Number of ovules with different types of embryo sacs at anthesis in apomictic varieties of
\\n\\n
Released this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\\n\\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
Note: Edited in March 2021
\\n"}]',published:!0,mainMedia:{caption:"Highly Cited",originalUrl:"/media/original/117"}},components:[{type:"htmlEditorComponent",content:'IntechOpen is proud to announce that 191 of our authors have made the Clarivate™ Highly Cited Researchers List for 2020, ranking them among the top 1% most-cited.
\n\nThroughout the years, the list has named a total of 261 IntechOpen authors as Highly Cited. Of those researchers, 69 have been featured on the list multiple times.
\n\n\n\nReleased this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\n\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
Note: Edited in March 2021
\n'}],latestNews:[{slug:"webinar-introduction-to-open-science-wednesday-18-may-1-pm-cest-20220518",title:"Webinar: Introduction to Open Science | Wednesday 18 May, 1 PM CEST"},{slug:"step-in-the-right-direction-intechopen-launches-a-portfolio-of-open-science-journals-20220414",title:"Step in the Right Direction: IntechOpen Launches a Portfolio of Open Science Journals"},{slug:"let-s-meet-at-london-book-fair-5-7-april-2022-olympia-london-20220321",title:"Let’s meet at London Book Fair, 5-7 April 2022, Olympia London"},{slug:"50-books-published-as-part-of-intechopen-and-knowledge-unlatched-ku-collaboration-20220316",title:"50 Books published as part of IntechOpen and Knowledge Unlatched (KU) Collaboration"},{slug:"intechopen-joins-the-united-nations-sustainable-development-goals-publishers-compact-20221702",title:"IntechOpen joins the United Nations Sustainable Development Goals Publishers Compact"},{slug:"intechopen-signs-exclusive-representation-agreement-with-lsr-libros-servicios-y-representaciones-s-a-de-c-v-20211123",title:"IntechOpen Signs Exclusive Representation Agreement with LSR Libros Servicios y Representaciones S.A. de C.V"},{slug:"intechopen-expands-partnership-with-research4life-20211110",title:"IntechOpen Expands Partnership with Research4Life"},{slug:"introducing-intechopen-book-series-a-new-publishing-format-for-oa-books-20210915",title:"Introducing IntechOpen Book Series - A New Publishing Format for OA Books"}]},book:{item:{type:"book",id:"10162",leadTitle:null,fullTitle:"A Diffusion Hydrodynamic Model",title:"A Diffusion Hydrodynamic Model",subtitle:null,reviewType:"peer-reviewed",abstract:"The Diffusion Hydrodynamic Model (DHM), as presented in the 1987 USGS publication, was one of the first computational fluid dynamics computational programs based on the groundwater program MODFLOW, which evolved into the control volume modeling approach. Over the following decades, others developed similar computational programs that either used the methodology and approaches presented in the DHM directly or were its extensions that included additional components and capacities. Our goal is to demonstrate that the DHM, which was developed in an age preceding computer graphics/visualization tools, is as robust as any of the popular models that are currently used. We thank the USGS for their approval and permission to use the content from the earlier USGS report.",isbn:"978-1-83962-818-4",printIsbn:"978-1-83962-817-7",pdfIsbn:"978-1-83962-819-1",doi:"10.5772/intechopen.90224",price:100,priceEur:109,priceUsd:129,slug:"a-diffusion-hydrodynamic-model",numberOfPages:88,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"a8c90b653db4fa7a59132d39cca185d8",bookSignature:"Theodore V. 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The model results, for different inflow scenarios, are compared with the standard United States Geological Survey (USGS) K-634 model. The sensitivity of the model results to grid spacing and the chosen time step are presented. The model results are in close agreement.",signatures:"Theodore V. Hromadka II and Chung-Cheng Yen",downloadPdfUrl:"/chapter/pdf-download/72798",previewPdfUrl:"/chapter/pdf-preview/72798",authors:[{id:"181008",title:"Dr.",name:"Theodore V.",surname:"Hromadka II",slug:"theodore-v.-hromadka-ii",fullName:"Theodore V. Hromadka II"}],corrections:null},{id:"72808",title:"Program Description of the Diffusion Hydrodynamic Model",doi:"10.5772/intechopen.93209",slug:"program-description-of-the-diffusion-hydrodynamic-model",totalDownloads:340,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"The numerical algorithm, with a focus on the interface element that is used in the diffusion hydrodynamic model, is presented in this chapter. The source program was written in FORTRAN language, and it can be downloaded from this book companion website. The channel, flood plain, and the interface flow details are discussed.",signatures:"Theodore V. Hromadka II and Chung-Cheng Yen",downloadPdfUrl:"/chapter/pdf-download/72808",previewPdfUrl:"/chapter/pdf-preview/72808",authors:[{id:"181008",title:"Dr.",name:"Theodore V.",surname:"Hromadka II",slug:"theodore-v.