\\n\\n
Dr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\\n\\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\\n\\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\\n\\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\\n\\nThank you all for being part of the journey. 5,000 times thank you!
\\n\\nNow with 5,000 titles available Open Access, which one will you read next?
\\n\\nRead, share and download for free: https://www.intechopen.com/books
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Preparation of Space Experiments edited by international leading expert Dr. Vladimir Pletser, Director of Space Training Operations at Blue Abyss is the 5,000th Open Access book published by IntechOpen and our milestone publication!
\n\n"This book presents some of the current trends in space microgravity research. The eleven chapters introduce various facets of space research in physical sciences, human physiology and technology developed using the microgravity environment not only to improve our fundamental understanding in these domains but also to adapt this new knowledge for application on earth." says the editor. Listen what else Dr. Pletser has to say...
\n\n\n\nDr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\n\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\n\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\n\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\n\nThank you all for being part of the journey. 5,000 times thank you!
\n\nNow with 5,000 titles available Open Access, which one will you read next?
\n\nRead, share and download for free: https://www.intechopen.com/books
\n\n\n\n
\n'}],latestNews:[{slug:"intechopen-authors-included-in-the-highly-cited-researchers-list-for-2020-20210121",title:"IntechOpen Authors Included in the Highly Cited Researchers List for 2020"},{slug:"intechopen-maintains-position-as-the-world-s-largest-oa-book-publisher-20201218",title:"IntechOpen Maintains Position as the World’s Largest OA Book Publisher"},{slug:"all-intechopen-books-available-on-perlego-20201215",title:"All IntechOpen Books Available on Perlego"},{slug:"oiv-awards-recognizes-intechopen-s-editors-20201127",title:"OIV Awards Recognizes IntechOpen's Editors"},{slug:"intechopen-joins-crossref-s-initiative-for-open-abstracts-i4oa-to-boost-the-discovery-of-research-20201005",title:"IntechOpen joins Crossref's Initiative for Open Abstracts (I4OA) to Boost the Discovery of Research"},{slug:"intechopen-hits-milestone-5-000-open-access-books-published-20200908",title:"IntechOpen hits milestone: 5,000 Open Access books published!"},{slug:"intechopen-books-hosted-on-the-mathworks-book-program-20200819",title:"IntechOpen Books Hosted on the MathWorks Book Program"},{slug:"intechopen-s-chapter-awarded-the-guenther-von-pannewitz-preis-2020-20200715",title:"IntechOpen's Chapter Awarded the Günther-von-Pannewitz-Preis 2020"}]},book:{item:{type:"book",id:"2555",leadTitle:null,fullTitle:"Lipoproteins - Role in Health and Diseases",title:"Lipoproteins",subtitle:"Role in Health and Diseases",reviewType:"peer-reviewed",abstract:"By typing into databases such as Medline or PubMed the word “lipoprotein” one gets more than 100.000 hits that highlight the common interest in this topic. It is actually impossible to cover all aspects of lipoprotein structure, function, metabolism and pathophysiology in one issue like the present volume, but attempts have been made to concentrate on topics that are in focus of current lipoprotein research. These topics have been divided into 10 sections. 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After his studies of Chemistry and Physics he got a PhD in Biochemistry and spent two years at OMRF, USA with Pierre Alaupovic and half a year as guest professor at the University of Frankfurt, Germany. Further details of his academic carrier are found in http://www.medunigraz.at/mbbc/cms.php?pageName=GMK",institutionString:null,position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"2",institution:{name:"Medical University of Graz",institutionURL:null,country:{name:"Austria"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:{id:"136970",title:"Prof.",name:"Sasa",middleName:null,surname:"Frank",slug:"sasa-frank",fullName:"Sasa Frank",profilePictureURL:"https://mts.intechopen.com/storage/users/136970/images/4959_n.jpg",biography:"Saša Frank is Associate Professor at the Institute of Molecular Biology\nand Biochemistry, Medical University of Graz, Austria. 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\r\n\tEvolutionary and developmental biology is focused on the investigation and the elucidation of the molecular mechanisms underlying embryonic development from one generation to the other. It started quite early with Darwin and Mendel, but soon it became apparent that it is a complicated network of interacting genes after stimuli from molecules, our microbiome, other organisms or species and the environment that we live in. However, in recent years the accumulation of big data in the fields of genomics, medical records and other relevant datasets has rendered the study of evo-devo impossible without the aid from advanced and highly sophisticated computational models, which aim to mine and fuse information from diverse disciplines in the realm of evolution and developmental biology.
\r\n\tThis book aims to address the new developments in the rapidly evolving field of evo-devo in the post genomics era. All recent biological and medical breakthroughs in the evo-devo field are welcomed. Finally, review articles encompassing recent advances, development current and future trends are also more than welcomed.
Surgical rhinoplasty remains one of the most popular cosmetic procedures performed on the face. Statistics from the American Academy of Facial Plastic Surgery and the American Society of Aesthetic Plastic Surgery put the number of rhinoplasties performed in the United States in 2012 at around 190,000 [1,2]. This number is relatively unchanged from what it was in 1997, prior to the FDA approval of cosmetic botulinum toxin and hyaluronic acid fillers. As with any surgical procedure, patients undergoing rhinoplasty are subject to significant risks, recovery time and expense. Until recently, patients who wanted to avoid surgery have never had a viable alternative procedure that could accomplish the cosmetic goals of rhinoplasty noninvasively. With the advent of long-lasting injectable fillers, however, physicians and patients have embraced a nonsurgical surrogate.
I first performed primary Nonsurgical Rhinoplasty (NSR) using calcium hydroxyapatite injectable filler in 2003, prior to the publication or report of such a procedure being done in the United States. The novelty of the procedure caught the attention of the media, and I performed it on several prominent national news programs and shows. This sparked the interest of the public and doctors around the country began to offer NSR to their patients. Several small studies have documented the safety and efficacy of the procedure since then [3,4,5]. Due to the simplicity and efficacy of NSR, it has steadily grown in popularity over the last decade. Since 2003, I have had the privilege to perform over 2500 of these procedures in my clinic, using a variety of injectable fillers, with great success.
The idea of injecting substances under the nasal skin to improve cosmesis is not new. Corning and Gersuny first described injecting liquid paraffin into the nose to correct saddle nose deformity at the beginning of the nineteenth century. The practice was quickly embraced but just as quickly abandoned because of the severe long-term adverse effects of paraffin [6]. In 1919, Bruning tried to correct postoperative cosmetic nasal imperfections with fat injection [7]. The technique was ultimately not very well accepted because the fat grafts showed poor survival duration [8]. In 1986, Webster presented a 20-year retrospective study of microdroplet silicone injection into the nasal bridge to correct postsurgical nasal defects [9]. The study reported mostly positive results, but many physicians, aware of horrific reports of complications from the 1960s and 1970s, have remained wary of silicone injections.
The first modern study of NSR was published by Han et al. in 2006 [10]. It was an 11-patient pilot study, looking at the safety and efficacy of dorsal augmentation NSR using hyaluronic acid (Restylane, Medicis, Scottsdale, Ariz.) mixed with autologous fibroblasts. Hyaluronic acid (HA) is a glycosaminoglycan that is a major component of connective, dermal, and neural tissue in most mammals. Cross-linked HA has been widely used as an injectable filler since it was FDA approved in 2003. Because of its biocompatibility, no allergy testing is required. The HA in Restylane is produced by bacterial fermentation. The authors injected an average of 0.8 cc into the nasal dorsum of their patients, overcorrecting by 20%. Of the 6 patients that Han et.al were able to get follow-up data on, there were no adverse events and the augmentation effect was still persisting at 12 months. Later that year, Beer published a case report where he successfully corrected a dorsal cosmetic defect with Restylane [11].
