Chapter 1: "Permanent Maxillary and Mandibular Incisors"\n
Chapter 2: "The Permanent Maxillary and Mandibular Premolar Teeth"\n
Chapter 3: "Dental Anatomical Features and Caries: A Relationship to be Investigated"\n
Chapter 4: "Anatomy Applied to Block Anaesthesia"\n
Chapter 5: "Treatment Considerations for Missing Teeth"\n
Chapter 6: "Anatomical and Functional Restoration of the Compromised Occlusion: From Theory to Materials"\n
Chapter 7: "Evaluation of the Anatomy of the Lower First Premolar"\n
Chapter 8: "A Comparative Study of the Validity and Reproducibility of Mesiodistal Tooth Size and Dental Arch with the iTero Intraoral Scanner and the Traditional Method"\n
Chapter 9: "Identification of Lower Central Incisors"\n
The book is aimed toward dentists and can also be well used in education and research.',isbn:"978-1-78923-511-1",printIsbn:"978-1-78923-510-4",pdfIsbn:"978-1-83881-247-8",doi:"10.5772/65542",price:119,priceEur:129,priceUsd:155,slug:"dental-anatomy",numberOfPages:204,isOpenForSubmission:!1,isInWos:null,hash:"445cd419d97f339f2b6514c742e6b050",bookSignature:"Bağdagül Helvacioğlu Kivanç",publishedDate:"August 1st 2018",coverURL:"https://cdn.intechopen.com/books/images_new/5814.jpg",numberOfDownloads:7237,numberOfWosCitations:0,numberOfCrossrefCitations:1,numberOfDimensionsCitations:3,hasAltmetrics:0,numberOfTotalCitations:4,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"October 4th 2016",dateEndSecondStepPublish:"October 25th 2016",dateEndThirdStepPublish:"July 16th 2017",dateEndFourthStepPublish:"August 16th 2017",dateEndFifthStepPublish:"October 16th 2017",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,editors:[{id:"178570",title:"Dr.",name:"Bağdagül",middleName:null,surname:"Helvacıoğlu Kıvanç",slug:"bagdagul-helvacioglu-kivanc",fullName:"Bağdagül Helvacıoğlu Kıvanç",profilePictureURL:"https://mts.intechopen.com/storage/users/178570/images/7646_n.jpg",biography:"Bağdagül Helvacıoğlu Kıvanç is a dentist, a teacher, a researcher and a scientist in the field of Endodontics. She was born in Zonguldak, Turkey, on February 14, 1974; she is married and has two children. She graduated in 1997 from the Ankara University, Faculty of Dentistry, Ankara, Turkey. She aquired her PhD in 2004 from the Gazi University, Faculty of Dentistry, Department of Endodontics, Ankara, Turkey, and she is still an associate professor at the same department. She has published numerous articles and a book chapter in the areas of Operative Dentistry, Esthetic Dentistry and Endodontics. She is a member of Turkish Endodontic Society and European Endodontic Society.",institutionString:null,position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"2",totalChapterViews:"0",totalEditedBooks:"1",institution:{name:"Gazi University",institutionURL:null,country:{name:"Turkey"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"174",title:"Dentistry",slug:"dentistry"}],chapters:[{id:"56461",title:"Permanent Maxillary and Mandibular Incisors",doi:"10.5772/intechopen.69542",slug:"permanent-maxillary-and-mandibular-incisors",totalDownloads:1476,totalCrossrefCites:0,totalDimensionsCites:0,signatures:"Mohammed E. Grawish, Lamyaa M. Grawish and Hala M. 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Melatonin has a major role in the regulation of circadian rhythms in non-mammalian vertebrates and forms part of their control in mammals. The present text emphasizes the positive role of exogenously administered melatonin, and its synthetic derivatives, on disrupted circadian rhythm-related dysfunctions. This is effected by resetting the clock in jet lag sufferers and those with seasonal affective disorders, insomnia, and various neurological conditions.",isbn:"978-1-83962-909-9",printIsbn:"978-1-83962-908-2",pdfIsbn:"978-1-83962-910-5",doi:"10.5772/intechopen.80180",price:119,priceEur:129,priceUsd:155,slug:"melatonin-the-hormone-of-darkness-and-its-therapeutic-potential-and-perspectives",numberOfPages:156,isOpenForSubmission:!1,hash:"bfbc5538173f11acb0f9549a85b70489",bookSignature:"Marilena Vlachou",publishedDate:"June 24th 2020",coverURL:"https://cdn.intechopen.com/books/images_new/8762.jpg",keywords:null,numberOfDownloads:1917,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:1,numberOfTotalCitations:1,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"October 28th 2019",dateEndSecondStepPublish:"February 13th 2020",dateEndThirdStepPublish:"April 13th 2020",dateEndFourthStepPublish:"July 2nd 2020",dateEndFifthStepPublish:"August 31st 2020",remainingDaysToSecondStep:"a year",secondStepPassed:!0,currentStepOfPublishingProcess:5,editedByType:"Edited by",kuFlag:!1,biosketch:null,coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"246279",title:"Associate Prof.",name:"Marilena",middleName:null,surname:"Vlachou",slug:"marilena-vlachou",fullName:"Marilena Vlachou",profilePictureURL:"https://mts.intechopen.com/storage/users/246279/images/system/246279.jpg",biography:"Marilena Vlachou is an Assistant Professor at the National and Kapodistrian University of Athens (NKUoA), Greece. She obtained her Pharmacy and PhD (Pharmaceutical Technology) degrees from the NKUoA. Just prior to obtaining her PhD she moved to the University of Rhode Island, United States, as a Visiting Research Scientist to conduct state-of-the art research on Pharmaceutical Technology techniques. Her research interests include: the formulation and in vitro release of bioactive substances from topical formulations; the efficacy and safety of formulations in skin disease therapies; the modified release of novel synthetic derivatives, with diverse activity; the investigation of the physicochemical properties of new excipients, including those of marine origin (Ulvans), and nanomaterials, with respect to their interaction with active pharmaceutical ingredients (APIs).",institutionString:"National and Kapodistrian University of Athens",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"3",totalChapterViews:"0",totalEditedBooks:"1",institution:{name:"National and Kapodistrian University of Athens",institutionURL:null,country:{name:"Greece"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"178",title:"Endocrinology",slug:"medicine-endocrinology"}],chapters:[{id:"71292",title:"Synthetic Melatonin Receptor Agonists and Antagonists",slug:"synthetic-melatonin-receptor-agonists-and-antagonists",totalDownloads:226,totalCrossrefCites:0,authors:[null]},{id:"72304",title:"Clinical Use of Melatonin in the Treatment of Sleep Disorders",slug:"clinical-use-of-melatonin-in-the-treatment-of-sleep-disorders",totalDownloads:263,totalCrossrefCites:0,authors:[null]},{id:"71288",title:"Melatonin as a Food Supplement for Sleep Disorders",slug:"melatonin-as-a-food-supplement-for-sleep-disorders",totalDownloads:235,totalCrossrefCites:0,authors:[null]},{id:"71266",title:"Melatonin for a Healthy Heart Rhythm",slug:"melatonin-for-a-healthy-heart-rhythm",totalDownloads:283,totalCrossrefCites:0,authors:[null]},{id:"71070",title:"Per Os Administered Modified-Release Solid Formulations of Melatonin: A Review of the Latest Developments Including the Design of Experiments (DoE) Approach",slug:"-em-per-os-em-administered-modified-release-solid-formulations-of-melatonin-a-review-of-the-latest-d",totalDownloads:169,totalCrossrefCites:0,authors:[null]},{id:"71009",title:"The Release Kinetics of Melatonin from Innovative Dosage Forms: The Role of the Fractal Geometry of the “Vehicle”",slug:"the-release-kinetics-of-melatonin-from-innovative-dosage-forms-the-role-of-the-fractal-geometry-of-t",totalDownloads:218,totalCrossrefCites:0,authors:[null]},{id:"71248",title:"Review of Melatonin in Horticultural Crops",slug:"review-of-melatonin-in-horticultural-crops",totalDownloads:324,totalCrossrefCites:0,authors:[null]},{id:"71664",title:"Studies on Tryptophan Metabolites in Patients of Major Monopolar Depression",slug:"studies-on-tryptophan-metabolites-in-patients-of-major-monopolar-depression",totalDownloads:201,totalCrossrefCites:0,authors:[{id:"248459",title:"Dr.",name:"Akikazu",surname:"Takada",slug:"akikazu-takada",fullName:"Akikazu Takada"}]}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"287827",firstName:"Gordan",lastName:"Tot",middleName:null,title:"Mr.",imageUrl:"https://mts.intechopen.com/storage/users/287827/images/8493_n.png",email:"gordan@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:"6581",title:"Adipose Tissue",subtitle:null,isOpenForSubmission:!1,hash:"85899eab2d8b01653e1297b168c470d7",slug:"adipose-tissue",bookSignature:"Leszek Szablewski",coverURL:"https://cdn.intechopen.com/books/images_new/6581.jpg",editedByType:"Edited by",editors:[{id:"49739",title:"Dr.",name:"Leszek",surname:"Szablewski",slug:"leszek-szablewski",fullName:"Leszek Szablewski"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7269",title:"Endocrine Disruptors",subtitle:null,isOpenForSubmission:!1,hash:"571f5c496c8b0e8db9043204fa58be2a",slug:"endocrine-disruptors",bookSignature:"Ahmed R. 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Since ancient times, humans have sought to satisfy their needs, one of which is, without a doubt, to stay alive. The fear of getting sick and dying, led man to study the organisms that surround him, discovering that the chemicals compounds present in some of them could be beneficial for treating illness. Thus; began the chemistry of the natural products; biotechnology area for human welfare. Several of these organisms produce secondary metabolites, which are part of a wide variety of natural compounds used by humans to combat diseases. Secondary metabolites are defined as organic compounds formed as bio products in organisms, not directly related to growth, development and normal reproduction of thereof. Some examples are fibers (cotton, silk, wool); fuels (oil and natural gas), and medicines (antibiotics, hormones, vaccines).