-hromadka-ii",fullName:"Theodore V. Hromadka II"}],corrections:null},{id:"72824",title:"Applications of Diffusion Hydrodynamic Model",doi:"10.5772/intechopen.93210",slug:"applications-of-diffusion-hydrodynamic-model",totalDownloads:546,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"The diffusion hydrodynamic model is applied for seven engineering applications that are commonly encountered in real-world applications. Of the seven applications, six relate to two-dimensional flows. The results are compared to other model results, where available. 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His scientific work mainly involves cardiac arrhythmias, heart failure, cardiomyopathies, and implantable cardiac devices. Dr. Magnusson is a renowned speaker and leader of educational activities for diverse groups of professionals.",institutionString:"Karolinska Institute",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"11",totalChapterViews:"0",totalEditedBooks:"3",institution:{name:"Karolinska Institute",institutionURL:null,country:{name:"Sweden"}}}],coeditorOne:{id:"305337",title:"Dr.",name:"Jo Ann",middleName:null,surname:"LeQuang",slug:"jo-ann-lequang",fullName:"Jo Ann LeQuang",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002nKU1EQAW/Profile_Picture_1638351508200",biography:"Jo Ann LeQuang is a medical writer who got her start working in the pacemaker industry when she was hired to work for Intermedics to assist in setting up a translation department for software localization for implantable devices. 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Drug delivery systems are designed to enhance the efficiency and safety of therapeutics by regulating the rate, time and place of release in the body [1, 2]. Drug delivery technology has emerged as an essential tool for the improvement of drug bioavailability, reduction in the side effects of medication, all of which generate remarkable clinical outcomes [3]. Drugs may be administered to the body via local application, enterally or parenterally. The parenteral route typically relates to administration that excludes absorption from the gastrointestinal tract (GIT). It consists of administration by injection, inhalation and via transdermal routes. The enteral route is associated with the absorption of the drug via the GIT, this includes oral, sublingual, and rectal administration. Aptly, the mode of drug administration depends on the disease, the desired therapeutic effect and the nature of the product available [4]. Moreover, each delivery route has inherent benefits and constraints. Nevertheless, the majority of manufactured medicines in the pharmaceutical industry are delivered orally, owing to the distinctive advantages offered by this route, including versatility in accommodating various types of drugs, simplicity of administration and accessibility, patient compliance and safety profiles [5, 6, 7]. Additionally, the intestinal epithelium is an ideal platform for drug absorption due to the viscous mucosal layer lined with abundant enterocytes, goblet cells, and Peyer’s patches that trap drug molecules within the mucus as they transit the GIT. [8, 9].
In comparison to other routes of administration, the oral route is exceptionally complex in expression of anatomical features physiology throughout the GIT [10]. Furthermore, these expressions vary along the GIT in both intensity structure. For example, the mucus layer varies in composition and physical properties along the GIT and the pH varies significantly in the main sections of the GIT. The gastrointestinal motility also varies in intensity and form along the GIT and also depends on food status [11]. Even though, these features can impede drug delivery across the GIT, through careful interplay between formulation science and GIT physiology, scientists have been able to exploit this variance for improved drug delivery. In this regard, nanoparticle formulations have immerged as strong contenders able to surmount some of the constraints associated with GIT absorption. Nanoparticles have gained great interest by researchers in recent years as they can be used to improve drug solubility and bioavailability in the harsh GIT environment due to increased surface area to volume ratio, thus provide a rapid onset of therapeutic action [12]. They can also be used to targeting specific sites within the GIT and hence reduce the effects of enzymatic degradation, all of which can improve the safety and effectiveness of drugs [12, 13].