Also that year, Laryngoscope published a report by Nyte where he successfully injected calcium hydroxyapatite (CaHA, Radiesse, Merz, San Mateo, CA) to correct collapse of the internal nasal valve in 23 patients [12]. Radiesse is a suspension of 30% of CaHA microspheres, 25–45µm in size, in a mix of glycerin, carboxymethyl-cellulose, and water. It is fully biocompatible because CaHA is identical to the mineral portion of human bone and teeth [13]. Radiesse was first approved as a radiologic marker and for use in vocal fold augmentation. FDA approval for cosmetic use came in 2006. Nyte’s technique of injecting through the inside of the nose seemed to show some cosmetic correction along with the functional improvement. This is, however, the only study that advocated intranasal injection.
Several papers emerged in the next year, including two small studies of Radiesse for cosmetic improvement of the nose. In the paper by Stupak et al., the authors reported on a 13-patient prospective single-arm trial with blinded evaluators of before and after pictures [5]. They injected post-rhinoplasty patients seeking minor contour improvement. Areas injected included dorsum, supratip, nasal sidewall, and ala. Mean amount injected was 0.18cc. They did not inject the nasal tip due to concern that Radiesse would diffuse after injection and the tip would lose definition. There were no adverse events and patient satisfaction and evaluator ratings were good, but follow-up was only 2 months. Dayan et al. described their experience with Radiesse NSR in 8 patients over the span of 2 years [14]. The volume they used ranged from 0.3cc to 1.6cc and the corrections were mostly to the dorsum, radix, and supratip. Duration of effect was estimated to be around 1 year. They encountered no complications. At the end of their paper they made a point of mentioning that Dayan had performed revision rhinoplasty on a patient who had received Radiesse 14 months earlier without complication.
The other paper that year was by De LaCerda et al. from Brazil [15]. They presented a 2-patient experience with NSR using small amounts of porcine collagen on one patient (0.35 cc) and hyaluronic acid (Voluma, Allergan, Irvine, Calif.) on the other (0.20 cc); follow-up was 4 months and 1 year, respectively. Areas injected included the nasofrontal angle, nasolabial angle, the dorsum, and the tip.
In 2009, Humphrey and Dayan published a paper describing their preferences and techniques of NSR in 22 patients with HA and an unspecified number with CaHA [16]. They advocated using CaHA over HA because the latter absorbed water unpredictably. They considered injecting significant amounts of HA into virgin noses dangerous due to the risk of vascular compromise from the hydrophilic swelling of the product. They recommended injecting filler subdermally and avoiding the tip and supratip area. The only complications that they observed from the technique came after injecting HA into the tip area of two post-rhinoplasty patients, one by Dayan and one by an outside injector. These were both cases of ischemia. Dayan’s patient was a mild case of Raynaud’s-like phenomenon on the nasal tip and had no sequelae due to treatment with Hyaluronidase. The outside injector’s case was a more serious case of ischemia that resolved with Nitropaste and Hyaluronidase. Although necrosis was avoided, they report that the patient developed aesthetically displeasing skin changes 1 year afterward. Notably, they again write that Dayan has had no trouble performing rhinoplasty on post-CaHA NSR patients. This is interesting because it contradicted numerous anecdotal accounts by other rhinoplasty surgeons at meetings and on the Internet that injectable fillers were causing catastrophic scarring and damage to the nasal tissues [17].
In 2010, we published our 4-year experience with CaHA NSR, comprising 385 patients [18]. We injected virgin noses as well as patients who had undergone rhinoplasty surgery in the past. We used volumes ranging from 0.3 cc to 0.5 cc of CaHA. We showed the procedure to be extremely safe. Two serious complications were encountered in patients who had undergone multiple revision rhinoplasties. They consisted of ischemia that progressed to small areas of tissue necrosis at the tip and ala despite treatment with Nitropaste and oral steroids. The rate of minor complications like cellulitis, prolonged swelling, or prolonged bruising was very low. Interestingly, a history of previous rhinoplasty surgery did not increase the risk of minor complications. The only exception was that post-rhinoplasty patients did have an increased incidence of prolonged erythema. We documented successful injection of all areas of the nose, including the tip, ala, dorsum, sidewall, and radix. We were surprised that the cosmetic effect of CaHA did not, on average, last as long as we expected. A significant number of patients showed evidence of resorption of the material as early as six months after injection.
In 2012, Kim and Ahn published a paper on a standardized NSR technique for Asian patients using mostly CaHA (except for tip injection, where they used HA) [19]. They reported their experience with 87 patients. Unlike most authors, they used 2% Xyocaine for anesthesia, injecting the infratrochlear and external nasal nerves as well as placing boluses at the nasal tip and the columnella-labial angle. Their technique consists of three steps. First, the columnella and columnella-labial angle is augmented with an injection from the tip down to the nasal spine. Second, the dorsum is augmented with a single injection, advancing the needle from the tip to the radix and injecting upon withdrawal, as in the first step. The third step involves shaping the tip with small, superficial bolus injections of HA. Four minor complications are reported, none of them ischemic. They end the paper with a series of sensible guidelines to avoid complications and achieve optimal cosmetic results.
In 2013, Kurkjian and Rohrich published their recommendations on technique for NSR [20]. The extent of their experience with the technique is not stated. They advocate low-pressure injection with HA fillers only, using the smallest possible needles. According to them, Sculptra and Radiesse should be avoided due to their irreversibility and the danger of long-term palpability under relatively thin nasal skin. For patients desiring permanent nonsurgical correction, permanent fillers should be avoided and fat should be used instead. They recommend using Restylane in all areas of the nose because it is relatively less hydrophilic. The exception is the tip, where Juvederm is also recommended, taking advantage of postinjection hydrophilic swelling for patients who want to increase tip fullness. In their experience, HA has much higher longevity in the nose, lasting up to 2–3 years in some areas (I have not seen evidence of this kind of duration in my HA NSR patients). They consider filler injection to be useful in the correction of post-rhinoplasty contour irregularities.
This year I published a prospective, blinded study on the safety and efficacy of injecting methyl methacrylate (Artefill, Suneva, Santa Barbara, CA) to correct nasal contour deficiencies [21]. Artefill is a third-generation methyl methacrylate filler in a collagen carrier that was FDA approved in 2006 for nasolabial fold correction. The filler is 20% methyl methacrylate and 80% bovine collagen. Allergy testing for the carrier is commonly done 2 weeks prior to injection. The particles are smooth and 30–50 microns in size. I have been using Artefill off-label for NSR since 2006 with no complications in about 750 patients. In my experience, 3–5 injection sessions are necessary because the body quickly absorbs the collagen carrier and the methyl methacrylate stimulates fibroblast proliferation to a variable degree. For the study, I injected the product over 3 sessions, spaced 1 month apart. Results were evaluated by me, an independent, blinded MD evaluator, and the patient. With 1 year follow-up on 19 patients, we all observed excellent cosmetic effect and no complications of any kind. An example of the results we obtained is illustrated in Fig. 3.
NSR corrects mild or moderate cosmetic nasal irregularities. I perform this procedure to achieve the following cosmetic goals:
Raise and better define an underdeveloped nasal dorsum. This is most popular among my Asian patients.
Raise and better define a ptotic tip, as in Fig. 1.
Camouflage a dorsal bump. The dorsum is leveled by injecting filler above and below the bump, as in Fig. 2.
Correct asymmetry of the tip or dorsum by subtly augmenting the weaker side.
Correct post-rhinoplasty contour defects. Most commonly, these present as saddle nose deformity or other type of dorsal cartilage collapse, polly beak deformity, dorsal asymmetry due to asymmetric scarring, asymmetry of the tip due to postsurgical scarring or cartilage over-resection and alar foreshortening.
CaHA NSR to lift a ptotic nasal tip and augment the nasal radix. The “Before” pictures are on top, “After” on the bottom. The net result is a straighter nose on profile that appears smaller because it blends better into the rest of the face.
Injectors around the world have used a variety of materials to achieve the aesthetic recontouring of NSR. Currently, hyaluronic acid (HA) is the most popular filler material due to its reversibility (via injection of Hyaluronidase). There are a number of reports and editorials on technique in the literature. Most of these have been summarized above.
HA NSR to camouflage a dorsal hump. The patient wanted to use a minimum of product to make her profile straighter. She believed a slight curve would make her nose look more natural.