\n\t\t\tThe importance of finding and using these secondary metabolites can be justified in two ways (1) to know the natural substances that can be beneficial for man and (2) to identify the organisms that produce these substances in order to make a rational exploitation of them, because they may be the only carriers of useful compounds to combat pathogenic microbes.
\n\t\t\tMarine organisms possess an inexhaustible source of useful chemical substances for the development of new drugs; among these organisms we find marine algae that are capable of biosynthesizing a broad variety of secondary metabolites and bacteria that live in the oceans and that are crucial organisms used in biotechnology in the discovery of new compounds from marine origin.
\n\t\t\tThe discovery of new bioactive compounds necessarily involves previously diversity studies, because by knowing the type of microorganisms that reside in a certain environment, it is possible to design cultivation techniques adapted for all the microbial communities present in a certain ambience. That is why it is very important to identify the organisms that produce bioactive secondary metabolites, and to be able to structure a plan of use and preservation of those species that represent a potential source for new drug development, especially those obtained from bacteria, because of their own cultivation characteristics, have attracted attention on either a big quantity of investigators on a global scale in the search of new natural products with anticancer and antibiotic activity principally.
\n\t\tCancer is an illness that comprises more than hundred types. This disease appears when old cells are not replaced by new cells and are accumulated in a mass of tissue known as tumor (Figure 1).
\n\t\t\tPictures of immortalized cells that resemble carcinogenic cells and cells of a normal tissue (20X). (a): immortalized cells of Human Embryonic Kidney Cells (HEK293T) visualized with 20X amplification. (b): Human Coronary Artery Endothelial Cells (HCAEC) visualized with 20X amplification. (A courtesy of Aldo M.Ulloa, UCSD, School of Medicine).
The incidence of cancer increases constantly constituting an enormous challenge for health institutions, in many of the cases, the medicines used in chemotherapy treatment provoke secondary toxicity or resistance (Isnard-Bagnis et al., 2005). For all of the above, there is an urgent need to discover new anticancer compounds from natural sources. Compounds like taxol, camptothecin, vincristine and vinblastine, are obtained from superior plants (Cragg & Newman, 1999), recently some medicines obtained from marine organisms have showed promising results when administered at different cancer stages.
\n\t\t\tThe metabolic and physiological capacity that allows marine organism to survive in extreme conditions provides an enormous potential for the production of unique compounds that are not present in the terrestrial organisms. That is way marine organisms are an attractive source of compounds with pharmaceutical activity, (Faulkner, 2002). Seaweeds are recognized as one of the richest sources of new bioactive compounds of which reviews have been published recently on the biological activity of their derivative compounds (Blunt et al., 2006).
\n\t\tThe resistance to antibiotics is a phenomenon by which a microorganism stops being affected by an antimicrobial compound to which previously it was sensitive. It is a consequence of the capacity of certain microorganisms (bacteria, virus and parasites) to neutralize the effect of the medicine. The resistance can come from the mutation of the microorganism or from the acquisition of resistance genes. The infections caused by resistant microorganisms do not answer to the ordinary treatment, which result in a long illness and the risk of death (WHO, 2011).
\n\t\t\tApproximately 440 000 new cases of multiresistant tuberculosis produce at least 150 000 deaths every year. In South East Asia infections of Plasmodium falciparum that are late in disappearing after the beginning of the treatment with artemisinins are arising, which indicates resistance of the parasite to this specific medicine. Resistance has been found also to the antiretroviral medicines that are used in the treatment of the HIV’s infection (WHO, 2011).
\n\t\t\tA high percentage of the infections contracted in the hospitals are caused by very resistant bacteria, like Methicillin Resistant Staphylococcus aureus (MRSA), Enterococcus faecium and several microorganisms Gram negative resistant to Vancomycin (WHO, 2011). New mechanisms can appear that can cancel completely the ability of antimicrobial drugs to act against bacteria. This could represent the last defense against multiresistant microorganism’s strains. For example, a new β-lactamase, enzyme of the group of the carbapenemases that nowadays are named like NDM-1, gives resistance to the majority of the β-lactams medicines. The enzyme is linked to genes that are easily transferred between the common bacteria, and the infections caused by NDM-1’s producer bacteria have no treatment or, if they have it, the therapeutic options are few (WHO, 2011).
\n\t\tAccording to the World Health Organization (WHO) several factors exist that enhance the resistance to antibiotics. One problem is the lack of commitment of the government towards solving the problem, the bad definition of the responsibilities of the interested parts and the scarce participation of the consumers those results in the lack of coherent and coordinated methods to anticipate and to contain the resistance to the antimicrobial compounds. The improper and irrational use of antimicrobial drugs promotes conditions for the appearance of resistant microorganisms, which at once propagate. This happens, for example, when the patients do not take the complete treatment of a prescribed antibiotic or when the above mentioned medicine is of bad quality.
\n\t\t\tThe nonexistence or weakness of the systems of alertness determines the lack of information that can guide politicians to make recommendations and to closely continue to monitor the resistance to the existing antimicrobial compounds. Also, the scarcity of diagnosis means more medicines and vaccines for the prevention and treatment of illness, also, the shortcomings on the subject of research and development, debilitates the aptitude to combat the problem (WHO, 2011).
\n\t\t\tRight now, the WHO is focusing their efforts towards the regulation of the normatively by means of alertness, technical assistance, generation of knowledge and alliances, prevention and control of certain illnesses like tuberculosis, malaria, HIV, proper illnesses of the infancy, sexually transmitted diseases and hospitable infections; the quality, the supply and the rational use of essential medicines; the safety of the patients; and the guarantee of certified laboratories.
\n\t\t\tOn the other hand, the struggle against the resistance to the antimicrobial compounds was the issue of the World Day of the Health of 2011 (April 7). On this occasion, the WHO called to contain the spread of the resistance to the antimicrobial compounds by means of a set of politics that were recommended so that the governments can start to solve the problem (WHO, 2011).
\n\t\tMarine algae are not only used in the discovery of new drugs, they are also used extensively as food on the Asian east coast (Japan, China, Korea, Taiwan and Vietnam), Indonesia, Peru, Canada, Scandinavia, Ireland, Wales, the Philippines and Scotland, among other places. From the economic point of view the marine algae represent an important resource of food and industrial input. The Caribbean Sea coast of Colombia contains innumerable species that have an economic value and are used as human food, medicinal products, fertilizers, fuel, and play an important role in the extraction of phycocolloids and hydrocolloids (Teas, 2007). All these products have a big industrial application. In spite of the speculation on the seaweed potential as a direct source of proteins and pharmaceutical products, the demand for phycocolloids will be the factor that will influence the future development of the marine algae world resources.
\n\t\t\tMany species have been exploited, but others like the genus Sargassum and Codium have been considered to be invasive for their capacity of adaptation and their high growth rate. Due to the fact that it has been established that marine algae are a potential source for new drugs their study should become a priority. The chemical screening of all the seaweeds and their related organisms is necessary in order to establish which species can be exploited without consequences and those that must be protected.
\n\t\tIn 2010, Mexican investigators found that the marine algae Codium fragile, Sargassum muticum, Endarachne binghamiae, Centroceras clavulatum and Laurencia pacifica possess compounds that inhibit the growth of Gram negative bacteria Proteus mirabilis (Villarreal-Gómez et al., 2010), which provokes 90 % of the infections caused by Proteus. The bacteria causes the production of big levels of urease that hydrolyze the urea to ammonia increasing the pH and therefore the formation of glazing of struvite, carbonate of calcium and/or apatite, causing the formation of kidney stones.
\n\t\t\tIn the South-west coast of India, a group of scientists studied 13 groups of marine algae to evaluate the cytotoxic, larvicide, nematicide and ichthyotoxic activities on Artemia salina larvae. This Indian region is the only marine habitat with great marine algae diversity. 13 algae extracts between them Dictyota dichotoma and Hypnea pannosa showed lethal effect against the root nematode Meloidogyne javanica. D. dichotoma and Valoniopsis pachynema showed an ichthyotoxic activity. A. orientalis, Padina tetrastromatica and Centeroceras clavulatum showed activity against the urban mosquito larvae Culex quinquefasiatus (Manilal et al., 2009). Another study done in the same country, found marine algae that belonged to the family Chlorophyceae (Caulerpa racemosa and Ulva lactuca) and Rhodophyceae (Gracillaria folifera and Hypneme muciformis) that showed antibacterial activity against the Gram negative bacteria E. faecalis, K. pneumoniae and E. aerogens, as well as in the Gram positive bacteria S. aureus (Kandhasamy & Arunachalam, 2008). In a work done in the Iberian Peninsula with 82 marine algae (18 Chlorophyceae, 25 Phaeophyceae and 39 Rhodophyceae) the antibacterial and antifungal activity was analyzed to evaluate their application as natural preservatives for the cosmetic industry. The raw extracts of every taxon, prepared from fresh material as well as from lyophilized one, proved to have the opposite effect on three Gram positive bacteria, two Gram negative bacteria and yeast, by means of the agar diffusion technique. Sixty seven % of the studied seaweeds did not show antimicrobial activity against opposite the tested microorganisms. The biggest percentage of active taxon were presented in the group of the Phaeophyceae (84 %) followed by the Rhodophyceae (67 %) and finally the Chlorophyceae (44 %). The red seaweed presented the highest activity, with the widest spectrum. Inside this group, the most active species were Bonnemaisonia asparagoides, Bonnemaisonia hamifera, Asparagopsis armata and Falkenbergia rufolanosa (Bonnemaisoniales). As for the microorganisms, Bacillus cereus was the most sensitive and Pseudomona aeruginosa the most resistant. Three taxonomic groups showed seasonal change in the production of antimicrobial substances, being autumn the station with major percentage of active taxon for the Phaeophyceae and Rhodophyceae, while for the Chlorophyceae it was summer (Salvador et al., 2007).