Nanoparticle formulations may be presented in various forms however, polymeric nanoparticles present the versatility of polymers and can be tailored to achieve superior drug stability, enhanced drug payload capacity, longer circulation times and controlled drug release capabilities, when compared with other their colloidal counterparts [14, 15]. In this regard, chitosan-based nanoparticle formulation have been shown to present several of the desirable attributes listed above in addition to being biodegradable, having low toxicity, amenable to tuneable physical properties and bio-adhesive properties [16, 17].
In this chapter we will be discussing the interplay between the GIT physiology/anatomy and drug physicochemical/biopharmaceutical factors in the absorption process that influence oral therapeutics. We, will also review the physicochemical properties of chitosan relevant for effective GIT delivery, including methods of formulation. The most utilised nanoparticle formulation methods used for chitosan-based nanoparticles are also examined. Finally, we will highlight the recent developments on chitosan-based nanoparticles used in the oral delivery of different drugs.
The GIT, also known as the digestive tract or alimentary canal, is approximately 9 meters long and can be functionally divided into two parts, the upper and the lower GIT (Figure 1). The upper GIT; consisting of mouth, pharynx, oesophagus, stomach and small intestine, play a major role in the transport of the swallowed food bolus, enzymatic digestion and absorption of nutrients [18]. The lower GIT is usually referred to the large intestine and is responsible for the adsorption of water, fermentation of undigested sugars and the storage and evacuation of stool [19]. Following oral dosing, the drug traverses several semipermeable cell membranes through its trajectory to absorption and eventually enters the general circulatory system. Drugs cross cell membranes, which comprise of bimolecular lipid matrix, either by passive diffusion or active transport.
The anatomy of the human gastrointestinal tract.
The most prevalent form of absorption of the majority of orally administered drugs is by passive diffusion across cell membranes. This process comprises of a three-step process, whereby the permeant first transverses into the membrane, disperses across it and then is released into the cytosol [20]. Typically, drug molecules move down a concentration gradient, from a region of high concentration (e.g., GI fluids) to one of low concentration (e.g., blood), without the expenditure of energy [21]. Usually, a concentration gradient is manifested as a disparity in concentration of a substance within an area and is linearly related to the diffusion rate. The latter is also governed by the lipid solubility, size and polarity of the drug species.
Most drugs are either weak acids or bases and occur either in the unionized or ionized form as a function of pH [22]. For lipophilic drugs, the unionized form of drug, may penetrate cell membranes easily as the membrane is lipoidal. On the other hand, hydrophilic drugs, present an ionized form of the drug, which has high electrical resistance and thus cannot traverse the cell membranes easily but may diffuse through the para-cellular spaces. However, it is worth noting that the para-cellular junctions contribute to less than 0.01% of the entire GIT surface area and furthermore, the permeability of these junctures diminishes down the GIT [23]. Additionally, the capability of drugs to traverse a membrane also relies on the acid–base dissociation constant (pKa) of the drug in question. The pKa is the pH at which concentrations of ionized and unionized forms are equivalent [24]. So, if the pH is less than the pKa, the unionized form of a weak acid prevails, and
Overall, the process by which molecules traverse cell membranes is by passive diffusion, down the concentration gradient. However large hydrophilic ionic molecules and charged molecules cannot freely traverse the phospholipid bilayer cell membrane passively. Their transport may be confined to protein channels and distinct transport mechanisms present within the membrane [25]. Such drugs gain access through the membrane by facilitated diffusion whereby molecules integrate with embedded protein carriers to shuttle them across the membrane. This process does not expend energy and is also down the concentration gradient though quicker than would be anticipated by diffusion alone [26]. A frequent case of facilitated diffusion is the migration of glucose into cells during the production of adenosine triphosphate (ATP). Glucose is both large and polar thereby unable to pass the lipid bilayer via simple diffusion. Hence, glucose molecules are delivered into the cell via a unique carrier protein (glucose transporter) to promote its internalisation in cells [27].