I consider HA, in its most common formulations of Juvederm and Restylane, to be the best material for the beginner or the occasional injector of the nose. The ability to dissolve HA is a critical safety feature for injectors who are not yet experts in the technique. Juvederm and Restylane are effective for basic NSR goals, such as augmenting the bridge and camouflaging minor dorsal bumps. These fillers struggle to perform, however, in the more advanced applications of this technique. Juvederm and Restylane are relatively soft materials. They can only provide moderate augmentation of the dorsum, they cannot lift a drooping tip very much and they are poor at sculpting defining points of the tip and the sidewall of the nose. In my experience, these HA fillers last an average of 6–8 months.
Perlane is a formulation of HA that is an improvement over Juvederm and Restylane for NSR. Its increased density permits the advanced injector to sculpt more effectively. It has less of a tendency to spread, so better definition can be achieved. Perlane also lasts somewhat longer in the nose than Juvederm and Perlane – about 8 months in my experience. Prior to the FDA approval of Voluma, I used Perlane for patients who wanted the added safety of a reversible filler.
I have used calcium hydroxyapatite (CaHA – Radiesse, MERZ Aesthetics Inc., San Mateo, CA) in most of my NSR procedures. Radiesse received FDA approval in 2006 for correcting moderate to severe wrinkles and HIV-related facial volume changes. Prior to that it was approved for vocal cord augmentation and I was using it off-label in the nose. As a non-HA filler, CaHA is not reversible. Hypersensitivity reactions to CaHA are extremely uncommon and reports of other adverse events do not differ significantly from reports for HA fillers. Side effects noted of CaHA include:
Nodules (generally associated with lip augmentation or injection in areas with little subdermal space, e.g., infraorbital area) (Smith, 2007) (Tzikas, 2008)
Granulomas (Lee MJ, 2008)
Skin necrosis (Dayan SH and Arkins JP, 2011)
Redness, erythema, and swelling (Siclovan, 2009)
The advantages of CaHA are its relative persistence of effect (average of 9–10 months in my experience, but quite variable between 6 and 12 months) and its high density. This last quality allowed me to effectively sculpt noses to my patients’ satisfaction. It is possible to significantly elevate a droopy tip without excessive rounding. In fact, CaHA makes it possible to precisely create aesthetically pleasing tip defining points in patients with rounded and poorly defined tips. CaHA also makes it possible to significantly raise and add real definition to an underdeveloped dorsum – a quality that my Asian patients particularly appreciate.
With the FDA approval of Voluma in October of 2013, injectors gained a valuable new tool that seems to confer longer duration of effect than any other filler. Voluma is more cross-linked than other HA fillers and has a higher percentage of low molecular weight hyaluronic acid, making it exceptionally smooth, viscous, and cohesive. Increased cross-linking makes the filler more resistant to enzymatic degradation. Under study conditions, duration of effect was up to 2 years [22], but anecdotal reports from the investigators indicate the filler to be even longer lasting than that. Because of the duration of effect and its reversibility, I have been using Voluma for the majority of my NSR cases over the last 5 months. It performs well for the most part, but I find that in patients that need extensive elevation of their tip or dorsum, Radiesse is still the only filler thick enough to provide the desired lift. In these patients, my NSR combines Voluma and Radiesse.
Artefill NSR to refine and straighten the dorsum and tip from both the profile and straight on views. This patient wanted to have a thinner appearing nose from the frontal view and a straighter and more refined looking dorsum and tip from the profile.
I perform this procedure with the patient sitting up straight in the chair, as in Fig. 5. After taking standardized photographs, I use a compounded triple anesthetic cream for 15–20 min prior to the procedure (the materials I use are pictured in Fig. 4). My assistant will remove the cream and the patient will ice the area to be injected. During the injection, my assistant taps the patient’s contralateral shoulder to distract their attention from the injection. I have found that alcohol works fine to clean the area and prevent infections. I prefer a thin-walled 29 gage half-inch needle when using any of the fillers – Voluma, Radiesse, or Artefill. If that is not available, I use a thin-walled 28 gage one-inch needle. I perform injections for the most part as shallow linear threads, placing small amounts of filler as I withdraw the needle. I will place filler into the area of the radix, dorsum, sidewall, tip, columnella, and ala as needed to correct each individual irregularity. I will then massage and mold the filler to blend into the desired contour. The patient goes home with instructions to avoid alcohol and strenuous exercise that day, as well as heavy sunglasses for two weeks. I agree with Kim and Ahn’s observation that the volume effect of most fillers decreases by about 25% within the first month or two [19]. For this reason, my procedure includes a complementary follow-up visit 3 weeks after the initial injection, where I can touch-up the results.
Equipment necessary for NSR. Radiesse, Voluma, and Perlane are all displayed with the first two as presplit half syringes (using a Luer-Lock connector and a 1cc syringe). I try to use the smallest possible needle gage – 27 or 28 G thin-walled needles are my preference. The photograph displays the four main methods of pain control in my injectables practice. Topical numbing cream is a compounded 23% Lidocaine, 7% Tetracaine mixture. Ice and a stress ball are important, but most important is my assistant’s hand, which distracts the patient by tapping on the shoulder opposite to where I am standing.
In my experience, there are several types of patients that seek NSR. Most commonly, it is the younger patient who cannot afford the time or resources required for postsurgical recovery. They hear about the procedure through friends and the Internet. They mostly present with mild or moderate cosmetic irregularities. Having studied the before and after pictures and read about the procedure, they understand that it is not a technique that can physically reduce the size of the nose, so I rarely see patients with severely large noses.
Correct patient position for NSR. She is relaxed and sitting comfortably with her head resting on the back of the chair. Movement is minimized. There is no tissue redistribution, as there would be if the patient was lying flat, so cosmetic results are optimally accurate.
About a third of my NSR patients are Asian, as in Fig. 6. These patients commonly desire an increase in the height and definition of their dorsum and radix, as well as improvement in the definition and projection of their nasal tip. These patients are usually young and cannot afford surgery. They are also often wary of unnatural surgical results, describing people with visible or overly large dorsal grafts that they have seen in the Asian community. Like many patients who opt for this procedure, they want to see the cosmetic change they desire, but only if their nose retains a natural appearance and no one can tell that they have had an aesthetic procedure.
CaHA NSR to augment and better define a nasal bridge and tip in an Asian patient.
Both of these kinds of patients usually choose the temporary procedure using a CaHA filler like Radiesse or an HA filler like Perlane or Voluma. They are aware that I commonly use Artefill for long-lasting to permanent results but want to try out the effect before they commit. Once the effect fades, some of these patients decide to continue with the temporary filler but many switch over to Artefill.
Artefill NSR to correct postsurgical contour irregularities and asymmetry of the dorsum and to lift the nasal tip. Lower series are 1 year after the last of three sessions of Artefill injections.
Another common category of patients presenting for this procedure are the ones who have desired cosmetic improvement in their nose for a long time, but have been afraid of surgical and anesthesia risks. These patients present with a variety of aesthetic complaints. Most are appropriate for the NSR technique, but some require reduction and have to be turned away.
An important subset of the above patient group is those who consider their aesthetic complaint to be too minor to undergo surgical correction. They are bothered by their small bump or mild asymmetry but feel that surgery exacts too high of a price (both financial and temporal). These patients are mostly ideal candidates for NSR. A small amount of contour correction to restore symmetry or camouflage a small bump makes them very happy.
About a quarter of my patients have already had one or more surgical rhinoplasties. These patients present with a variety of aesthetic complaints, but all of them are disappointed in the aesthetic result of their surgery (or surgeries) and desire an effective alternative. Some opt for temporary fillers, but many choose methyl methacrylate so that they can “get it over with.” Figure 7 illustrates one of those patients. Anyone who performs NSR must be aware that these patients present with technically challenging problems. Postsurgical scarring stiffens the skin and limits the lift that can be achieved with filler injection. Injectors need to take care not to over-promise these patients. Their results are going to be, for the most part, relatively subtle. Most importantly, postsurgical skin has a more tenuous blood supply, especially around the tip and ala. The risk of ischemia and necrosis are significantly increased in these patients. Only the most experienced injectors should be treating them, since the complications of necrosis can be catastrophic.