\n\t\t\tNow a day there is a database that contains all the known natural compounds derived from marine algae, creating a crucial tool in the scientific community dedicated to the search of new useful compounds for medicinal purposes. This database provides the user with more than 3,600 released articles that describe 3,300 secondary metabolites originated from seaweeds, and it is still considered to be insufficient even though is growing every day. According to the database, Phaeophytas and Rhodophytas present significantly more quantity of bioactive compounds than Chlorophytas. The red seaweed Laurencia (Ceramiales, Rhodomelaceae) is one of the most prolific algae in the production of secondary metabolites derived from the sea. Sesquiterpenes, diterpenes, triterpernes and acetogenins (characterized by the presence of halogens atoms in their chemical structures) have been found present on this seaweed (John Davis & Vasanthi, 2011).
\n\t\tThe marine ambience is a complex ecosystem with an enormous plurality of forms of life that are associated between themselves, the most common associations found are between eukaryotic cells and microorganisms (Egan et al., 2008). The surface of all the marine eukaryotic organism are covered by microbes that live adherent to diverse communities often immersed in a matrix or forming a bilayer (Pérez–Matos et al., 2007). Also, the specificity of the guest organism also has been demonstrated in studies that show the presence of the only adherent stable communities to organisms of the same species through that they live on geographically distant regions (Webster & Bourne, 2007).
\n\t\t\tIn the recent years, the bioactive properties of marine algae and marine microorganisms have been analyzed, and in both cases positive results have been obtained. Many of the marine algae species often come accompanied by several bacterial strains which have been taken of the sea together with the algae cells, or have been the result of a contamination in the algae culture. These mixed populations that are present in the culture and in the sea, show that the bacteria use organic substances secreted by living or dead algae cell. It has been observed that many types of seaweeds present a major growth in the presence of bacteria than in their absence. Some seaweed species need vitamins for their growth and possibly the bacteria are partially responsible for the production of these substances; some of them produce antibiotics (Jasti et al., 2005; Penesyan et al., 2010).
\n\t\t\tThe number of natural products, discovered from several organism that include plants, animals and microorganisms, overcomes millions of compounds. Forty to sixty percent derives from terrestrial plants, from which twenty to twenty five % possesses bioactive properties such as antibacterial, anticancer, antifungal, antiviral and anti-inflammatory activity (Berdy, 2005).
\n\t\t\tBacteria exist only in some seaweed species, as it is the case of Leucobacter sp., collected in the Todos los Santos bay, BC. Mexico; which only was present in one out of six seaweeds analyzed (Egregia menziessi), (Villarreal-Gómez et al., 2010). This member of the Actinobacteria family, has also been associated to the nematode Caenorhabditis elegans(Muir & Tan, 2008). Micrococcus is another actinobacteria strain that has been associated only to Egregia menziessi. This strain is usually found in soil and water. It is catabolically versatile, with the skill of using unusual substrates like pyridine, herbicides, polychloric biphenyl’s and oil. It can also biodegrade many environmental pollutants (Zhuang et al., 2003). The bacterial strainKocuria palustris (Sm32), it is exclusively present in the brown seaweed Sargassum muticum that is considered to be an invasive species in many countries. This strain has industrial applications in the degradation of organic matter (Kovacs et al., 1999). The Alcaligenes found exclusively in the seaweed Endarachne binghamiae, are used for the industrial production of not standard amino acids (Madigan et al., 2005). Finally, the bacterial strain member of the genus Alteromona, was associated only with the seaweed Laurencia pacifica, generally isolated in sea water; this Proteobacteria has industrial use, since they produce polysaccharides of high molecular weight. Several of the bacterial strains phylogenetically related, have industrial application; therefore, it is necessary to study the chemical interactions seaweed - bacteria for a better understanding of the process of production of the different secondary metabolites, which produce these species.
\n\t\tThe marine microscopic communities are responsible of the change in the distribution of certain chemical elements in the sea. The autonomous aptitude of the marine organisms to produce substances biologically active that possibly accumulate, modify, kidnap and use toxins of other organisms, is a test of it (Mebs, 2000). For example, the lomaiviticins a and b, substances with antitumor potential were isolated for the first time from squids, and they contain the bacteria Micromonospora lomaivitiensis. In later experiments, this bacterium was isolated and cultivated in fermentation reactors, to finally determine that the bacteria were the real producers of lomaiviticin (He, 2001). Marine bacteria have often been considered to produce antibacterial and anticancer substances, allowing the ecological stability of the multiple marine ecosystems, the interrelations between epiphytic microorganism’s ambiences, inhibiting the rival organisms and pathogenic microbes. The sharing or competition mechanisms that are known between these microorganisms are diverse, including antibiotic production, bacteriocines, siderofores, lysosomes, proteases and even the pH alteration through the production of organic acids (Avendaño-Herrera et al., 2005).
\n\t\t\tIn recent studies done in Todos Santos Bay, B.C. to bacteria associated to the seaweed surface it was found that bacteria of the family Firmicutes, Proteobacteria and Actinobacteria produce compounds capable of inhibiting the growth of HCT-116 colorectal cancer cells (Villarreal-Gómez et al., 2010). Also, it was found that the bacteria Microbulbifer thermotolerans, and Pseudoalteromonas sp, are capable of producing biofilms and produce chemical compounds that protect them from the other protozoans. An example of these compounds is violacein, an alkaloid that it is synthesized predominantly in biofilm, it has been found that in nanomolar concentrations violacein inhibits protozoan cells and induces programmed cellular death in eukaryotic cells. This bacterial producing biofilm secretes specific chemical substances for defense purposes and contribute to the persistence of these bacterial strains in different environments and provide an ecological and evolutionary context for the discovery of bacterial metabolites against eukaryotic cells (Matz et al., 2008).
\n\t\t\t\n\t\t\t\tBacillus sp species have been found to possess chemical compounds with anticancer activity. Although this type of bacteria can grow in almost any substrate, it is possible to suggest that this species seems to have acquired the skill to synthesize compounds capable of inhibiting HCT-116 colorectal cancer cells (Villarreal-Gómez et al., 2010).
\n\t\t\tSelective response mechanisms exist against certain organisms, as shown in marine biofilms of Bacteriodetes, Planctomycetes, a,c – and d Proteobacteria, where the production of chemical substances as violacein has been observed. This compound works as a defense mechanism against certain specific predators like the protozoa consuming bacteria. This allows the successful persistence of the bacterial biofilm in several marine environments (Matz et al., 2008). Studies done in seaweed collected in the same coastal area, share similar defense mechanisms and inhibit the growth of Gram negative bacteria Proteus mirabilis and Klebsiella pneumoniae (Villarreal-Gómez et al., 2010), creating an interesting ecological and biotechnological role, and becoming a great subject for the search of marine natural products.
\n\t\t\tThe ethanolic extracts of Grateloupia doryphora, Ahnfeltiopsis durvillaei, Prionitis decipiens, Petalonia fascia and feathery Bryopsis of the central coast of Peru, presented antibacterial effect against the clinical strains Staphylococcus aureus ATCC 25923 and Enterococcus faecalis ATCC 29212 and not clinical strain Staphylococcus aureus ATCC 6633. The ethanolic extract of B. plumosa presented the biggest antibacterial effect against two strains of S. aureus, being evident in their inhibition halos, while the extract of P. fascia showed major antibacterial activity, acting on 3 mentioned above strains (Magallanes et al., 2003)
\n\t\t\tStudies done on bacteria associated with the marine worms of the Polychaetes species show a strong antimicrobial activity that can be used as a potential resource for the development of new medicines (Sunjaiy-Shankar et al., 2010).
\n\t\t\tThe following table shows some examples of bacterial strains with bioactivity and the sources where they were obtained. It is possible to appreciate the diseases that can be fought utilizing the secondary metabolites from different types of bacteria. This emphasizes the importance of microorganisms as an ideal source of bioactive compounds.
\n\t\t\tMicroorganism’s producers of bioactive substances.
Studying the diversity of bacterial species and knowing their inter and intraspecific interactions, makes the search for secondary metabolites in bacteria easier. For many years, researchers have managed to use different culturing methods that allow them to create dense bacterial populations that yield a great amount of extracts that can be used to investigate their bioactive properties.