Active transport is an energy-dependent process that translocates drug molecules against their concentration gradient by a molecular pump [20]. Carrier-mediated active transport demand energy via ATP hydrolysis or by accompanying the co-transport of counter ions down its electrochemical gradient (e.g., Na+, H+, Cl−) [28]. The most common active transport system is the sodium-potassium pump and receptor-mediated endocytosis. Energy can either be directly provided to the ion pump or indirectly by connecting a pump-action to an activated ionic gradient. It is often encountered in the gut mucosa, the liver, renal tubules and the blood–brain barrier [22]. Active transport is typically restricted to drugs that structurally resemble endogenous substances; e.g., vitamins and amino acids, and that are absorbed via specific sites in the small intestine. Targeting drugs to these transporters can enhance their bioavailability and distribution [21].
The sodium-potassium pump system (Na+/K+ ATPase), utilises ATP to move Na+ and K+ in and out of the cell. It is a vital ion pump located in the membranes of various cell types, such as the Na+/amino acid symport in the mucosal cells of the small bowel [22, 29].
Cells control the endocytosis of certain substances via receptor-mediated endocytosis. The use of this form of endocytosis in the GIT is crucial for oral delivery of drugs because it delays the transit of drugs in GIT. Receptor-mediator endocytosis involves the internalisation of macromolecules by binding the latter to receptors considered as membrane-associated protein [30]. There are more than 20 different receptors involved in the internalisation of macromolecules [31]. Following binding to the receptor on the cell surface, the cell will endocytose the portion of the cell membrane enclosing the receptor-ligand complex via a clathrin-dependent endocytic process [28]. Clathrin plays a significant role in the formation of clathrin-coated pits; invaginated regions of the plasma membrane, and pinch off to form clathrin-coated vesicles that transport molecules within cells [31].
In summary, drug adsorption may occur passively or via active transport. In either case, absorption occurs predominantly in the small intestine due to its more permeable membrane and larger surface area provided by the microvilli. Even though, the stomach has a relatively broad epithelial surface, yet the dense mucus layer and transient transit times expended by dosage forms contribute to an impeded absorption. Moreover, the colon with an absorptive surface area of about 5m2 has negligible contribution to drug absorption in GIT, due to slow caecal arrival times of dosage forms, the presence of numerous gut bacteria and solid stool that impede lateral diffusion. All in all, absorption of oral drugs is interlinked and controlled by various intrinsic factors; like drug solubility, dissolution and permeability across the mucosal barriers, and physiological factors; such as gastrointestinal transit time, pH and gut microbiome [13, 32].
Drug dissolution, solubility and permeability are the three fundamental parameters used in the Biopharmaceutics Classification System (BCS) to predict the factors limiting drug absorption from GIT [33]. The BCS is recognised as a useful tool for designing drug delivery systems and is adopted by the US Food and Drug Administration (FDA), the European Medicines Agency (EMA) and the World Health Organization (WHO) [34]. According to the BCS, all drug substances are classified into four categories: class I—high soluble and high permeable, class II—low soluble and high permeable, class III—low soluble and high permeable and class IV—low soluble and low permeable [35].
Drug solubility is crucial outcome in pharmaceutical dosage form. In the BCS system, a drug is deemed highly soluble when the maximal dose strength is soluble in 250 mL of aqueous media across the pH range of 1 to 7.5 [35]. However, more than 40% of the established new chemical entities in the pharmaceutical sector are considered insoluble in water, causing inadequate bioavailability [36]. This makes solubility amongst the most important rate limiting parameters in GIT absorption.
Drug dissolution reflects a dynamic consequence to drug absorption [33], whereby drug is released, dissolved and made accessible for absorption. With the exception of enteric formulations and drugs with low acid solubility, the dissolution process for majority of drugs starts in the stomach where the volume of gastric fluid is sufficient to attain effective drug dissolution [37]. Thus, the gastric fluid containing the disintegrated immediate-release dosage forms brings the solubilized drug into contact with the absorptive surface of the small intestine as absorption in the stomach is generally minimal.