Complications of NSR are relatively rare. This is not surprising, since the overall major complication rate for filler injections has been estimated to be less than one hundredth of one percent [4]. As described above, published studies are meager and mostly small, but they report few serious adverse events. Bruising, transient erythema, and short-term swelling represents most of the issues documented. The exceptions are the disturbing case reports that detail blindness and major necrosis of nasal tissue [5,6,7]. Since the doctors treating the complications and not those who injected the patients write most of these reports, conclusions about needle type, injection technique, and even material used are often difficult. Of the reports published by the actual injectors, we know that sudden pain, blanching, duskiness, and ecchymosis in the area being injected are all danger signals for ischemia and necrosis. Compromise of the blood supply to the skin in cases of filler injection can be caused by either intravascular embolism or small vessel compression by the filler.
Opthalmalgia and visual loss within minutes are signs of retinal artery embolism. Other signs of ophthalmic vasculature embolism include immediate diaphoresis, nausea, headache, opthalmoplegia, and ptosis [4].
In my experience, these complications can best be avoided by understanding the anatomy, always practicing safe injection technique, and having well-prepared protocols ready to launch at any sign of danger. Our traditional understanding of the vascular anatomy of the external nose is illustrated in Fig 8.
External arterial anatomy of the nose. Vessels originating from the External Carotid are in red, whereas those in black come from the Internal Carotid via the Opthalmic Artery.
A good injector is especially careful when injecting the radix and nasal sidewall, to avoid the dorsal nasal and angular branches, respectively. Based on Fig. 8, keeping one’s injection points in the midline seems to ensure safety. This is surely true to some degree, but a recent paper by Saban et al. proposes that the external nasal vasculature is more interconnected than we think [23]. Using cadaver dissection and ultrasonography study of live subjects, the authors conclude that anastomoses between the internal and external carotid vascular systems are plentiful in the external nose. This plexus of vessels is located in the SMAS layer. Safe injection technique should therefore focus on keeping the needle deep instead of trying to avoid specific vessels.
Some clinicians have recommended aspiration prior to injection of filler in order to avoid vascular embolism. In my experience, this is a cumbersome practice with questionable benefits. Most fillers are thick gels. Building up enough negative pressure for aspiration of blood takes a lot of hand force and would only work for Voluma due to its smooth viscoelastic properties. Even in experienced hands, by the time one aspirates and then injects, the needle is no longer in the same place, rendering the test useless.
Rather than aspirating, I think that following accepted best practice standards for injecting filler is a more reliable way of preventing complications [8,9]:
Needles should be as small as possible so that filler flow rate is low.
Fillers should always be injected slowly and under low pressure, especially in the nasal area. The blindness complications in the literature occurred because a filler embolus was injected with enough force to overcome systolic blood pressure and travel up into the ophthalmic vasculature. Gentle injection technique should prevent this complication.
The needle should be advanced through the skin slowly and filler should only be flowing when the needle is moving out of the skin. This way, even if the tip of the needle is inside the lumen of a vessel at some point, only a tiny amount of filler will enter the vessel, as the needle will be out in the next moment.
Small volumes of filler should be introduced with each injection.
We always ice prior to injection. Ice decreases pain and shrinks blood vessels, making them less likely to be punctured.
Some authors advocate blunt tipped cannulas for all filler injection as a way to reduce complications and discomfort [10,11]. I think this is not a good idea in the nose. The weakness of cannulas is precision. It is much easier to know the exact location of a needle tip than the tip of a cannula that bends easily as it is advanced through the tissue. The aesthetics of the nose are particularly sensitive to the smallest contour asymmetries. One millimeter of fullness difference between two sides of the tip is clearly noticeable. One-millimeter deviation from the midline, when augmenting a dorsum, makes the nose appear off-center.
The literature contains reports of severe complications from every type of filler available, and I have successfully used a variety of materials in the nose. I do not believe that there is a material contraindicated for nasal injection. It is the skill and knowledge of the injector that is paramount to the success of the procedure.
While not really a complication, vaso-vagal episodes can be disturbing to the novice injector. Diaphoresis, sudden pallor, a sensation of nausea are all warning signs of an impending episode. It is more likely to happen if the patient has not eaten much prior to their appointment. In the setting of cosmetic injections, patients become vaso-vagal primarily because they are holding their breath in anticipation of pain. The frequency of these episodes dropped dramatically once we began to routinely remind our patients to breathe. Distraction shoulder tapping also helps them tolerate the injections without excessive anxiety. If the patient does become vaso-vagal, our routine is to immediately place them in reverse Trendelenberg position, increasing blood flow to the brain. We place an ice bag behind their neck and give them something sweet to drink like orange juice or Coke. We monitor their pulse and blood pressure manually. Most patients will recover fully within a few minutes. We have not yet had to use smelling salts.
Complication management starts with the preventative measures outlined above. If, however, the clinician suspects that an ischemic event is unfolding, there are immediate steps that he or she should be ready to take. First, injection should stop immediately. The area should be massaged vigorously in an effort to restore blood flow and Hyaluronidase should be injected into the area. A dose of 50–80 units should be sufficient. Even if a non-hyaluronic acid filler has been used, Hyaluronidase is useful because it dissolves some native hyaluronic acid and decreases interstitial pressure, easing blood flow. Topical 2% Nitroglycerin paste (Nitro-bid. Savage Labs, Melville NY) should be in the room and readily available to anyone performing this procedure. It is a great vasodilator that acts very quickly. In situations of potential ischemia, a small amount should be applied to the area in question and it should stay on for at least 15 min. The patient should take aspirin 325 mg immediately. If the skin becomes pink again and remains so after a period of observation of 15 min or so, I would feel comfortable sending the patient home on aspirin every 4 h for a day, warm compresses, and periodic massage of the area. If the skin becomes dusky, I would reapply the Nitropaste for another 30 min and consider reinjecting the area with Hyaluronidase. I would observe the patient in the office for the next hour or so, applying warm compresses and massaging the area. At this point, the patient should begin oral steroid therapy and oral antibiotics. I would give them a Medrol dose pack and make sure that they take the first dose right away. At this point, the patient should be seen in the office on a daily basis to monitor the progression of tissue damage. If damage continues to unfold, the injector should consider hyperbaric oxygen therapy and referral to a plastic surgeon. My in-room safety kit, a.k.a. injectables “crash cart” is illustrated in Fig. 9.
My in-room safety kit. Pictured are Hyaluronidase, Nitopaste, Aspirin, Kenalog, Solu-medrol, and smelling salts. These are the medications important to have on hand in every room.
When performed safely and correctly by an experienced and well-trained injector, NSR is a procedure that yields excellent patient satisfaction. It is not a replacement to rhinoplasty for all patients and I continue to refer those that are not candidates (mostly patients who need significant reduction) to my surgical colleagues. This procedure does, however, provide a valuable alternative to traditional surgery. It increases the pool of people wanting cosmetic correction of the nose, bringing in a significant population of patients who would never do surgery. In fact, recalling the statistics from AAFPRS and ASAPS, over the last 10 years that this procedure has become popular, it has not cut into the number of patients receiving surgical rhinoplasty [1,2]. For the patients who are candidates, it saves them expense, risk, and downtime. Finally, the precision of filler injection means that this procedure is in some cases superior to surgery in accomplishing patients’ cosmetic goals. I have built a large practice upon this procedure and continue to receive many referrals from happy patients.
The author would like to thank Mimi Lam for her assistance with the layout and proofreading of this chapter.