\n\t\t\tNot only the seaweed - bacteria interactions can influence the secretion of bioactive substances, but also the interactions that exist between bacterial species that inhabit the same ecosystem. There are different types of interactions between bacterial species and other organisms; these can be positive (metabiosis and symbiosis) or negative (parasitism, predation and competition). For their high population density, nitrogen content and their relative incapability to escape from predators, the bacteria have served as food for diverse groups of organisms (Schlegel & Jannasch, 2006).
\n\t\t\tAs a rule, bacteria have been considered to be the prey of many organisms, including other bacteria species or other types of microbes. This is the case of the genes involved in producing Pilli type IV which were identified in the periplasmic strain B of Bdellovibrio bacteriovorus 109J and in the epibiotic strain Bdellovibrio sp. JSS, both Gram negative proteobacteria, which are forced predators of other Gram negative bacteria (extracellular). Using immune fluorescence microscopy the presence of the pilli was observed in the phase of cellular attack, confirming that the Pilli type IV plays a role in the invasion of other Gram negative bacteria on the part of Bdellovibrio (Qin et al., 2010).
\n\t\t\tAnother type of interaction between the bacteria is the competition. Some bacteria are eliminated by different species when the environmental resources are limited; therefore they produce compounds that impress negatively in their competitors. Generally, these antimicrobial compounds that are produced in the environment are difficult to detect, for example the bacteria incapable of producing antibiotic compounds must reproduce very fast compared to those that do. It has been found that in actinomycetes, bacterial species that have the slowest reproduction rates are the greatest producers of antibiotic compounds (Mahmoud & Koval, 2010).
\n\t\t\tIn some cases, the seaweeds dissolve organic substances that are used as nutrients by the bacteria; nevertheless some of the bacteria do not obtain nutrients this way or from any other animals. They adhere to them only for physical support and obtain their nutrients directly from the surrounding environment (Rheinheimer, 1992).
\n\t\t\tBacteria can have interactions among themselves in order to find mutual benefits. Such is the case of Escherichia coli and Proteus vulgaris; these two species can coexist in a rich in lactose and urea medium. In this case, Escherichia coli degrade lactose while Proteus vulgaris degrades urea. The final products of these degradations can be used by another organism as a carbon and nitrogen source (Rheinheimer, 1992).
\n\t\tTo this date, several bacterial identification methodologies are known to have contributed a great deal of information for the study of their diversity. Traditionally, the studies to characterize the bacterial diversity in environments are based on the assumption that the cultivation techniques allow the recovery of most of the microorganisms in a sample [39]. Nevertheless, in the database analysis of DNA sequences, it has been observed that the bacteria obtained by standard microbiological techniques represent just a limited proportion of samples from natural sources (Escalante-Lozada et al., 2004).
\n\t\t\tIt has been proposed that non-cultivable bacteria are microorganisms phylogeneticaly related to the cultivable ones, but in a physiological state that makes them recalcitrant. The explanation is that some cultivable bacteria can turn in viable, but not cultivable when exposed to adverse environmental conditions and reverse to the cultivable state after the favorable conditions for their growth are restored (McDougald et al., 1998).
\n\t\t\tTo study bacterial diversity, it is important to use molecular techniques that include the amplification of the 16S ribosomal gene using the polymerase chain reaction technique (PCR) in order to isolate and characterize their genetic material (Prieto-Davó et al., 2008).
\n\t\tTo go as far as knowing all the chemical compounds produced by bacteria extracted from different environmental samples, their isolation and cultivation, is necessary to prepare organic extracts that could be evaluated chemically and biologically. The techniques used are generally known as microbial diversity cultivation dependent techniques or traditional methods of cultivation (Joint et al., 2010). These techniques are based on the need that the microorganisms have of taking from the environment a series of compounds that are used as energy source and synthesizing the cellular constituents necessary for their survival, like C, N, S, P, Ca, K, Na, Mg, Mo, Cu and Zn. Some microorganisms need to take the light energy or to oxidize chemical compounds; others need to obtain their carbon source from CO2 or through carbonated organic compounds. Some microorganisms need a nitrogen source, for which they fix the atmospheric nitrogen, or take it from NH4\n\t\t\t\t+. Others are capable of reducing NO3\n\t\t\t\t- or NO2\n\t\t\t\t- to use it as an ammonium source, or, to use free amino acids. All these characteristics, as well as temperature, pH and salinity, have demonstrated to create a propitious ambient, for the isolation and purification of microorganisms originated from environmental samples (Rheinheimer, 1992).
\n\t\t\tTherefore, the collected environmental samples are exposed to different conditions and forms of cultivation that help to obtaining microorganisms capable of adapting to the established conditions. This allows to study the microorganism’s diversity, as well as to cultivate them from different environmental samples.
\n\t\tThe molecular characterization of bacteria has had an enormous impact on the safety of microbiological industrial processes like the bio pharmacology and food industry as well as public health, since it establishes the source of contamination in a much more precise form and therefore to identify the strains involved in the process and to establish their trajectory.
\n\t\t\tThe most used gene for bacterial molecular identification is the 16S ribosomal RNA gene, nevertheless, there have also been used other genes that can codify for 5S ribosomal proteins, and 23 S ribosomal RNA.
\n\t\t\tThe ribosomes and ribonucleic acids (ribosomal RNA) are proteic complexes with the only function of synthesizing proteins. The ribosomes structure is preserved between the different life kingdoms (Plantae, fungi, animaleae, etc.). In the prokaryotes, the ribosomes are composed by two subunits: a big 50S, and small 30S subunits. The subunit 50S contains a 23S, one 5S rRNA and more than 30 different proteins. The subunit 30S contains one 16S rRNA and 20 additional proteins. Since the rRNA gene function is limited by the structure, certain regions in the rRNA genes that are in contact with other components in the ribosome must be preserved; the sequences between the preserved regions have major mutation valuations. The preserved regions are useful to determine distant relations (genus), while the regions with higher mutation rate or variable regions are useful to distinguish organism closely related (species) (Eickbush & Eickbush, 2007). These characteristics of rRNA genes are excellent molecular chronometers for phylogenetic analysis and taxonomic classification of cellular organisms.
\n\t\t\tScheme of Bacterial 16S ribosomal RNA that shows the variable secuences (V1 to V9). The variable regions area used to characterized bacteria by species level, and the constant region is used to correlated the genus.
In general, the phylogenetic trees based on the 16S and 23S rRNA genes can be used in parallel (De Rijk et al., 1995), while the 5S rRNA gene it is considered not to contain sufficiently long sequences to do significant statistical comparisons. The detailed phylogenetic studies based on the genetic sequences 16S and 23S provide comparisons in three primary domains Bacteria, Archaea and Eukarya (Woese et al., 1990). Nevertheless, one of the disadvantages of the 23S rRNA gene on 16S rRNA in the phylogenetic studies is the rare use of the taxonomic classification for the absence of primers that amplify a considerable length sequence and the difficulty of genes sequences too long for the current technologies.
\n\t\t\tPreviously the 16S rRNA and 23S rRNA genes have been compared and the results show that the 16S rRNA gene has more closely related typical sequences, with major length, insertions and/or deletions and possibly a better phylogenetic resolution for their high genetic change (Blunt et al., 2005). Nevertheless, recent studies indicated that the 23S rRNA gene also contains conserved regions for the design of primers capable of amplifying a wider length with one grade similar to the primers used in the of 16S rRNA genes (Hunt et al., 2006; Pei et al., 2009).
\n\t\t\tMolecular methods of characterization based on the amplification of fragments belonging to the 16S ribosomal RNA gene through automated techniques constitute a rapid, trustworthy and simple method of molecular genotyped bacterial and fungoid strains. The aptitude to identify microorganisms at species level and at the same time to establish a comparative analysis of the different analyzed strains constitutes a time saving way to identify the food pollutant potential and therefore to eradicate the pollutant focus or to facilitate the recognition of the producing strains of some bioactive metabolite capable of inhibiting the growth of other bacterial strains or carcinogenic cells.
\n\t\t\tThe 16 S ribosomal gene (16S rRNA) is constituted by a region preserved through evolution, the mutations in this gene can usually be tolerated, since these mutations would only affect ribosomal RNA, nevertheless, the number of mutations are not completely well-known, the regions that are affected by them are met like “hot commercials” which present a considerable number of mutations, these areas are not the same in all species. The 16S rRNA gene mutations can affect the susceptibility to antimicrobial agents which can be an indicator to distinguish the phenotypic resistance to these agents. Nevertheless these characteristics do not affect the gene use for taxonomical identification at the genus and species level. 16S rRNA possesses a length of 1, 550 bp and it is composed by variable regions and conserved regions, with enough interspecific polymorphisms to provide a valid statistical characterization. The primers that are usually designed for the amplification of this gene, are based on the complementary chain of the conserved regions in the beginning of the gene at about 540 bp or at the end of the sequence around 1,550 bp and the sequence of the variable region is used for taxonomic comparison intentions, making 1500bp the minimum length that should be used to compare DNA sequences. The 16S rRNA gene sequence has been determined for a big number of strains; these sequences are included in the biggest database of nucleotides known as GenBank. This database contains more than 20 million sequences of which more than 90, 000 correspond to the 16S rRNA gene.. In general, the comparison of the 16S rRNA gene sequences allows the differentiation between organisms at genus level in most bacteria phyla, also classifies the strains at multiple levels, including species and sub species (Clarridge III, 2004).