Drug permeability represents the final frontier in the sequence of rate-liming steps to systemic drug availability. It is a measure of the ease of permeation of the drug across the intestinal wall. There is a positive association between the intestinal permeability and drug solubility GI milieu, which in turn depends on the physicochemical characteristics of the drug [38], including the pKa, particle size, lipophilicity, as discussed in the sections below. The ultimate amount of drug absorbed from the GIT also bears dependence on its transit time in the GIT [39].
The GI pH influences the extent of ionization of drug molecules and thereby impacts on its absorption across the epithelium. Variations in pH across the GIT can be exploited for delayed drug release in desired section of the GIT in order to achieve efficient absorption. The fasted stomach is acidic, with pH range of 1–3, which increases upon food or liquid intake. Food is known to buffer the acidic content of the stomach. A rise in pH resumes in response to the continual gastric secretion and then finally, the pH reverts to the original levels due to gastric emptying of content; [40]. The gastric emptying rate significantly affects the rate of drug absorption because it regulates arrival in the duodenum, where the epithelial surface is suited for absorption [41]. Moreover, the disparity in gastric pH conditions affects the drug delivery behaviour of modified release dosage forms such as enteric coated products, where the onset of release along with the overall release kinetics may be changed [42].
The arrival of orally administered dosage forms into the small intestine is met by a pH of about 6 in the duodenum through to pH 7.4 at the terminal ileum [43]. This high pH variability is due to duodenal secretion of alkaline bicarbonate. During postprandial state, the initial intestinal pH drops due to the influx of acidic chyme, which is buffered by bicarbonate secretion as it travels distally [13]. Besides, the mean pH in proximal small intestine during the first hour of transit is usually 6.6, which is further decreased to 5–6 in the distal duodenum [44].
Typically, the pH in the caecum drops to just below pH 6 owing to the fermentation processes of the colonic microbiota and then rises to pH 7 at the rectum [42]. The drop in the amount of short chain fatty acids at the distal colon causes the secretion of colonic mucosal bicarbonate that leads to a neutral pH. Short chain fatty acids are the end products of fermentation of dietary fibres by the anaerobic intestinal microbiota [45]. As a consequence of the neutral pH of the colonic luminal fluid, the solubilisation of drug is the rate-limiting factor in colonic drug absorption [46]. The unspecific interactions of drugs with colonic content (e.g. dietary residues, intestinal secretions or faecal matter) all adds to the odds of effective adsorption across the colon [47].
Generally, the GIT transit time of most orally administered doses through buccal cavity and oesophagus is transient. The stomach is naturally the first segment of the GIT, wherein disintegration and dissolution of solids such as drugs and formulations occur [42]. The period required for a dosage form to exit the stomach is inconstant and relies on several physiological factors, such as age, body posture, gender and food intake [48]. Gastric transit can span from 0 to 2 h in the fasted state and can be extended up to 6 h after food intake [47]. The small intestine is the region of choice for drug absorption with a transit time ranging from 2 to 6 h in healthy individuals. The dissolution of poorly soluble, weakly acidic compounds and lipophilic compounds is greatly enhanced in this region [13]. In colon-specific drug delivery, the drug has to cross the whole GIT prior to arrival at the colon. Thus, the transit time across the colon can be highly variable, and ranges from 20 to 56 h in healthy humans, although higher variations are also reported in literature amounting up to 72 h [42, 49, 50]. Variations in colonic transit time are affected by dosing time, bowel movements as well as gender, whereby females generally have longer colonic transit times than males [51, 52].
