id:"306324",title:"Dr.",name:"Zaher",middleName:null,surname:"M. A. Judeh",slug:"zaher-m.-a.-judeh",fullName:"Zaher M. A. Judeh"},{id:"306325",title:"Prof.",name:"Saleh",middleName:null,surname:"A. Ahmed",slug:"saleh-a.-ahmed",fullName:"Saleh A. Ahmed"}]},{id:"62562",doi:"10.5772/intechopen.79502",title:"Keratin Waste: The Biodegradable Polymers",slug:"keratin-waste-the-biodegradable-polymers",totalDownloads:2288,totalCrossrefCites:9,totalDimensionsCites:16,abstract:"Keratins are everywhere, from being the major components of household dust to common contaminants of laboratory protein analysis. Keratin is the major structural fibrous protein belonging to the large family of structural proteins to form hair, wool, feathers, nails, and horns of many kinds of animals and has a high concentration of cysteine, 7–20% of the total amino acid residues, that form inter- and intramolecular disulfide bonds. Keratin wastes are considered as the environmental pollutants and produced mostly from the poultry farms, slaughterhouses, and leather industries. Keratin wastes are dumped, buried, used for landfilling, or incinerated and all these actions increase the threats of environmental hazards, pollution, negatively influence the public health, and increase greenhouse gases concentration. Nature has provided planet Earth with a variety of beneficial organisms. Soil is considered as a well-known source for the growth of keratinophilic microflora (fungi and bacteria), which have the capability to degrade the keratin waste. The keratin-degradation ability of keratinophilic microflora has been credited with the production of the microbial keratinase enzyme and biodegradation takes place (enzymatic degradation). So, the keratin wastes are the biodegradable polymers. Keratinase is the industrially significant enzyme that offers bioconversion of keratin waste, utilization as animal feed supplements, and dehairing agents in tannery industries and textile industries.",book:{id:"6820",slug:"keratin",title:"Keratin",fullTitle:"Keratin"},signatures:"Tarun Kumar Kumawat, Anima Sharma, Vishnu Sharma and\nSubhash Chandra",authors:[{id:"250905",title:"Dr.",name:"Anima",middleName:null,surname:"Sharma",slug:"anima-sharma",fullName:"Anima Sharma"},{id:"257932",title:"Dr.",name:"Tarun Kumar",middleName:null,surname:"Kumawat",slug:"tarun-kumar-kumawat",fullName:"Tarun Kumar Kumawat"},{id:"257942",title:"Dr.",name:"Vishnu",middleName:null,surname:"Sharma",slug:"vishnu-sharma",fullName:"Vishnu Sharma"},{id:"257944",title:"Prof.",name:"Subhash",middleName:null,surname:"Chandra",slug:"subhash-chandra",fullName:"Subhash Chandra"}]},{id:"62159",doi:"10.5772/intechopen.79050",title:"Keratins in Skin Epidermal Development and Diseases",slug:"keratins-in-skin-epidermal-development-and-diseases",totalDownloads:2435,totalCrossrefCites:5,totalDimensionsCites:13,abstract:"Epidermal keratinocyte (KC), the major cell type in the skin epidermis, plays critical roles in forming a permeability barrier to separate internal organs from external stimuli. Keratins, constituting about 30–80% of the total protein in KCs, form the major intermediate filament cytoskeleton of KC. Keratins consist of 54 unique genes in humans and they are expressed in cell-, differentiation- and development-dependent manner. While keratin pairs K5-K14 and K1-K10 are normally associated with KCs at different cell differentiation stages, other keratin pairs such as K6-K16/K17 and K8–K18 and are usually not expressed in normal skin interfollicular epidermis, but are elevated during wounding, inflammatory skin diseases such as psoriasis or malignant conversion of KC. The expression and function of keratins are tightly regulated at both transcriptional and post-transcriptional levels. Inherited or spontaneous mutations in keratins or abnormal keratin regulations or modifications can cause KC and cutaneous tissue fragility, skin hypertrophic and inflammatory conditions or malignant transformation of KC, therefore accounting for a large number of disorders in human skin. Here we review the recent literature on how keratins are normally expressed during skin development and how mutations or misregulations of these keratins are involved in the pathogenesis of skin diseases.",book:{id:"6820",slug:"keratin",title:"Keratin",fullTitle:"Keratin"},signatures:"Ling-juan Zhang",authors:[{id:"241614",title:"Dr.",name:"Lingjuan",middleName:null,surname:"Zhang",slug:"lingjuan-zhang",fullName:"Lingjuan Zhang"}]},{id:"63131",doi:"10.5772/intechopen.80051",title:"Keratinaceous Wastes and Their Valorization through Keratinolytic