Higher multicellular organisms have coexisted and co-evolved with resident microorganisms in a relatively harmonious relationship over millions of years, forming a complex organism called holobiont. These processes of co-evolution have been documented by several studies carried out in the organization and composition of host microorganisms (microbiome) in different species [1, 2, 3, 4]. The microbiome is currently considered a functional organ which is fundamental for the host organism, given that studies have shown that this organ is highly dynamic and adaptable, likewise plays an important role in physiological adaptation processes, metabolism, and development [1, 2, 4, 5, 6]. The study of the role of the microbiome for years was limited to those organisms that could be susceptible to culture, the use of techniques based on molecular information (AFLP and RFLP) and denaturing gradient gel electrophoresis (DGGE) could reveal the presence of some species not cultured, but the gel resolution was being the main limitation since a single band could contain more than one sequence [7]. The development of genomic tools such as the new generation sequencing platforms (NGS) has allowed a better resolution of the diversity present in a given sample through what is known as metagenomics [2, 7, 8, 9, 10]. These same sequencing platforms have allowed not only observing the diversity of a host or environment, but also the functional role of microorganisms as well as their possible interactions with physiological processes or biogeochemical cycles through metatranscriptomics [1, 2, 4, 9]. The understanding of the composition of microorganisms, functions, interactions, and other biological processes through NGS has been done through the development of different bioinformatics tools, which make use of large amounts of information and are able to compare them through different data bases [3, 9, 11]. The objective of this chapter is to provide information on the different existing bioinformatics tools that have been used in studies of co-evolution and symbiosis in different models.
\nThe first step of a NGS-based study involves the extraction of nucleic acids in sufficient quantity and quality to carry out the sequencing process in order to have an unbiased knowledge of the microbial diversity present in a sample [6, 12, 13]. The processing of DNA samples (environmental and host) can be performed by cell recovery by centrifugation gradients in differential media and the subsequent recovery of DNA by silica columns [6]. Another methodology used is the in situ lysis of the sample by the addition of enzymes (Proteinase K and lysozyme) with the subsequent separation of cell debris by centrifugation and recovery of DNA by solvent precipitation or by silica adsorption [14]. The main advantage of in situ lysis is that higher amounts of DNA are obtained when compared to cell recovery techniques; however, there is a risk of the presence of contaminants that may interfere with sequencing reactions [6].
\nIn the case of RNA, the main methodologies perform in situ lysis of the sample under RNase-free conditions using different guanidine solvents and salts to avoid the presence of ribonucleases [15]. The samples should be placed at −80°C either in dry ice or liquid nitrogen to avoid their degradation.
\nQuality control of nucleic acids can be carried out by visualization on agarose gels, by spectrophotometric means (Nanodrop) and in microfluidic chambers (Bioanalyzers). This last system has been widely used since it allows the visualization and simultaneous quantification of nucleic acids [8, 16, 17, 18]. In the case of RNA, these systems have developed a scale known as RNA Integrity Number (RIN) which, based on the proportion between the major and minor subunits of the rRNA assigns a minimum value that must be greater than 8.0.
\nOnce a sample with sufficient quality and quantity was sent to sequencing, a series of files with the “.fastq” extension are obtained, which contains the information of the sequence and the quality for each base. This format is used by different programs (FASTQC and PRINSEQ) to perform the quality control of the sequencing, showing basic statistics such as the total number of bases, read size, GC content, quality for each base in PHRED33 or PHRED64 scale, as well as the presence of overrepresented sequences [8, 19, 20, 21, 22, 23]. The files analyzed are introduced to different programs (Trimmomatic, TrimGalore, and CutAdapt) that trims the reads of the “.fastq” file, based on the quality for each nucleotide, eliminating sequences with a PHRED value below 20 and a minimum fragment size defined by the user [19, 20, 22, 24].
\nThese programs are able to eliminate segments of initiators and sequencing adapters, which must be provided in a separate file. The output files of these programs are archives in “.fastq” format, where the sequences that are common for all samples are placed in one file, and the unique sequences for each individual sample are placed in several files [19, 20].
\nIn recent years, the use of genomic approaches has revealed an unprecedented diversity and bacterial ubiquity in different types of samples (Figure 1), through the analysis of 16S ribosomal sequences [1, 2, 5, 6, 18, 19, 22, 25, 26, 27]. These techniques have allowed the molecular analysis of populations and how different biological processes have been established, controlled, and evolved [5, 28, 29].
\nGenomic and metagenomic techniques for the analysis in different samples.
The metagenomic composition analyzes have been carried out through the use of different programs (QIIME, QIIME2, and MOTHUR), that align the reads against a database of ribosomal genes (GreenGenes, SILVA, and RDP) and assign them operational taxonomic units (OTUs), using a distance of 3% and a confidence interval of 80% [29, 30, 31, 32, 33]. Once the OTUs have been assigned, the aforementioned programs allow the determination of diversity indices, richness, and main component analysis and perform the rarefaction of the samples [1, 2, 5, 19, 25, 29, 30, 31, 32, 34, 35].
\nOther taxonomic classifiers are based on alignment of short sequences previously edited, by single or paired ends (Kraken, Kraken2, OneCodex) comparing them with the databases available in each program. In the case of Kraken, it makes use of the Ref-Seq database where the reads are divided into fragments known as k-mers and are compared with sequenced genomes [34, 36]. The resulting files of these programs are provided in tabular format (tsv), which facilitates their export and processing in other types of programs such as Vegan or R, where studies of richness, diversity, and rarefaction can be carried out [12, 34, 35, 37, 38].
\nThe use of different taxonomic binning programs has been able to determine the presence of ubiquitous microbial phyla present in samples from arctic, temperate, and tropical environments such as: Proteobacteria, Actinobacteria, and Cyanobacteria, which are considered cosmopolite phyla. The main difference between each site is the proportion of each taxa, which reflects the conditions of each environment [6, 8, 9, 10, 39, 40, 41]. A similar behavior has been observed when studying the microbiome in different animal models where the phyla: Proteobacteria, Acitnobacteria, Firmicutes, and Bacteroidetes have been reported among those of greater relative abundance [4, 9, 39, 42, 43, 44, 45, 46]. This shows that microbial communities are highly dynamic where the physical-chemical factors of the site, health status, and nutrition shape the metagenome and can determine how reactive a microbial community is to environmental changes.
\nThe use of genomic tools has made possible to identify the core microbiome of different organisms, given that, despite living in different habitats, they share similar bacterial communities, which implies the existence of biological filters that shape the bacterium-host interactions, resulting in a stable relationship with the holobiont [2, 28, 45, 46, 47]. In the case of Apis mellifera, a global core microbiome formed by Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria has been identified, together with a high amount of lactic acid bacteria which have a beneficial activity in the health of the host organism due to their involvement in the immunomodulation of the intestinal microflora [25, 39, 48]. The presence of symbiont microorganisms within the intestinal tract in different animal species (A. mellifera, Litopenaeus vannamei, Mus musculus, Homo sapiens) have been reported as necessary for survival, since their cooperative behavior increases the vigor of a community [28, 39, 47, 49, 50]. Recent studies in fecal samples of farm animals have revealed the presence of intervening sequences (IVS), which are host-specific and provide a basis for the differentiation of the microorganisms derived from different hosts [51].
\nThe role of microbial communities within a host is important. Given the existing delicate balance of these associations, any type of alterations in the microbiome composition could cause disease in the host organism [6, 12, 39, 45]. Previous studies have revealed that in diseased individuals of different species, the microbial diversity is significantly reduced. This could be due to the fact that alterations in the microbiome composition skew the association between the host and the microbiome producing dysbiosis and increasing the number of opportunistic pathogens [6, 12, 13, 16, 27, 39, 45, 52]. In marine environments, the continual presence of pathogens has been observed in environmental samples [16, 44] and in several marine organisms (L. vannamei and M. nipponense) [17, 18, 39, 45, 46]. The continual presence of pathogens in low proportion has been reported during the life cycle of these species, suggesting an active in situ infection in which the host has co-evolved with the parasitic organisms and developed mechanisms that cope with the pathogenic mechanisms of the parasites [13, 16, 17, 27, 45, 46, 52]. It has been observed that the developmental stage in L. vannamei influences the pathogenic response to Vibrio, where the proportion of protective commensal bacteria, Bacteroides and Propionibacterium, tend to decrease as the host aged in contrast the presence of Vibrio increases in diseased individuals [18]. Other mechanisms of coevolution have shown that processes of parasitism and predation can influence the global exchange of resources in an ecosystem. Studies conducted in Escherichia coli and the bacterial predator Myxococcus xanthus have shown that the genome evolution of the predator and prey exhibited accelerated genome evolution when compared to controls, where the predator (M. xanthus) showed adaptations to cell mucoidy and the prey (E. coli) showed adaptations to outer membrane-proteases [7].