\n\t\tTo do a search of bioactive compounds from marine algae the following protocol is proposed:
\n\t\t\t\n\t\t\t\ta. Isolation of the bacterial strains from the seaweed surface\n\t\t\t
\n\t\t\tAfter being collected, the seaweed is placed in an Erlenmeyer flask and is rinsed with distilled water; a small sample from the flask is taken with a sterile swab and is inoculated in a general media that allows the growth of most of the bacteria present in the seaweed surface. Then the media with inoculate is incubated for periods of 24 to 48 hours at 25 oC, until the developing colonies start to emerge. Later the bacteria will be purified up to the third generation to assure the integrity of pure colonies and finally it is important to take one of the colonies of every purified strain and cryopreserved it in 15 % glycerol at -70 oC.
\n\t\t\t\n\t\t\t\tb. Macroscopic morphology and Gram Stain\n\t\t\t
\n\t\t\tThe pure colonies are examined macroscopically evaluating their characteristics such as size, form, elevation, margin, color, type of surface, thickness, consistency, smell and pigments production. Consecutively microscopic classification of the bacterial strains, such as the Gram stain, is necessary (Gram, 1884). The Gram stain is a technique of bacterial characterization based on the chemical composition of the cellular wall of the bacteria. The Gram positive bacteria present a cytoplasmic membrane, have a thick peptidoglycan layer, contain teicoics acids and lipoteicoics that serve as chelating agents and certain type of adhesions; the bacteria representative of this group are Firmicutes and Actinobacteria, which includes many well-known genus like Bacillus, Listeria, Staphylococcus, Streptococcus, Enterococcus, and Clostridium.
\n\t\t\tProposed methodology for the search of new bioactive compounds in bacteria associated with the surface of marine algae.
On the other hand, the Gram negative bacteria contain a thin cellular wall of peptidoglycan and an external membrane that covers their cellular wall. The external membrane contains diverse proteins, like purines or channels protein that allow the path of certain substances. Also they present a structure of lipopolysaccharides (LPS). The bacteria that predominate in this group are Proteobacteria, including to Escherichia coli, Salmonella and other enteric bacteria like Pseudomonas, Moraxella, Helicobacter, Stenotrophomonas, Bdellovibrio, acetic acid bacteria, Legionnaire\'s disease and the proteobacteria alpha like Wolbachia among others (Madigan et al., 2005).
\n\t\t\tPrevious studies done with bacterioplacton demonstrated that most of the marine bacteria are Gram negative; but in recent studies with marine sediments evidence has shown that most of the bacteria that conform the marine sediments seem to be Gram positive (Gontang et al., 2007).
\n\t\t\tIn studies made to the surface of the marine alga Monostroma undulatum (Gallardo et al., 2004) Gram negative bacteria belonging to the genus: Vibrio (20 %), inactive E. coli (18 %), Flavobacteria (11 %), Flexibacter (9 %), Moraxella (9 %), Pseudomona (9 %), Aeromonas (2 %), Acinetobacter (2 %), Cotophaga (2 %), Photobacteria (2 %) and Alteromonas (2 %); predominated and only one Gram positive was found, Staphylococcus.
\n\t\t\t\n\t\t\t\ta. Phylogenetic analysis\n\t\t\t
\n\t\t\tSince bacterial identification was mentioned previously and bacterial characterization with molecular techniques is crucial for the achievement of phylogenetic studies that allow to have an account of the cultivable bacterial species that are present in a similar community, there are several investigators who agree with the idea, that bacteria with closely related DNA sequences produce very similar compounds. If this is true, just by knowing the bacterial ecology of a certain area it will be possible to predict the types of compounds that could be isolated from the bacteria present in that particular area.
\n\t\t\tThe DNA sequences can be analyzed using the BLAST database (Basic Local Alignment Search Tool) that is a GeneBank database integral function (Altschul et al., 1999). To align the existing sequences between themselves it is possible to use the Clustal X (Staley & Ta, 1985) and Bioedit programs (Hall & Brown, 2001) for manual alignments. For the phylogenetic tree construction it is possible to use the following indications: Bootstrap test of phylogeny (1000 repetitions), p-distance joining neighbor, using the MEGA4 program (Tamura et al., 2007) using segment sequences of up to 1500 bp.
\n\t\t\t\n\t\t\t\tb. Biological assays and mean lethal dose DL\n\t\t\t\t\n\t\t\t\t\t50\n\t\t\t\t\n\t\t\t
\n\t\t\tA bioassay can be defined as any test that involves living organisms. With them, it is possible to evaluate the effects of any substances or material in terms of the biological answer they produce. The main target in this type of analysis is to evaluate the level of stimulus that is necessary to obtain a response in a group of individuals of a population. The level of stimulus that causes response in 50 % of the individuals under study is an important parameter of characterization denoted like average lethal dose (DL50). The amount of time during which the stimulus is exhibited, must be specified, for example, 24 hours DL50, this in order to compare and to estimate the relative potency of the stimulus. For the DL50determination of the first step is securing the % of survival cell for every analyte and every target, the mortalities is corrected by Abbott´s formula (Abbott, 1925).
\n\t\t\tWhere:
\n\t\t\t\n\t\t\t\tM = Mortality.
\n\t\t\t\n\t\t\t\tme = optical density in the extract.
\n\t\t\t\n\t\t\t\tmb = optical density in the target.
\n\t\t\tWith the mortalities corrected, the obtained information is introduced in the software “BioStat 2008” (http://www.analystsoft.com) using Probit analysis to obtain the DL50values (STATPLUS, 2008).
\n\t\tNatural products are a very important resource for the elaboration of medicines. Although a big number of plants, microbes and marine resources have been evaluated in the search of new bioactive compounds, it turns out to be insufficient and it is necessary and important to continue with the search of new secondary metabolites, especially those that are endophytes microorganisms of seaweed. The bad use given to antibiotics has resulted in the development of bacteria strains that are resistant to many of the known drugs. This situation has lead to a forced search for new antibiotic compounds, being the seabed a propitious site for exploration and future drug development. Also, the treatment with chemotherapy for the diverse causes of different types of cancer that at present today, appears effective, so the investigation becomes necessary in the chemistry of the natural products. The methods of bacterial culture and identification have become very promising especially, those done through molecular techniques, by which is possible to identify a strain up to species and sometimes at subspecies level. The diverse relationships that exist between microorganisms and their guests provoke that bacterial compounds can eventually be used as a source of new drugs for human well-being.
\n\t\t\t\n\t\t\t\tFuture work in Drug Discovery\n\t\t\t
\n\t\t\tThe strategies for drug discovery has been evolving constantly, today researchers do not conform with the finding of new and potent metabolites, now and for future days it has become important to do phylogenetic studies, structure elucidation of chemical compounds, bioinformatics approaches, genomics, proteomics, reverse pharmacology and so on., One fundamental field that has to be developed is the improvement of more efficient culture media, because we need to culture all the strains of bacteria to be able to evaluate and separate its compounds, in the meanwhile, we can identify the genes of cultivable and non-cultivable bacteria by molecular techniques to compare and try to demonstrate that the strains of bacteria with very similar DNA sequences have equally similar metabolites and culture requirements, if this is true, before we start screening for drug discovery purposes, researchers must do phylogenetic studies of bacterial population of a certain determinate area and decide which strains cultivate and which not, these strategies will make the drug discovery process less expensive and faster.
\n\t\t\tMethodological improvements studies based on the characterization of the extracellular polymeric substance produced by marine microorganisms and a better understanding of host-microbe interactions, should be us to provide further insight into the adaptive strategies against microbial pathogens and establish the extent to which secondary metabolites regulate microbial interactions.
\n\t\t\tThe new soft ionization methods: Matrix-Assisted Laser Desorption Ionization (MALDI) and Electrospray Ionization (EI) are the recent approach used in a variety of new and innovative Mass Spectrometric (MS) applications. With them, is easier to analyze surfaces, they are tolerant to impurities and do not require extensive sample preparations. A sensitive and precise Mass Spectrometric approach like Desorption Electrospray Ionization (DESI) should be used to measure the physical location and quantities of natural products on biological tissue surfaces, cells or even complex mixed-species assemblages. These MS imaging techniques known as “molecular eyes” are very precise and represent the last technological advance used to locate natural products in biological tissues (Esquenazi et al., 2009) allowing the study of the interface between the confluence of natural products chemistry, biology and ecology.
\n\t\t\tBioinformatics is the part of molecular biology that involves working with biological data, typically using computers, with the goal of enabling and accelerating biological research. Bioinformatics comprises a wide range of activities: data capture, automated recording of experimental results; data storage and access, using a multitude of databases and query tools; data analysis; and visualization of raw data and analytical results (Pollock & Safer, 2001).
\n\t\t\tToday, many recently developed or discovered drugs with antibacterial and anticancer activity fail in clinical trials because of inefficiency for the anticipated indication or unexpected toxicity (Kola & Landis, 2004). Apparently, it remains hard to establish a clear link between antagonism or organism of a specific target and its influence in human illness and its target associated toxicity.
\n\t\t\tA significant cause for these high attrition rates is the often misjudged complexity of protein function in higher order organisms, in which, abundant protein-protein interactions, feedback loops and redundancies play a role. The collection of recognized pathways that can be found in public databases and commercial tools do not effectively address these issues because they are mainly a reflection of experimental data that are obtained from isolated cell lines and tissues. They address typically, the signaling events that lead to binding of transcription factors to the DNA, but do not detail the pleiotropic effects that arise downstream from the induced transcriptional program, which are most important in provoking the systems response to the signaling events and may determine, the capacity and toxicity of a drug (Pollock & Safer, 2001).