Enzymatic and microbial degradation of GIT content affects the amount ultimately made available for absorption. The active sites for most endogenous enzymes are the stomach and small intestine. Even though these enzymes may affect the stability of orally administered drugs, it is possible to exploit this property for regional drug delivery of formulations in the GIT [47]. On the other hand, the intestinal microbiome which includes 500–1000 bacterial species is also important for the digestion of food and the metabolism of drugs [53]. Gastrointestinal microbiome is found in both upper and lower GIT, whereby, a lower bacterial number (1013–1014 bacteria mL−1 of intestinal content) is in the upper GIT because of the fast luminal flow, intestinal fluid volume, and the secretion of bactericidal compounds in this part of the GIT, and highest bacterial community (1010–1011 bacteria mL−1 of intestinal content) is in the colon, in which the redox potential is low and the transit time is long [54, 55]. Therefore, greater number of the intestinal microbiome exists in the anaerobic colon, in which the fermentation of carbohydrates contributes to their nourishment. Usually, orally administered drugs are transformed to bioactive, bio-inactive, or toxic metabolites by the gut microbial population, all of which can impede the bioavailability of drug. However, gut microflora can improve drug bioavailability by eliminating polar moiety from derived conjugates and thereby promoting biliary recycling of compounds [13].
Thus, formulation scientist must be cognizant of the interplay between drug and physiological and anatomical manifestations within the GIT when designing orally administered dosage forms. For example, enteric coating can be applied to dosage forms to delay the release of the API in the acidic gastric fluid until pH above 5.0 [56]. Enteric coating may also be used to shield acid-labile drugs from gastric distress, and upon arrival to the alkaline pH milieu, the enteric polymer coating disintegrates within the intestinal fluid, releasing the drug [57]. Despite employing such coatings and other conventional interventions, numerous pharmaceuticals still display insufficient bioavailability through the oral route of administration. This necessitates the use of alternate strategies. One area of research that is gaining traction more recently is the employment of nanoparticles.
Nanoparticle technology is a multidisciplinary field that utilizes principles from chemistry, biology, physics and engineering to design and fabricate submicronic (< 1 μm) colloidal systems [58]. Nanotechnology has several pharmaceutical and medical applications wherein nanoparticles (NPs), with sizes comparable to large biological molecules such as enzymes can be employed in the delivery of therapeutic agents [59]. The effectiveness of the nanoscale drug delivery vehicles lies on their ability to attain the following key attributes [60]:
The NP must be able to bind or contain the appropriate drug.
The nanocarrier must stay stable in the serum to allow systemic delivery of the therapeutics and only release the drug once at the required site.
The NP-drug complex has to reach the required site either via receptor-mediated interactions or by the enhanced permeability and retention (EPR) effect.
The residual NP carrier should ideally be made of a biological or biologically inert material with a limited lifespan to allow safe degradation.
There are several types of NP drug delivery systems, which may be broadly divided as organic and inorganic NPs [61]. Their particle size, surface charge (ζ potential), hydrophilicity/hydrophobicity, composition, etc. can be tailored for a diverse applications [62]. The primary consideration when designing orally administered NP drug delivery system is to maximise drug concentration in the GI therapeutic window.
Organic NP (Figure 2) are solid particles comprised of organic compounds (usually lipidic or polymeric) ranging from 10 nm to 1 μm [63]. They can be formulated by simple techniques to encapsulate therapeutic agents. Preferably, compounds used in formulation of organic NPs should be biodegradable and biocompatible [61]. Manifestations of organic NP include liposomal, polymeric and solid lipid NP, each system possessing requisite features that addresses physiological and anatomic constraints addressed in sections above. In addition, others systems such as micelles, dendrimers etc. have been also explored as effective nanocarriers for effective deployment of APIs in the GIT [14, 64].
Examples of organic nanoparticle platforms for drug delivery.
Inorganic NP represent a wide spectrum of systems synthesized from metals, metal oxides, and metal sulphides [65]. Gold, silica and superparamagnetic oxide NP are among the long list of inorganic NP (Figure 3). They have been studied for use in imaging on nuclear magnetic resonance and high-resolution superconducting quantum interference devices, and their intrinsic properties have been utilised for therapy [66]. Inorganic NP can easily be conjugated to ligands for tumour targeting and/or with chemotherapeutics for tumour therapy. Additionally, their surface composition can be feasibly manipulated to create NP that can escape the reticuloendothelial system [67]. Even though inorganic NP present good stability characteristics, they have not been the focus of attention in oral NP research, possibly due to concerns on the degradation and elimination end products, which can be potentially toxic [68].