\nThe functional analysis of the microbial communities has been carried out using the PICRUST program. This program estimates the families of genes present in a metagenome, by the phylogenetic comparison with sequences of gene families previously reported in databases. These predictions are pre-calculated for genes that code for proteins present in orthologous gene families (COG) or in the Kyoto Encyclopedia of Genes and Genomes (KEEG) [53]. The differential expression of these predicted functions could be assessed with the STAMP software which allows several statistical analysis, size effect, and sample corrections [54]. The use of the afore mentioned protocols have allowed the observation of various attributes in environmental samples related with carbon fixation, amino acid metabolism, and signal transduction in lakes, swamps, and other water bodies [9, 10, 16, 22, 44, 55]. These reports also showed the presence of several bacterial taxa (Actinobacteria, Verrucomicrobia, and Proteobacteria) who were able to synthesize several extracellular enzymes that digests the organic matter [9, 16, 24] or mineralize other nutrients [22, 44].
\nThe influence of the microbiome on the host function have been proposed as a co-evolutionary process where the functionality and the composition of the microbiome can be influenced by the feeding habits of the host [4, 21], and the host can take advantage of the specialized microorganisms who are able to synthesize metabolites that are not present originally in the environment [6, 39]. The consumption of seaweeds by Japanese allows the introduction of algae associated bacteria, which transfer the genes involved in the degradation of the algal sulphated polysaccharides to competent gut resident bacteria with a process known as horizontal gene transfer [28]. Certain marine invertebrates (Elysia chlorotica) that feed on algae are able to maintain the algal plastids as photosynthetically symbionts which allow the use of photosynthates as food source [26]. These examples of coevolutionary processes show how the functionality of the microbiome could be influenced by the dietary habits of the host since; these metabolic add-ons allow the host to thrive in otherwise adverse environmental conditions (oligotrophic habitats).
\nThe metatranscriptomic allows the establishment of parallel relations between the host and the microbiome, but studies require a series of previous steps in order to obtain unbiased information such as the removal of rRNA and the microbial mRNA enrichment (Figure 2) [17, 19, 20, 21, 56].
\nTranscriptomic and metatranscriptomic techniques for the analysis in different samples.
The assembly of genomes and transcriptomes uses short sequences that are separated into fragments known as k-mers, which are aligned and compared graphically (De Brujin graphs) in order to perform de novo reconstruction of the genome or transcriptome. Several programs such as Velvet, SOAP, Trinity, and FLASH are capable of performing it by using a reference genome or transcriptome, if available [8, 57, 58, 59, 60, 61, 62, 63]. In the case of the Trinity platform, it is capable not only of assembling but also of mapping within the assembly (Bowtie1 and Bowtie2), basic statistical analysis of the assembly, quantifying transcripts (RSEM, Salmon, eXpress, and Kallisto), and performing differential expression of transcripts (edgeR, DESeq2, ROTS, and lima/voom) [8].
\nThe metatranscriptomic studies have allowed to reveal the functions of the microorganisms within a host or in different environments and to identify, in both host and microbiome, transcripts related mainly to metabolic processes associated with the nutrient uptake. These observations suggest that the symbiotic chemoautotrophic bacteria provide organic compounds to the host organism that uses it for its nutrition [11, 20, 45, 64]. In fact, recent studies have reported that more than a third of genes are shared among living organisms, especially to those related to the central metabolic pathways (Glucolysis, TCA, Oxidative phosphorylation, Purine and Pyrimidine metabolism) which could increase the efficiency for the digestion of several biomolecules [11, 17, 26, 45].
\nThe use of bioinformatic tools in metatranscriptomics studies has allowed the visualization of the host-microbiome interactions, especially those related with the primary metabolism [13, 38, 52]. The visualization of the shared enzymatic modules is accomplished through the use of identifiers derived from KEGG orthology (KO) and Enzyme Codes (EC) on the iPath3 platform [65]. In this platform, it is possible to overlap metabolic functions (host-symbiont) using the EC and KO identifiers in different metabolic maps (general metabolic pathways, bacterial metabolism, and secondary metabolism), showing graphically the enzymatic modules of each individual and highlighting the enzymatic modules with a shared function.
\nMetatranscriptomic studies have been able to show that microorganisms are capable of generating complex trophic networks communicating with each other through chemical signals in a process known as quorum sensing [12, 26, 56]; however, this process is not restricted only to microorganisms; recent studies have suggested an interdominion quorum sensing [4, 17, 21].
\nThe bioinformatic tools mentioned in this chapter are open-source programs, requiring the user to have a UNIX or OSx operating system installed; a RAM memory greater than 16 GB, a hard disk greater than 500 GB of storage and knowledge about command lines in UNIX [16, 19, 20, 22, 24, 30, 31, 32, 35, 46, 48, 51]. These requirements can be complicated for those who want to initiate a bionformatic analysis; however, there are other options, such as the metaservers, which can allow the data processing in a graphical environment.
\nThe metaservers are web service providers that assemble a series of programs and applications that otherwise are dispersed. Among the most used metaservers are Galaxy, TRUFA, and MG-RAST [22, 24, 34].
\nGalaxy: it is a collaborative initiative that provides a free set of tools and bioinformatics programs ranging from quality control of sequences (FASTQC), sequence editors, data grouping tools, tools for assembly (Trinity), sequence mapping (Bowtie), transcript quantification (Salmon and Kallisto), and metagenomic analysis programs (Mothur, Vegan, Kraken, and Krona) [34]. Being an open initiative, Galaxy presents a series of servers that offer different programs such as the functional prediction of a metagenome by PICRUST (Langille Lab and Huttentowe Lab) and servers dedicated to the functional annotation of transcriptomes (ANASTASIA).
\nTRUFA: Transcriptome User-Friendly Analysis [22], a program developed by the Institute of Physics of Cantabria, is a free server and contains several programs exclusively for transcriptomic (metatranscriptomic) analysis ranging from quality control (FASTQC and PRINSEQ), edited of sequences (CutAdapt), assembly of sequences (Trinity), quantification of transcripts (RSEM and eXpress) and functional annotation (BLAST2GO and HMMER). The files can be edited beforehand, and certain modules of the platform can be accessed, such as the functional annotation in case of already assembled sequences.
\nMG-RAST: Metagenomic Rapid Annotation-based on Subsystems Technology [24] is an open platform capable of analyzing sequences from different NGS platforms (Illumina, PacBio, and Nanopore). Unlike the aforementioned servers, MG-RAST has a pipeline that includes the quality control of the sequences, removal of adapters, detection of isoforms of transcripts, taxonomic comparison, and functional assignment. This server has several databases where the results can be analyzed regarding function (SEED, KEEG, COG, and NOG) and taxonomy (ITS, SILVA, RDP, and GreenGenes). It also has tools to export the data in tabular format, fasta, or in the form of BIOM type matrix.
\nBLAST2GO: it is a sequence annotator that is able to perform searches in the NCBI, which in its basic version has the BLAST algorithms to add taxonomic filters in order to accelerate the annotation. It also allows searches of Interprotein domains (InterProScan), allows the classification of proteins based on the Gene Orthology (GO) database, interaction maps between each GO term, function enrichment analysis (Fisher Exact Test) and the analysis of the metabolic modules present in the KEGG. The PRO version of this program allows making several annotations at the same time, using CLOUD-BLAST services and performing other types of analysis such as the differential expression of transcripts [37].
\nThe study of how microbial communities contribute to environmental functions and the physiology of the host was limited to cultivable microorganisms. As, the free culture techniques based on molecular markers developed (AFLP, RFLP, and DGGE), this knowledge expanded. However, the knowledge obtained from these was quite limited. But now with the techniques of massive sequencing, it has been possible to obtain a better understanding of the role of microorganisms in different types of environments and hosts, both from the taxonomic and functional point of view.