\n\t\t\tMost comparative genomics tools are intended at studying conservation of single genes or gene families, whereas computationally tools address orthologous biology, i.e. conservation of the entire pathways in which the target is involved, are unusual. This truly obstructs the output and success of translational investigation from pre-clinical to clinical studies (Pollock & Safer, 2001).
\n\t\t\tThe developing of bioinformatics tools that addresses the above problems will allow for quicker and better experiments aimed at evaluating multiple targets and drugs for further clinical development. This will be a first step to reduce the high attrition rates associated with drug development (Pollock & Safer, 2001).
\n\t\t\tClinical events or phenomena not reported previously following the administration of a known or new drug can offer valuable perceptions for drug development. Natural products have provided many such unexpected bedside interpretations. Researches in genomics, proteomics and metabolomics have stimulated the discovery of many new molecules, which are yet to be tracked for their drug-like activities. A new discipline called Reverse Pharmacology (RP) has been designed to decrease costs, time and toxicity.
\n\t\t\tThe scope of reverse pharmacology is to understand the mechanisms of action at multiple levels of biology and to optimize safety, efficacy and acceptability of the leads in natural products based on relevant science in this approach, as the candidate travels a reverse path from ‘clinics to laboratory’ rather than classical “laboratory to clinics”. Actual humans are used as the ultimate model and in-depth investigation of the effects of drugs and the nature of disease progression is becoming ever more feasible because of advances in clinical biomarkers and systems biology. This articulates both structure of the system and components to play indispensable role forming symbiotic state of the whole system (Patwardhan et al., 2008).
\n\t\tThe authors thank A. Prieto-Davo and P.R Jensen (CMBB,U.C.S.D.) for their collaboration, A. Licea for his help in carrying out the bioassays (CICESE),M. Ritchie for facilitating the bacterial pathogens used, R. Aguilar (†) for assistance in the seaweed identification, A. Moreno and L. Pérez for photography and Illustration, respectively and A.M. Iñiguez-Martínez for reviewing the manuscript.
\n\t\tNeurotoxins are toxic substances that destroy the nervous system. Depending on their origin, it could be divided into endogenous or exogenous neurotoxins. Endogenous is the neurotoxin produced by the human body itself, while exogenous one comes from the surrounding environment. Neurotoxins can damage neurons, nerve fibers, glias, and myelin, causing the atrophy of nerve fibers and neurons, or demyelination, which in turn affects neural circuits and functions. Ultimately, defect in nerve system affects the physiological homeostasis of human body, which results in corresponding signs and symptoms of poisoning. Macro-manifestations of neurotoxin exposure may associate a wide range of central nervous system impairments such as cognitive deficit [1], memory impairment [2], epilepsy, and dementia [3, 4].
Exogenous toxins are foreign synthetic by name, and common exogenous neurotoxins include metal neurotoxins (e.g., lead), microbial neurotoxins (e.g., botulinum), biotoxins (e.g., tetrodotoxin), and chemical toxoids (e.g., ethanol). Different types of toxins have their different mechanisms of action in nervous system: (1) metal neurotoxins, such as lead and aluminum, usually migrate to the brain through the blood circulation by destroying the structure of or inhibition of the blood-brain barrier (BBB) [5]. Once it penetrates the BBB and reaches the brain, it can cause damage to brain and thus the emergence of diseases such as learning disabilities [6, 7], disorders in motor coordination [8, 9], and Alzheimer’s disease [10]. At present, these metal neurotoxins are still widely used in food preparation, like in the packaging factory. Furthermore, due to the environmental pollution, these substances are also widely existing in the food chain [11]. (2) Microbial neurotoxins are produced by microorganisms, mostly from bacteria, such as botulinum, tetanus toxin, and lipopolysaccharide (LPS). The main mechanism of microbial neurotoxins that disturb the nervous system is inhibiting the communication of neurons. Microbial neurotoxins prohibit the release of neurotransmitters from synaptic vesicle [12, 13], thereby terminating nerve messages, which may lead to a decrease in muscle tension [14, 15], muscle atrophy [16], and paralysis [17, 18]. If it affected the respiratory muscles, it could cause asphyxiation and death [19, 20]. (3) Bio-neurotoxins come from organisms that produce these toxins as tetrodotoxin existed inside pufferfish\'s skin and gastrointestinal tract, snake venom produced by snakes, and chlorotoxin produced by scorpion. Some of which reduce the permeability of the ion channels in neurons and thus decrease neuronal communication. Different bio-neurotoxins can target different ion channels. For example, the tetrodotoxin is specific to the sodium ion channel [21], while the conotoxin produced by the conch is specific to the calcium ion channel [22]. The other bio-neurotoxins may not affect ion channel but have impact on neurotransmitter gated channel, like bungarotoxin, a type of snake venom [23]. (4) Chemical neurotoxins are a class of toxins with a broader mechanism of action. For example, ethanol has been shown to induce nervous system damage and affect the body in various ways: studies show that ethanol can alter the composition of nerve cell membranes [24], inhibiting the activation of NMDA receptors [25], causing the imbalance of cellular calcium ion concentration [26], facilitating the mitochondrial dysfunction [27], and increasing the oxidative stress inside the neurons [28], and that destroy the nervous system, leading to brain atrophy, encephalitis, neurodegeneration, cognitive decline, developmental disorders, and other neurological diseases [29, 30].
Endogenous neurotoxins, such as nitric oxide and glutamate, originate in the body and usually have their typical physiological role and function in the body. When the concentration of these endogenous compounds becomes higher, it can lead to dangerous effects: (1) glutamate is the primary neurotransmitter of the nervous system, accomplishing the chemical transmission in synapses. The normal concentration of glutamate is responsible for the regular performance of neurons. One of the most critical uses in nervous system is an excitatory neurotransmitter, which is related to the long-term potentiation in memory and learning. High concentrations of glutamate become toxic to the neurons by increasing the permeability of calcium ions. It leads to an increase in cellular calcium concentration, then over-activates the calcium-associated enzymes, and eventually results in neuronal swelling and cellular death. This phenomenon is known as excitotoxicity. Studies have linked this mechanism to many neurological disorders, such as Huntington\'s disease, epilepsy, and stroke [31, 32]. (2) Nitric oxide (NO) is a secondary messenger synthesized by neural nitric oxide and commonly used in neurons. It regulates synaptic plasticity of the nervous system, smooth muscle relaxation in nerve and vascular system, and neurovascular dilation [33, 34]. Abnormal concentration of NO is associated with asthma, schizophrenia, and Huntington’s disease [35, 36, 37]. The neurotoxicity of NO is based on glutamate-induced excitotoxicity. NO is response to glutamate-mediated NMDA activation, which is produced by calcium-dependent signaling. An elevated rate of glutamate excitotoxicity could lead to an increase in neuronal NO level. Over-dose of NO can also increase oxidative stress, which further induces DNA damage and apoptosis [38]. Therefore, an abnormal level of NO inside the nervous system can produce significant neuronal toxic effects.
Common antidotes, such as antioxidants and antitoxins, can effectively reduce nerve damage induced by neurotoxins: (1) Antitoxin or antiserum is an antidote that uses antibodies to neutralize specific action of the toxins. Antitoxin is produced by individual animals, plants, or bacteria that are responded to toxin exposure. Antitoxins are made in organisms and can be injected into other organisms, including humans. Its most common use in the human body is antivenom [39]. (2) Antioxidants are compounds that inhibit oxidative stress, such as glutathione or ascorbic acid (vitamin C). If neurotoxins cause oxidative stress, antioxidants can be used to reduce the toxic reactions and side effects [40, 41].
Brain is the computational core of our nervous system and is a place where animals can process and coherent the stimulus gathering from outside or internal surroundings and then sending out the response. It is responsible for many higher-order functions such as coordination of movement, learning and memory, and language and speech. Therefore, a well and the fully developed brain is essential to regulate these functions. Many intrinsic and external factors are required for brain development. Intrinsic factors are like hormones or regular development-associated gene expression, and external factors are like essential nutrients. Abnormal impact on these factors results in brain malformation and malfunction. For example, abnormal expression of thyroid hormone affects cerebellar development, motor performance, and severe anxiety [42, 43]; dysfunction of cyclin-dependent kinase 5 and insulin-like growth factor-I results in neurological disorders and neurodegenerative diseases [44, 45]; iron deficiency in early life causes irreversible effect on behavioral and neural development [43, 46]; and (n-3) fatty acid plays a role in neurogenesis, neurotransmission, and protection against oxidative stress in whole life span [47].
During cerebral development, brain cells go through the process that includes replication and migration. The replication of the brain cells is also called neurogenesis, which starts from the 8th week of gestation in humans and the 10th day in rodents [48]. Neurogenesis is the formation of neurons from neural stem cells near the area of lateral ventricles occurring during embryonic development and is responsible for producing all the various types of neurons of the brain. Neuronal migration mainly happens between the 12th and 24th week of gestation in humans and the 11th and 16th day in rodents [49, 50]. Neurons pass the subplate and migrate into the cortical plate along the radial glial process starting from the subventricular zone with an inside out pattern that forms six-layered neocortical laminae [51], also known as cortex. We may look at these processes in detail through rodent studies: neuronal progenitor cells (NPCs) form the earliest cortical neurons and lie inside the preplate on the 10.5th day of pregnancy, which is divided into the superficial marginal zone, the cortical plate, and the subplate. The radial glial cells (RGCs) are also generated from NPCs at around 11th to 12th day of gestation, which owns the unique morphology with their soma inside the ventricular zone, their short processes extending the apical side of ventricle, and their long processes elongating to basal lamina. Newly formed neurons use the processes of RGCs as guiding railroad to migrate from the ventricular zone toward their final destination inside the cortical plate. Early-born neurons give rise to layers I, V, and VI, and later-born neurons migrate from past layers V and VI to the other layers (layers II to IV) of the cortical plate [52]. This is also known as in-side-out migration.