Microorganisms",slug:"keratinaceous-wastes-and-their-valorization-through-keratinolytic-microorganisms",totalDownloads:1455,totalCrossrefCites:4,totalDimensionsCites:10,abstract:"Keratin is a fibrous protein mainly found in higher vertebrates such as mammals, birds, and reptiles. It is also a major constituent of human epithelial tissues. Major keratinaceous wastes include skin, hair, wool, feather, horns, hooves, and nails. Large amounts of such wastes are generated from meat industry, poultry houses, and wool industry etc. Though keratinous wastes contain about 90% protein, keratin is usually recalcitrant to normal proteases. Such wastes have been traditionally digested using physico-chemical methods. But such techniques are energy-intensive and technologically demanding. Also, such approaches lead to degradation of certain amino acids such as lysine. In nature, keratinaceous wastes don’t accumulate indicating that keratinolytic microorganisms exist in nature. Keratinase producing strains are distributed among bacteria, fungi, and actinobacteria etc. Hence, potent keratinolytic microbes and their enzymes may be used for valorization of keratinous wastes. Efficient degradation of such wastes may generate value-added products such as feed additives, agricultural biofertilizers, and cosmetics. This chapter will give a comprehensive overview of types of keratinaceous wastes, kinds of keratinolytic microbes and keratinases, and valorization of such wastes using keratinase producing strains and/or keratinases.",book:{id:"6820",slug:"keratin",title:"Keratin",fullTitle:"Keratin"},signatures:"Debananda Singh Ningthoujam, Keishing Tamreihao, Saikat\nMukherjee, Rakhi Khunjamayum, Laishram Jaya Devi and Roshan\nSingh Asem",authors:[{id:"224389",title:"Dr.",name:"K",middleName:null,surname:"Tamreihao",slug:"k-tamreihao",fullName:"K Tamreihao"},{id:"242319",title:"Prof.",name:"Debananda",middleName:null,surname:"Ningthoujam",slug:"debananda-ningthoujam",fullName:"Debananda Ningthoujam"},{id:"262045",title:"Dr.",name:"Saikat",middleName:null,surname:"Mukherjee",slug:"saikat-mukherjee",fullName:"Saikat Mukherjee"},{id:"262046",title:"Ms.",name:"Rakhi",middleName:null,surname:"Khunjamayum",slug:"rakhi-khunjamayum",fullName:"Rakhi Khunjamayum"},{id:"262076",title:"Ms.",name:"Laishram",middleName:null,surname:"Jaya Devi",slug:"laishram-jaya-devi",fullName:"Laishram Jaya Devi"},{id:"262077",title:"Mr.",name:"Roshan Singh",middleName:null,surname:"Asem",slug:"roshan-singh-asem",fullName:"Roshan Singh Asem"}]}],mostDownloadedChaptersLast30Days:[{id:"67793",title:"Kinetic Studies on Cell Growth",slug:"kinetic-studies-on-cell-growth",totalDownloads:3810,totalCrossrefCites:3,totalDimensionsCites:8,abstract:"The kinetic model of cell growth is substantially capable to predict product formation. Mathematical models provide a strategy for solving problems encountered in fermentation process. A biochemical engineering approach to address this problem could be to develop a mathematical model which not only helps in the understanding of the system but also predicts various cultivation strategies to facilitate the optimization of a fermentation process, saving much of the time and cost for performing experiments. The presented overview indicates that many of the environmentally relevant aspects in growth kinetics are still waiting to be discovered, established, and exploited. A kinetic model that describes microbial growth, product formation and substrate consumption and the experimental data were fitted with modified logistic equation.",book:{id:"7121",slug:"cell-growth",title:"Cell Growth",fullTitle:"Cell Growth"},signatures:"Punniavan Sakthiselvan, Setti Sudharsan Meenambiga and Ramasamy Madhumathi",authors:[{id:"268626",title:"Dr.",name:"Punniavan",middleName:null,surname:"Sakthiselvan",slug:"punniavan-sakthiselvan",fullName:"Punniavan Sakthiselvan"},{id:"269591",title:"Dr.",name:"Madhumathi",middleName:null,surname:"Ramasamy",slug:"madhumathi-ramasamy",fullName:"Madhumathi Ramasamy"},{id:"279920",title:"Dr.",name:"S S",middleName:null,surname:"Meenambiga",slug:"s-s-meenambiga",fullName:"S S Meenambiga"}]},{id:"69690",title:"Ion Homeostasis Response to Nutrient-Deficiency Stress in Plants",slug:"ion-homeostasis-response-to-nutrient-deficiency-stress-in-plants",totalDownloads:2552,totalCrossrefCites:3,totalDimensionsCites:6,abstract:"A crucial feature of plant performance is its strong dependence on the availability of essential mineral nutrients, affecting multiple vital functions. Indeed, mineral-nutrient deficiency is one of the major stress factors affecting plant growth and development. Thereby, nitrogen and potassium represent the most abundant mineral contributors, critical