\nThe use of bioinformatic programs has allowed not only the reconstruction of the molecular phylogeny but also has allowed them to be studied from the functional point of view, showing the great potential for the future biotechnological exploitation of this microbial metabolic diversity such as enzymes with different catalytic activities from uncultivable organisms. Several combined methodologies that uses NGS techniques along with culture-based methods have been used to obtain new bacterial strains using specific culture media or through the functional screening using specific primers in order to isolate the genes of interest.
\nCurrent research is now been carried out in order to obtain more precise metagenomic and metatranscriptomic assemblages, with new software specially designed (MetaVelvet, TriMetAss, and MetaAmos) to obtain complete genomes and transcriptomes of the bacterial communities. These protocols along with the integration of other “omic” techniques and systems biology could allow a better understanding of the complex metabolic and trophic networks that operates in an organism or environment.
\nThe authors would like to acknowledge the General Direction of Academic Personal Affairs (DGAPA, UNAM) for the postdoctoral grant for REVG, the National Council of Science and Technology (CONACyT) for funding this project, AquaPacific (Mazatlan, Sinaloa, Mexico) especially to: Bruno Gomez-Gil, Sonia Soto and Daniel Palacios for providing the biological material collection.
\nThe author declares that this chapter was conducted in the absence of any financial or commercial relationships that could be construed as a potential conflict of interest.
The Internet has irrevocably changed the dynamics of scholarly communication and publishing. Consequently, we find it necessary to indicate, unambiguously, our definition of what we consider to be a published scientific work.
",metaTitle:"Prior Publication Policy",metaDescription:"Prior Publication Policy",metaKeywords:null,canonicalURL:"/page/prior-publication-policy",contentRaw:'[{"type":"htmlEditorComponent","content":"A significant number of working papers, early drafts, and similar work in progress are openly shared online between members of the scientific community. It has become common to announce one’s own research on a personal website or a blog to gather comments and suggestions from other researchers. Such works and online postings are, indeed, published in the sense that they are made publicly available. However, this does not mean that if submitted for publication by IntechOpen they are not original works. We differentiate between reviewed and non-reviewed works when determining whether a work is original and has been published in a scholarly sense or not.
\\n\\nThe significance of Peer Review cannot be overstated when it comes to defining, in our terms, what constitutes a published scientific work. Peer Review is widely considered to be the cornerstone of modern publishing processes and the key value-adding contribution to a scholarly manuscript that a publisher can make.
\\n\\nOther than the issue of originality, research misconduct is another major issue that all publishers have to address. IntechOpen’s Retraction & Correction Policy and various publication ethics guidelines identify both redundant publication and (self)plagiarism to fall within the definition of research misconduct, thus constituting grounds for rejection or the issue of a Retraction if the work has already been published.
\\n\\nIn order to facilitate the tracking of a manuscript’s publishing history and its development from its earliest draft to the manuscript submitted, we encourage Authors to disclose any instances of a manuscript’s prior publication, whether it be through a conference presentation, a newspaper article, a working paper publicly available in a repository or a blog post.
\\n\\nA note to the Academic Editor containing detailed information about a submitted manuscript’s previous public availability is the preferred means of reporting prior publication. This helps us determine if there are any earlier versions of a manuscript that should be disclosed to our readers or if any of those earlier versions should be cited and listed in a manuscript’s references.
\\n\\nSome basic information about the editorial treatment of different varieties of prior publication is laid out below:
\\n\\n1. CONFERENCE PAPERS & PRESENTATIONS
\\n\\nGiven that conference papers and presentations generally pass through some sort of peer or editorial review, we consider them to be published in the accepted scholarly sense, particularly if they are published as a part of conference proceedings.
\\n\\nAll submitted manuscripts originating from a previously published conference paper must contain at least 50% of new original content to be accepted for review and considered for publication.
\\n\\nAuthors are required to report any links their manuscript might have with their earlier conference papers and presentations in a note to the Academic Editor, as well as in the manuscript itself. Additionally, Authors should obtain any necessary permissions from the publisher of their conference paper if copyright transfer occurred during the publishing process. Failure to do so may prevent Us from publishing an otherwise worthy work.
\\n\\n2. NEWSPAPER & MAGAZINE ARTICLES
\\n\\nNewspaper and magazine articles usually do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense. Articles appearing in newspapers and magazines rarely possess the depth and structure characteristic of scholarly articles.
\\n\\nSubmitted manuscripts stemming from a previous newspaper or magazine article will be accepted for review and considered for publication. However, Authors are strongly advised to report any such publication in an accompanying note to the External Editor.
\\n\\nAs with the conference papers and presentations, Authors should obtain any necessary permissions from the newspaper or magazine that published the work, and indicate that they have done so in a note to the External Editor.
\\n\\n3. GREY LITERATURE
\\n\\nWhite papers, working papers, technical reports and all other forms of papers which fall within the scope of the ‘Luxembourg definition’ of grey literature do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense.
\\n\\nAlthough such papers are regularly made publicly available via personal websites and institutional repositories, their general purpose is to gather comments and feedback from Authors’ colleagues in order to further improve a manuscript intended for future publication.
\\n\\nWhen submitting their work, Authors are required to disclose the existence of any publicly available earlier drafts in a note to the Academic Editor. In cases where earlier drafts of the submitted version of the manuscript are publicly available, any overlap between the versions will generally not be considered an instance of self-plagiarism.
\\n\\n4. SOCIAL MEDIA, BLOG & MESSAGE BOARD POSTINGS
\\n\\nWe feel that social media, blogs and message boards are generally used with the same intention as grey literature, to formulate ideas for a manuscript and gather early feedback from like-minded researchers in order to improve a particular piece of work before submitting it for publication. Therefore, we do not consider such internet postings to be publication in the scholarly sense.
\\n\\nNevertheless, Authors are encouraged to disclose the existence of any internet postings in which they outline and describe their research or posted passages of their manuscripts in a note to the Academic Editor. Please note that we will not strictly enforce this request in the same way that we would instructions we consider to be part of our conditions of acceptance for publication. We understand that it may be difficult to keep track of all one’s internet postings in which the researcher´s current work might be mentioned.
\\n\\nIn cases where there is any overlap between the Author´s submitted manuscript and related internet postings, we will generally not consider it to be an instance of self-plagiarism. This also holds true for any co-Author as well.
\\n\\nFor more information on this policy please contact permissions@intechopen.com.
\\n\\nPolicy last updated: 2017-03-20
\\n"}]'},components:[{type:"htmlEditorComponent",content:'A significant number of working papers, early drafts, and similar work in progress are openly shared online between members of the scientific community. It has become common to announce one’s own research on a personal website or a blog to gather comments and suggestions from other researchers. Such works and online postings are, indeed, published in the sense that they are made publicly available. However, this does not mean that if submitted for publication by IntechOpen they are not original works. We differentiate between reviewed and non-reviewed works when determining whether a work is original and has been published in a scholarly sense or not.
\n\nThe significance of Peer Review cannot be overstated when it comes to defining, in our terms, what constitutes a published scientific work. Peer Review is widely considered to be the cornerstone of modern publishing processes and the key value-adding contribution to a scholarly manuscript that a publisher can make.
\n\nOther than the issue of originality, research misconduct is another major issue that all publishers have to address. IntechOpen’s Retraction & Correction Policy and various publication ethics guidelines identify both redundant publication and (self)plagiarism to fall within the definition of research misconduct, thus constituting grounds for rejection or the issue of a Retraction if the work has already been published.
\n\nIn order to facilitate the tracking of a manuscript’s publishing history and its development from its earliest draft to the manuscript submitted, we encourage Authors to disclose any instances of a manuscript’s prior publication, whether it be through a conference presentation, a newspaper article, a working paper publicly available in a repository or a blog post.
\n\nA note to the Academic Editor containing detailed information about a submitted manuscript’s previous public availability is the preferred means of reporting prior publication. This helps us determine if there are any earlier versions of a manuscript that should be disclosed to our readers or if any of those earlier versions should be cited and listed in a manuscript’s references.