Different cortical layers have different functions; for example, the neurons of the 6th layer remarkably express the T-box brain 1 (Tbr1) protein, which regulates cell migration and differentiation during embryonic development [53, 54, 55], and also is responsible for the connection between cortex and thalamus in the developed brain [56]. The cortical neurons of the 5th layer mainly express COUP-TF-interacting protein 2 (Ctip2; also known as Bcl11b), a C2H2 zinc finger transcription factor, and link the cortex with brain stem and spinal cord [57]. Special AT-rich sequence-binding protein 2 (SATB2) also known as DNA-binding protein SATB2 is highly expressed in the neurons with layers II and III, which is associated with neuronal morphogenesis [58]. In adult brain, the neurons in layers II and III mediate communication across cortical regions and with the amygdala [59, 60].
After the nerve cells migrate to the destination, they begin to develop neurites in order to form synapses, which communicate with other brain cells. Neurites are composed of axons and dendrites. Axons transmit messages, and dendrites receive them. Both of which shape neural circuits. Most of the topics discussed are dendritic patterns. Dendrites are highly branched bush-like cellular extensions that mediate the enormous majority of presynaptic and environmental inputs, which regulate neuronal communication. Dendritic patterning is critical for proper neuronal function and has served as the basis for the classification of neuronal subtypes [61, 62]. The development of dendrite is a complicated multistep process. First, neurites initiate and form a structure called lamellipodia. Axons outgrow from minor processes followed by the outgrowth of dendrites. Dendrites then start to branch and form dendritic spines. After the process of pruning, dendritic branches are fully matured, and synapses are formed [62, 63, 64]. These steps are regulated by intrinsic genetic signals and extracellular cues [64]. Disruption of these pathways results in abnormal development in dendrite patterning that sequentially affects communication between neurons, which further leads to disruption of neuronal circuitry, and finally, whole nervous system breaks down. For example, mutation in the human neural cell adhesion molecule L1 and Neuro-p24, a membranous protein, affects neurites’ outgrowth and extension [65, 66]. Many well-known neurodegenerative diseases, such as Rett syndrome (RS) and autism, are genetic defects with well-defined anomalies in dendritic patterning [67, 68]. Hence, the integrity of dendrite morphology is crucial for maintaining normal function of brain circuitry and neuronal networks.
Gene is the main intrinsic factor that affects brain development, which contained highly programmed information related to neurogenesis and migration. These genetic factors include integration of reelin, Lis-1, and doublecortin [50, 69, 70]. The most studied genes are those encoding the dopamine inactivator catechol-O-methyl transferase (COMT), neurotrophin, brain-derived neurotrophic factor (BDNF), the schizophrenia candidate gene neuregulin (NRG1), and the serotonin transporter (5-HTT), for example, take neurotrophins. Neurotrophins play an essential role in mediating neuronal survival and brain development. There are four types of neurotrophins: neuronal growth factor [71], brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3), and neurotrophin 4/5 (NT-4/5). In their mature forms, these neurotrophins can bind as homodimers to specific tropomycin-related kinase (Trk) receptors. TrkA binds to NGF, TrkB binds to BDNF and NT-4/5, and TrkC binds to NT-3. Trk receptors are part of a group of receptor tyrosine kinases. Ligand binding results in dimerization of the receptor. The dimerized receptors phosphorylate several conserved tyrosine residues on one another [72, 73, 74]. This allows for proteins containing phosphotyrosine binding (PTB) or Src homology (SH2) domains to dock, and this docking activates intracellular signaling cascades that include Ras-Raf-Erk, PI3 kinase-Akt, PLC-γ-Ca2+, NF-κB, and some protein kinase C pathways [73]. Neurotrophins exist in two states: proteolytically processed, which is the mature form and can bind the various Trk receptors, or unprocessed, which allows them to bind with high affinity to p75 neurotrophin receptor (NTR). Processed neurotrophins can still bind to p75NTR but with much lower affinity than to the Trk receptors. Binding of processed or unprocessed neurotrophins to p75 may elicit several responses including cell death [72, 73]. Recent studies also show that extracellular stimuli (NGF, BDNF, and epidermal growth factor) induced Rac activity, which is involved in neurotrophin-derived signaling and neuronal migration [75]. Deficiency in these intrinsic regulators disrupts neuronal migration and cortical laminar organization [76, 77], causing morphological abnormalities, such as schizencephaly, porencephaly, lissencephaly, macrogyria, and microgyria [78]. These lead to psychiatric and neurological disorders [69, 79, 80].
The blood-brain barrier (BBB) is a crucial example of protection against toxins and other adverse compounds reaching the brain. BBB of the developing brain is not fully formed at the beginning of gestation, and therefore, fetal will be more vulnerable to the neurotoxins [81, 82]. For example, long-term exposure to nicotine during pregnancy directly affects acetylcholine systems and brain cell replication and differentiation, which consequently results in behavioral deficits of the offspring [83, 84]; prenatal, postnatal, and adolescent administration of alcohol affects gene expression (e.g. c-fos), causes abnormalities of brain structure and function, and finally disrupts brain development permanently and irreversibly [85, 86].
In the current industrialized society, atypical neurotoxins may also have hidden influence on normal brain development. The impacts of these agents on the processes of brain development are diverse and the productive time extends from prenatal to adolescent period [85]. Prenatal exposure to carbofuran, a carbamate pesticide, decreases nestin expression, histone-H3 phosphorylation, and the number of glial fibrillary acidic protein and SOX-2 co-labeled cells, which further leads to neurodegeneration and cognitive deficits in offspring [87]; Maternal application of dicholoacetonitrile (a disinfectant in our drinking water), benzyl benzoate (a antiparasitic insecticide), and trimethyltin (a stabilizer for plastics in paints) may induce oxidative stress and then result in neurodegeneration in fetal brain [88, 89, 90]; maternal infection is associated with the increased levels of proinflammatory cytokines in the amniotic fluid [91], umbilical cord plasma [92], and cerebral palsy [93, 94] as well as neurodevelopmental disorders such as schizophrenia [95] and autism spectrum disorders [96]. The findings in experimental models of maternal infection manifest the role of inflammatory response in the alteration of fetal neuronal morphology [97], astrocytosis, ventriculomegaly, changes in oligodendrocyte precursors [98], reduction of oligodendrocyte number, hypomyelination of brain [99] and a decrease of dopaminergic and serotoninergic neurons in the offspring [100], all of which are capable of leading to brain formative deficits. Indeed, maternal infection induces changes in brain developmental events, including neurogenesis, myelination, synaptogenesis as well as cell migration [101, 102]. Neuroinflammation has been reported to be highly associated with numbers of neurological and pathological diseases, such as cerebral palsy [94], schizophrenia [95], and autism spectrum disorders [96].
However, the precise molecular mechanisms of these exogenous agents that cause abnormal brain development largely remain unknown. Moreover, there are other possible harmful candidates needed to reveal. For example, monocyclic aromatic amines (MAAs) are a group of chemicals ubiquitously present in the environment. Exposure assessments indicate that most individuals experience lifelong exposure to these compounds from several sources, such as occupational exposure via tobacco smoke, herbicides, or hair dye, which are considerable in causing bladder cancer [103]. In vivo evidence has demonstrated the carcinogenic potential of most alkyl aniline compounds. For examples, 2,6-dimethylaniline (DMA) is responsible for nasal carcinogenesis [104]; Gan and colleagues indicated that 2,6-DMA, 3,5-dimethylaniline (3,5-DMA), and 3-ethylaniline (3-EA) are strongly associated with bladder cancers [105]. Currently, only 2,6-DMA is categorized as a possible human carcinogen by IARC [106]. However, the threats of other alkyl anilines cannot be neglected that 2,6-DMA, 3,5-DMA, and 3-EA can be metabolized as electrophilic intermediates, which further bind to DNA and form adducts. Skipper and colleagues report that three alkyl anilines can be metabolized as electrophilic intermediates and induce the production of DNA adducts, followed by attacking their putative targets, like bladder. Moreover, their results indicated that the adduct levels were the highest in animals given 3,5-DMA and the lowest in that given 3-EA. Furthermore, 3,5-DMA has been indicated not only to play a significant role in the etiology of bladder cancer in humans but study using in vivo experiments also strongly suggested that DNA adducts formed by 3,5-DMA might account for its presumptive activity [107]. A recent study additionally proves that 2,6-DMA and 3,5-DMA cause a single base-pair transition in the guanine-hypoxanthine phosphoribosyltransferase (gpt) gene in an in vitro model [108]. MMAs are activated through cytochrome P450-catalyzed oxidation of the amino group, followed by extensive esterification of N-hydroxylamine and heterolysis of the N▬O bond to produce a reactive nitrenium ion. The ion then interacts with DNA base and forms covalent adducts [108, 109]. Furthermore, the other major product of hydroxylation, aminophenol, also has the ability to damage DNA by electrophilic attack at nucleophilic DNA bases that lead to mutagenesis and carcinogenesis. Aminophenol becomes electrophilic upon 2-electron oxidation to quinine imines, followed by Michael addition reactions as well as nucleophilic addition at the keto and imino carbon centers. Finally, DNA adducts are formed [103, 110]. Moreover, quinines react directly with proteins through thiol addition. Thioether addition products are responsible for the production of reactive oxygen species (ROS) [111, 112]. Accordingly, oxidation of alkyl anilines is generally regarded as a critical bioactivation. Indeed, studies have shown that the major metabolite of 3,5-DMA and 3,5-dimethylaminophenol (3,5-DMAP) is responsible for the ROS production, which contributes to the apoptotic cytotoxicity in mammalian cell lines [113, 114]. Furthermore, a recent in vivo study has shown that 3,5-DMA-induced ROS production disrupts the dendrites patterning of the cortical cells and causes the abnormal cortical layer distribution in developing fetal brain [115].