Examples of inorganic NP platforms for drug delivery.
Generally, inorganic NPs differ conceptually from organic NPs in terms of fabrication principles. Inorganic NPs can be formed by the precipitation of inorganic salts, which are linked within a matrix, whilst, most organic NPs are formed by several organic molecules through self-organization or chemical binding [61]. Notwithstanding, both types of NP are very promising in the formulation of oral delivery system and forms part of the evolutional success in several clinical applications. Polymeric NP arguably presents more desirable attributes as orally delivered NP because of their higher stability, enhanced drug payload and controlled drug release capabilities compared with their colloidal counterparts [14, 69].
According to Alexis F. et al., polymeric NP represent the most effective nanocarrier system for prolonged drug delivery [70]. ‘Polymeric NPs’ include any type of polymer formed as NP. Nanospheres are solid spherical NP with molecules attached or adsorbed to their surface, whilst nanocapsules are vesicular systems with substances confined within a cavity consisting of a liquid core (either water or oil) surrounded by a solid shell [71]. Characteristic properties of polymers such as molecular weight, hydrophobicity and crystallinity can be explored to manifest controlled drug release kinetics and entrapment of therapeutic agents [72]. Polymers also provide significant flexibility in the design of oral NP and many exhibit biodegradability [73]. In this regard, synthetic and natural variants have been studied. For example poly-lactic-co-glycolic-acid (PLGA) and poly-lactic-acid (PLA) are synthetic whilst natural polymers include gelatine, dextran, and chitosan [74]. The use of natural polymers is preferred over the synthetic ones as the former usually exhibit less toxicity, widely available and have lower production costs [75]. Chitosan is arguable one of the most studied polymer in NP formulation in view of its distinctive properties. In orally administered NP, chitosan offers added desirability including muco-adhesiveness, augmenting the dissolution rate of poorly water-soluble drugs; useful in drug targeting in the GIT [76].
Chitosan is a hydrophilic, cationic polysaccharide soluble in dilute acids such as acetic acid and formic acid, due to protonated amine groups (NH3+) [75]. It is an N-acetylated derivative of chitin, a natural polysaccharide found in the shells of marine crustaceans. Chitin is chemically inert and thus has fewer applications that chitosan [77]. The acetamido group of chitin, (C2H4NO) can be turned into amino group to yield chitosan by the alkaline deacetylation of chitin. Chitosan is approved as safe by the United States Food and Drug Administration (US-FDA) for dietary use and wound dressing applications, but its toxicity increases with electrical charge and degree of deacetylation [17]. Chemically, it comprises of β- [1–4] -linked D-glucosamine and N-acetylated units (Figure 4).
Chemical structure of chitosan, comprising N-acetyl-D-glucosamine (right) and D-glucosamine (left) units.
The amine group has pKa of 6.2–6.5 [78]. At slightly acidic pH values, the amine groups (NH3+) become protonated, hence possessing the ability to effectively form electrostatic interactions with negatively charged species within mucin in the GIT [75]. Positively charged moieties of chitosan also interact with the tight junctions of the intestinal epithelial cells and thus modulate drug permeation and absorption through the interstitial space between epithelial cells [79]. Moreover, the existence of both hydroxyl and amino groups offers various possibilities for chemical modification. Chemical modifications give rise to different functional derivatives of chitosan like carboxylation, thiolation, alkylation, acylation etc. that further imparts desirable physiochemical and biopharmaceutical properties, such as solubility, adsorption and pH sensitivity in oral drug delivery [80]. For example, N-trimethyl chitosan chloride is developed to amplify the intestinal solubility of chitosan; thiolated chitosan is produced to augment the mucoadhesiveness of chitosan; quaternization of chitosan reinforces its impact on the tight junctions of the GIT epithelium whilst grafting carboxylated chitosan with poly(methyl methacrylate) imparts increased pH sensitivity [81]. Physical modification through blending with other polymers may be used to enhance desirable physical properties. For example, blending of chitosan with polyethylene glycol (PEG) and polyvinyl alcohol (PVA) ameliorate the hydrophilic property of chitosan, while blending of chitosan with cellulose improves its antibacterial properties [82].