\n\nSome basic information about the editorial treatment of different varieties of prior publication is laid out below:
\n\n1. CONFERENCE PAPERS & PRESENTATIONS
\n\nGiven that conference papers and presentations generally pass through some sort of peer or editorial review, we consider them to be published in the accepted scholarly sense, particularly if they are published as a part of conference proceedings.
\n\nAll submitted manuscripts originating from a previously published conference paper must contain at least 50% of new original content to be accepted for review and considered for publication.
\n\nAuthors are required to report any links their manuscript might have with their earlier conference papers and presentations in a note to the Academic Editor, as well as in the manuscript itself. Additionally, Authors should obtain any necessary permissions from the publisher of their conference paper if copyright transfer occurred during the publishing process. Failure to do so may prevent Us from publishing an otherwise worthy work.
\n\n2. NEWSPAPER & MAGAZINE ARTICLES
\n\nNewspaper and magazine articles usually do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense. Articles appearing in newspapers and magazines rarely possess the depth and structure characteristic of scholarly articles.
\n\nSubmitted manuscripts stemming from a previous newspaper or magazine article will be accepted for review and considered for publication. However, Authors are strongly advised to report any such publication in an accompanying note to the External Editor.
\n\nAs with the conference papers and presentations, Authors should obtain any necessary permissions from the newspaper or magazine that published the work, and indicate that they have done so in a note to the External Editor.
\n\n3. GREY LITERATURE
\n\nWhite papers, working papers, technical reports and all other forms of papers which fall within the scope of the ‘Luxembourg definition’ of grey literature do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense.
\n\nAlthough such papers are regularly made publicly available via personal websites and institutional repositories, their general purpose is to gather comments and feedback from Authors’ colleagues in order to further improve a manuscript intended for future publication.
\n\nWhen submitting their work, Authors are required to disclose the existence of any publicly available earlier drafts in a note to the Academic Editor. In cases where earlier drafts of the submitted version of the manuscript are publicly available, any overlap between the versions will generally not be considered an instance of self-plagiarism.
\n\n4. SOCIAL MEDIA, BLOG & MESSAGE BOARD POSTINGS
\n\nWe feel that social media, blogs and message boards are generally used with the same intention as grey literature, to formulate ideas for a manuscript and gather early feedback from like-minded researchers in order to improve a particular piece of work before submitting it for publication. Therefore, we do not consider such internet postings to be publication in the scholarly sense.
\n\nNevertheless, Authors are encouraged to disclose the existence of any internet postings in which they outline and describe their research or posted passages of their manuscripts in a note to the Academic Editor. Please note that we will not strictly enforce this request in the same way that we would instructions we consider to be part of our conditions of acceptance for publication. We understand that it may be difficult to keep track of all one’s internet postings in which the researcher´s current work might be mentioned.
\n\nIn cases where there is any overlap between the Author´s submitted manuscript and related internet postings, we will generally not consider it to be an instance of self-plagiarism. This also holds true for any co-Author as well.
\n\nFor more information on this policy please contact permissions@intechopen.com.
\n\nPolicy last updated: 2017-03-20
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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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After obtaining a Master's degree in Mechanical Engineering, he continued his PhD studies in Robotics at the Vienna University of Technology. Here he worked as a robotic researcher with the university's Intelligent Manufacturing Systems Group as well as a guest researcher at various European universities, including the Swiss Federal Institute of Technology Lausanne (EPFL). During this time he published more than 20 scientific papers, gave presentations, served as a reviewer for major robotic journals and conferences and most importantly he co-founded and built the International Journal of Advanced Robotic Systems- world's first Open Access journal in the field of robotics. Starting this journal was a pivotal point in his career, since it was a pathway to founding IntechOpen - Open Access publisher focused on addressing academic researchers needs. Alex is a personification of IntechOpen key values being trusted, open and entrepreneurial. Today his focus is on defining the growth and development strategy for the company.",institutionString:null,institution:{name:"TU Wien",country:{name:"Austria"}}},{id:"19816",title:"Prof.",name:"Alexander",middleName:null,surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/19816/images/1607_n.jpg",biography:"Alexander I. Kokorin: born: 1947, Moscow; DSc., PhD; Principal Research Fellow (Research Professor) of Department of Kinetics and Catalysis, N. Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow.\r\nArea of research interests: physical chemistry of complex-organized molecular and nanosized systems, including polymer-metal complexes; the surface of doped oxide semiconductors. He is an expert in structural, absorptive, catalytic and photocatalytic properties, in structural organization and dynamic features of ionic liquids, in magnetic interactions between paramagnetic centers. The author or co-author of 3 books, over 200 articles and reviews in scientific journals and books. He is an actual member of the International EPR/ESR Society, European Society on Quantum Solar Energy Conversion, Moscow House of Scientists, of the Board of Moscow Physical Society.",institutionString:null,institution:{name:"Semenov Institute of Chemical Physics",country:{name:"Russia"}}},{id:"62389",title:"PhD.",name:"Ali Demir",middleName:null,surname:"Sezer",slug:"ali-demir-sezer",fullName:"Ali Demir Sezer",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/62389/images/3413_n.jpg",biography:"Dr. Ali Demir Sezer has a Ph.D. from Pharmaceutical Biotechnology at the Faculty of Pharmacy, University of Marmara (Turkey). He is the member of many Pharmaceutical Associations and acts as a reviewer of scientific journals and European projects under different research areas such as: drug delivery systems, nanotechnology and pharmaceutical biotechnology. Dr. Sezer is the author of many scientific publications in peer-reviewed journals and poster communications. Focus of his research activity is drug delivery, physico-chemical characterization and biological evaluation of biopolymers micro and nanoparticles as modified drug delivery system, and colloidal drug carriers (liposomes, nanoparticles etc.).",institutionString:null,institution:{name:"Marmara University",country:{name:"Turkey"}}},{id:"61051",title:"Prof.",name:"Andrea",middleName:null,surname:"Natale",slug:"andrea-natale",fullName:"Andrea Natale",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"100762",title:"Prof.",name:"Andrea",middleName:null,surname:"Natale",slug:"andrea-natale",fullName:"Andrea Natale",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"St David's Medical Center",country:{name:"United States of America"}}},{id:"107416",title:"Dr.",name:"Andrea",middleName:null,surname:"Natale",slug:"andrea-natale",fullName:"Andrea Natale",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Texas Cardiac Arrhythmia",country:{name:"United States of America"}}},{id:"64434",title:"Dr.",name:"Angkoon",middleName:null,surname:"Phinyomark",slug:"angkoon-phinyomark",fullName:"Angkoon Phinyomark",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/64434/images/2619_n.jpg",biography:"My name is Angkoon Phinyomark. I received a B.Eng. degree in Computer Engineering with First Class Honors in 2008 from Prince of Songkla University, Songkhla, Thailand, where I received a Ph.D. degree in Electrical Engineering. My research interests are primarily in the area of biomedical signal processing and classification notably EMG (electromyography signal), EOG (electrooculography signal), and EEG (electroencephalography signal), image analysis notably breast cancer analysis and optical coherence tomography, and rehabilitation engineering. I became a student member of IEEE in 2008. During October 2011-March 2012, I had worked at School of Computer Science and Electronic Engineering, University of Essex, Colchester, Essex, United Kingdom. In addition, during a B.Eng. I had been a visiting research student at Faculty of Computer Science, University of Murcia, Murcia, Spain for three months.\n\nI have published over 40 papers during 5 years in refereed journals, books, and conference proceedings in the areas of electro-physiological signals processing and classification, notably EMG and EOG signals, fractal analysis, wavelet analysis, texture analysis, feature extraction and machine learning algorithms, and assistive and rehabilitative devices. I have several computer programming language certificates, i.e. Sun Certified Programmer for the Java 2 Platform 1.4 (SCJP), Microsoft Certified Professional Developer, Web Developer (MCPD), Microsoft Certified Technology Specialist, .NET Framework 2.0 Web (MCTS). 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