The most popular topic of air pollution, fine particular matter such as PM2.5, has been reported to have massive impact on neurodevelopment. A study displays that PM2.5 can enter the maternal amniotic fluid system by inhalation and directly causes the delay in brain development of the fetal rat. Besides, microarray data demonstrated that PM2.5 mainly increases the risk of neurological diseases in the offspring, such as Alzheimer’s disease, epilepsy, autism, learning and memory disorders, and emotional control disorders [116]. Up-to-date study shows that PM2.5 increases the number of white blood cell, upregulates the inflammatory response, induces the memory impairment, and declines in dendritic branches of the hippocampi of the offspring [117].
The human brain is the most complex system in biology, and its function depends on the various connections of nerve cells or the interaction between brain regions. Besides, there is growing evidence that many mental and neurological disorders are associated with neurodevelopmental abnormalities, which, according to epidemiological statistics, include autism, attention deficit hyperactivity disorder, dyslexia, and other cognitive impairments affecting millions of children worldwide [118, 119]. During the fetal and infant periods, they are most susceptible to environmental neurotoxin, which can cause permanent brain damage during these sensitive developmental stages [82]. Classical neurotoxins, such as heavy metal neurotoxins, microbial neurotoxins, bio-neurotoxins, and chemical neurotoxins, are mentioned earlier in this chapter. As for atypical neurotoxins, such as 3,5-DMA mentioned in the previous section, their effects are not immediate and not apparent, so their damage to the nervous system is not easy to detect. Instead, once it is detected, the damage is already severe and irreversible, such as neurodegeneration or developmental disorders [120, 121]. Some of these atypical neurotoxins are hidden in the surrounding environment, and some are obvious but neglected because of slow action, such as air pollution. Because there is no immediate risk, it is often ignored in clinical or in research. Therefore, how to find an efficient antidote or effective daily health care methods against this neurotoxin, especially in current aging society, become much more critical.
The authors have declared that no competing interests exist.
Ove Odredbe i uvjeti ističu pravila i regulacije u svezi korištenja IntechOpenove stranice www.intechopen.com i svih poddomena u vlasništvu IntechOpena, tvrtke sa sjedištem u 5 Princes Gate Court, London, SW7 2QJ, Ujedinjeno Kraljevstvo.
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\\n\\nSljedeća terminologija odnosi se na Odredbe i uvjete, te na sve naše ugovore:
\\n\\nKlijent, stranka, vi, vaš odnosi se na vas, osobu koja pristupa ovoj stranici i prihvaća IntechOpenove Odredbe i uvjete;
\\n\\nKompanija, tvrtka, mi, naše odnosi se na tvrtku IntechOpen;
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\\n\\nMi koristimo kolačiće. Korištenjem IntechOpenove stranice slažete se s korištenjem kolačića u skladu s IntechOpenovom Politikom privatnosti. Većina modernih, interaktivnih stranica koristi kolačiće kako bi omogućila ponovno pronalaženje korisničkih detalja kod svakog posjeta. Na našoj stranici kolačići se uglavnom koriste kako bi omogućili funkcionalnost i olakšali posjetiteljima korištenje stranice.
\\n\\nIntechOpen ili njegovi suradnici niti u jednom slučaju neće biti odgovorni za štete (štete uključuju gubitak podataka ili profita, druge poslovne prekide, te sve ostale štete) koje nastanu zbog korištenja materijala na IntechOpenovoj stranici ili nemogućnosti da se iste koriste, čak i ako je IntechOpen ili njegov predstavnik o takvoj šteti obaviješten pismenim ili usmenim putem. Neke jurisdikcije ne dozvoljavaju ograničenja garancija ili ograničenja obveza za posljedične ili slučajne štete pa se u tom slučaju ova ograničenja možda ne odnose na vas.
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\\n\\nIntechOpen može ove Odredbe izmijeniti u bilo koje vrijeme i bez prethodne obavijesti. Koristeći ovu stranicu vi se slažete s trenutnim Odredbama i uvjetima koje su na snazi.
\\n\\nOve Odredbe i uvjeti su sastavljeni u skladu s odredbama prava Ujedinjenog Kraljevstva, a za sve sporove nadležan je sud u Londonu, Ujedinjeno Kraljevstvo.
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\n\nSljedeća terminologija odnosi se na Odredbe i uvjete, te na sve naše ugovore:
\n\nKlijent, stranka, vi, vaš odnosi se na vas, osobu koja pristupa ovoj stranici i prihvaća IntechOpenove Odredbe i uvjete;
\n\nKompanija, tvrtka, mi, naše odnosi se na tvrtku IntechOpen;
\n\nStranke, strane odnosi se na klijenta i na nas, ili samo na klijenta ili nas.
\n\nSve odredbe koje se odnose na ponudu, prihvat ili razmatranje plaćanja, a za koja mi pružamo asistenciju klijentu, bilo na ugovoreni ili fiksni način, a s ciljem da se ostvare potrebe i želje klijenta u svezi s našim uslugama, su podložne zakonskim odredbama Ujedinjenog Kraljevstva.
\n\nOsim ako nije suprotno navedeno, IntechOpen i/ili svi davatelji licence vlasnici su intelektualnog vlasništva nad svim materijalima na www.intechopen.com. Sva prava intelektualnog vlasništva su pridržana. Stranice sa www.intechopen.com možete gledati, preuzimati, dijeliti, dijeliti poveznice i printati za osobnu uporabu, a temeljem pravila sadržanih u ovim Odredbama i uvjetima.
\n\nMi koristimo kolačiće. Korištenjem IntechOpenove stranice slažete se s korištenjem kolačića u skladu s IntechOpenovom Politikom privatnosti. Većina modernih, interaktivnih stranica koristi kolačiće kako bi omogućila ponovno pronalaženje korisničkih detalja kod svakog posjeta. Na našoj stranici kolačići se uglavnom koriste kako bi omogućili funkcionalnost i olakšali posjetiteljima korištenje stranice.
\n\nIntechOpen ili njegovi suradnici niti u jednom slučaju neće biti odgovorni za štete (štete uključuju gubitak podataka ili profita, druge poslovne prekide, te sve ostale štete) koje nastanu zbog korištenja materijala na IntechOpenovoj stranici ili nemogućnosti da se iste koriste, čak i ako je IntechOpen ili njegov predstavnik o takvoj šteti obaviješten pismenim ili usmenim putem. Neke jurisdikcije ne dozvoljavaju ograničenja garancija ili ograničenja obveza za posljedične ili slučajne štete pa se u tom slučaju ova ograničenja možda ne odnose na vas.
\n\nMaterijali koji se pojavljuju na IntechOpenovoj stranici mogu sadržavati manje greške, tipfelere ili fotografske greške. IntechOpen može napraviti promjene na bilo kojem materijalu koji se nalazi na stranici u bilo koje vrijeme.
\n\nIntechOpen nije formalno povezan niti s jednom vanjskom stranicom čije poveznice vode na www.intechopen.com, osim ako to nije izravno navedeno. Iz tog razloga IntechOpen nije odgovoran za sadržaj koji se pojavljuje na takvim stranicama. Poveznica na IntechOpenovu stranicu ne implicira povezanost sa IntechOpenom. Korištenje takvih poveznica isključiva je odgovornost korisnika.
\n\nZadržavamo pravo vlasništva nad cjelokupnom stranicom www.intechopen.com i nad svim materijalom na toj stranici. Koristeći se našim uslugama, slažete se da maknete sve poveznice na našu stranicu odmah nakon što to od vas zatražimo. Također, zadržavamo pravo da ove Odredbe i uvjete, i politiku o poveznicama izmjenimo u bilo koje vrijeme. Koristeći se poveznicama na naše stranice slažete se s ovim Odredbama i uvjetima.
\n\nAko smatrate da je bilo koja poveznica na našoj stranici sumnjiva iz bilo kojeg razloga, molimo vas da nas kontaktirate. U tom slučaju razmotrit ćemo micanje poveznice s naše stranice, iako nismo obvezni to napraviti.
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\n\nIntechOpen može ove Odredbe izmijeniti u bilo koje vrijeme i bez prethodne obavijesti. Koristeći ovu stranicu vi se slažete s trenutnim Odredbama i uvjetima koje su na snazi.
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I am also a member of the team in charge for the supervision of Ph.D. students in the fields of development of silicon based planar waveguide sensor devices, study of inelastic electron tunnelling in planar tunnelling nanostructures for sensing applications and development of organotellurium(IV) compounds for semiconductor applications. I am a specialist in data analysis techniques and nanosurface structure. 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