for plant survival. While studying plant responses to nutrient deficiency, one should keep in mind that mineral nutrients, along with their specific metabolic roles, are directly involved in maintaining cell ion homeostasis, which relies on a finely tuned equilibrium between cytosolic and vacuolar ion pools. Therefore, in this chapter we briefly summarize the role of the ion homeostasis system in cell responses to environmental deficiency of nitrate and potassium ions. Special attention is paid to the implementation of plant responses via NO3− and K+ root transport and regulation of ion distribution in cell compartments. These responses are strongly dependent on plant species, as well as severity and duration of nutrient deficiency.",book:{id:"7121",slug:"cell-growth",title:"Cell Growth",fullTitle:"Cell Growth"},signatures:"Natalia Osmolovskaya, Julia Shumilina, Ksenia Bureiko, Veronika Chantseva, Tatiana Bilova, Ludmila Kuchaeva, Nikolai Laman, Ludger A. Wessjohann and Andrej Frolov",authors:[{id:"177609",title:"Dr.",name:"Natalia",middleName:null,surname:"Osmolovskaya",slug:"natalia-osmolovskaya",fullName:"Natalia Osmolovskaya"},{id:"309520",title:"Ms.",name:"Julia",middleName:null,surname:"Shumilina",slug:"julia-shumilina",fullName:"Julia Shumilina"},{id:"309521",title:"Ms.",name:"Ksenia",middleName:null,surname:"Bureiko",slug:"ksenia-bureiko",fullName:"Ksenia Bureiko"},{id:"309522",title:"Ms.",name:"Veronika",middleName:null,surname:"Chantseva",slug:"veronika-chantseva",fullName:"Veronika Chantseva"},{id:"309523",title:"Dr.",name:"Tatiana",middleName:null,surname:"Bilova",slug:"tatiana-bilova",fullName:"Tatiana Bilova"},{id:"309524",title:"Mrs.",name:"Ludmila",middleName:null,surname:"Kuchaeva",slug:"ludmila-kuchaeva",fullName:"Ludmila Kuchaeva"},{id:"309525",title:"Prof.",name:"Ludger A.",middleName:null,surname:"Wessjohann",slug:"ludger-a.-wessjohann",fullName:"Ludger A. Wessjohann"},{id:"309526",title:"Dr.",name:"Andrej",middleName:null,surname:"Frolov",slug:"andrej-frolov",fullName:"Andrej Frolov"}]},{id:"62159",title:"Keratins in Skin Epidermal Development and Diseases",slug:"keratins-in-skin-epidermal-development-and-diseases",totalDownloads:2435,totalCrossrefCites:5,totalDimensionsCites:13,abstract:"Epidermal keratinocyte (KC), the major cell type in the skin epidermis, plays critical roles in forming a permeability barrier to separate internal organs from external stimuli. Keratins, constituting about 30–80% of the total protein in KCs, form the major intermediate filament cytoskeleton of KC. Keratins consist of 54 unique genes in humans and they are expressed in cell-, differentiation- and development-dependent manner. While keratin pairs K5-K14 and K1-K10 are normally associated with KCs at different cell differentiation stages, other keratin pairs such as K6-K16/K17 and K8–K18 and are usually not expressed in normal skin interfollicular epidermis, but are elevated during wounding, inflammatory skin diseases such as psoriasis or malignant conversion of KC. The expression and function of keratins are tightly regulated at both transcriptional and post-transcriptional levels. Inherited or spontaneous mutations in keratins or abnormal keratin regulations or modifications can cause KC and cutaneous tissue fragility, skin hypertrophic and inflammatory conditions or malignant transformation of KC, therefore accounting for a large number of disorders in human skin. Here we review the recent literature on how keratins are normally expressed during skin development and how mutations or misregulations of these keratins are involved in the pathogenesis of skin diseases.",book:{id:"6820",slug:"keratin",title:"Keratin",fullTitle:"Keratin"},signatures:"Ling-juan Zhang",authors:[{id:"241614",title:"Dr.",name:"Lingjuan",middleName:null,surname:"Zhang",slug:"lingjuan-zhang",fullName:"Lingjuan Zhang"}]},{id:"62187",title:"Calcium and Cell Response to Heavy Metals: Can Yeast Provide an Answer?",slug:"calcium-and-cell-response-to-heavy-metals-can-yeast-provide-an-answer-",totalDownloads:1263,totalCrossrefCites:2,totalDimensionsCites:3,abstract:"Despite constant efforts to maintain a clean environment, heavy metal pollution continues to raise challenges to the industrialized world. Exposure to heavy metals is detrimental to living organisms, and it is of utmost importance that cells find rapid and efficient ways to respond to and eventually adapt to surplus metals for survival under severe stress. This chapter focuses on the attempts done so far to elucidate the calcium-mediated response to heavy metal stress using the model organism Saccharomyces cerevisiae. The possibilities to record the transient elevations of calcium within yeast cells concomitantly with the heavy metal exposure are presented, and the limitations imposed by interference between calcium and heavy metals are discussed.",book:{id:"7264",slug:"calcium-and-signal-transduction",title:"Calcium and Signal Transduction",fullTitle:"Calcium and Signal Transduction"},signatures:"Ileana Cornelia Farcasanu, Claudia Valentina Popa and Lavinia\nLiliana Ruta",authors:[{id:"203734",title:"Dr.",name:"Ileana",middleName:"Cornelia",surname:"Farcasanu",slug:"ileana-farcasanu",fullName:"Ileana Farcasanu"},{id:"203865",title:"Dr.",name:"Lavinia",middleName:null,surname:"Ruta",slug:"lavinia-ruta",fullName:"Lavinia Ruta"},{id:"255728",title:"Dr.",name:"Claudia Valentina",middleName:null,surname:"Popa",slug:"claudia-valentina-popa",fullName:"Claudia Valentina Popa"}]},{id:"61953",title:"L-Type Calcium Channels: Structure and Functions",slug:"l-type-calcium-channels-structure-and-functions",totalDownloads:2776,totalCrossrefCites:4,totalDimensionsCites:7,abstract:"Voltage-gated calcium channels (VGCCs) manage the electrical signaling of cells by allowing the selective-diffusion of calcium ions in response to the changes in the cellular membrane potential. Among the different VGCCs, the long-lasting or the L-type calcium channels (LTCCs) are prevalently expressed in a variety of cells, such as skeletal muscle, ventricular myocytes, smooth muscles and dendritic cells and forms the largest family of the VGCCs. Their wide expression pattern and significant role in diverse cellular events, including neurotransmission, cell cycle, muscular contraction, cardiac action potential and gene expression, has made these channels the major targets for drug development. In this book chapter, we aim to provide a comprehensive overview of the different VGCCs and focus on the sequence-structure–function properties of the LTCCs. 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He is currently a principal researcher in data analytics and optimisation at TECNALIA (Spain), a visiting fellow at the Basque Center for Applied Mathematics (BCAM) and a part-time lecturer at the University of the Basque Country (UPV/EHU). His research interests gravitate on the use of descriptive, prescriptive and predictive algorithms for data mining and optimization in a diverse range of application fields such as Energy, Transport, Telecommunications, Health and Industry, among others. In these fields he has published more than 240 articles, co-supervised 8 Ph.D. theses, edited 6 books, coauthored 7 patents and participated/led more than 40 research projects. 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He is currently a full professor in\nthe Department of Automation and Applied Informatics at the\nsame university. Dr. Voloşencu is the author of ten books, seven\nbook chapters, and more than 160 papers published in journals\nand conference proceedings. He has also edited twelve books and\nhas twenty-seven patents to his name. He is a manager of research grants, editor in\nchief and member of international journal editorial boards, a former plenary speaker, a member of scientific committees, and chair at international conferences. His\nresearch is in the fields of control systems, control of electric drives, fuzzy control\nsystems, neural network applications, fault detection and diagnosis, sensor network\napplications, monitoring of distributed parameter systems, and power ultrasound\napplications. 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Among them are those associated with pollution, resource extraction and overexploitation, loss of biodiversity, soil degradation, disorderly land occupation and planning, and many others. These anthropic effects could potentially be caused by any inadequate management of the environment. However, ecosystems have a resilience that makes them react to disturbances which mitigate the negative effects. It is critical to understand how ecosystems, natural and anthropized, including urban environments, respond to actions that have a negative influence and how they are managed. It is also important to establish when the limits marked by the resilience and the breaking point are achieved and when no return is possible. The main focus for the chapters is to cover the subjects such as understanding how the environment resilience works, the mechanisms involved, and how to manage them in order to improve our interactions with the environment and promote the use of adequate management practices such as those outlined in the United Nations’ Sustainable Development Goals.
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