Some of the key desirable features in orally administered dosage forms is delayed GI transit in the duodenum and ability to traverse the epithelium effectively. In this regard, chitosan-based NP have been shown to possess these attributes. Mucoadhesion refers to the adhesion between two materials, one of which is mucosal [83]. It can be utilised to prolong the GI transit of dosage forms in the duodenum, thereby improving bioavailability. Delayed transit results from interactions of positively charged moieties in chitosan with negatively charged moieties in sialic acid within mucin [81]. Chitosan is also capable of physically penetrating the mucous network. Prolonged GI residence results in higher net drug flux across the GIT membrane. Drug flux is a combination of passive diffusion and uptake of whole NP by Peyer’s patches [84]. Moreover, chitosan offers controlled drug release capabilities via diffusion from the matrix. Yin et al. prepared thiolated trimethyl chitosan NP for the oral delivery of insulin, where increase in the mucoadhesion resulted in increased insulin transport through rat intestine and uptake by Peyer’s patches compared to controls. They attributed these results to the disulfide bond formation between the NP and mucin [85]. Overall, to achieve the desired properties of interest such as particle size, particle size distribution and area of application, the mode of preparation of chitosan NP plays an essential role.
The preparation of chitosan NP is principally divided into two approaches. The first approach is based on a two-step procedure, where an emulsification system is carried out to generate nanodroplets in which organic compounds (polymer, monomer, and lipid) are solubilized, followed by precipitation or polymerisation into NP [61]. The second approach involves a one-step procedure where the NP are directly generated via different mechanisms such as nanoprecipitation or ionic gelation [86]. An example of each of the two general approaches is summarized in the following.
Ionic gelation, also known as ionotropic gelation or polyelectrolyte complexation involves the gradual addition of a cross-linking agent (tripolyphosphate, glutardehyde etc.) into an aqueous solution of chitosan under continuous stirring to form hydrogels [87]. The polyanions from the cross-linker forms a meshwork of structures by interacting with the polyvalent cations within chitosan, leading to gelation [88]. APIs can be loaded into these hydrogels during the production where it becomes encapsulated or added to the formed NP, where it can be adsorbed into the matrix. The choice of the cross-linker should be matched to the desired physical characteristics of the NP, such as mechanical strength, as well as safety profiles. For example, glutardehyde reported to be toxic when used in high concentrations and results in NP with low mechanical strength. This has been attributed to its double bond (–C=N–) association with the amine group in chitosan [89]. Genipin is a natural cross-linker obtained from iridoid glucoside (geniposide) and present in gardenia fruits that can be cross-linked with chitosan. It displays slower degradation rate than glutaraldehyde and possess higher biocompatibility. Sodium tripolyphosphate (STPP) displays better crosslinker characteristics than each of the above because of its inorganic nature and consequently, results in production of chitosan NP with better mechanical stability. The size dimension derived from STPP gelled chitosan NP is of lower order as well. Another attractive feature of STPP is that it is nontoxic, relatively inexpensive, multivalent, has quick gelling property and thus, widely utilised as a crosslinker in chitosan-based NP [90, 91, 92].
Polymeric nano-emulsions are formulated whereby organic solvent is added to a solution of chitosan with surfactant and mixed via sonication [93]. Basically, the emulsion droplets are converted into NP suspension as the organic solvent evaporates by continuous magnetic stirring at room temperature. The NP suspensions are then centrifuged, washed with distilled water to remove additives such as surfactants and finally lyophilized [94]. Poovi et al. encapsulated the poorly water-soluble drug, repaglinide, into chitosan NP using the emulsion evaporation for sustained release. They proved that the NP exhibited a controlled release of repaglinide and obtained a high drug loading (11.22% w/w) and encapsulation efficiency (97.0%) [95]. In another study, Lee et al. employed solvent evaporation method to formulate polymeric NP from chitosan derivatives fluorescein isothiocyanate (FITC) - conjugated glycol CSs (FGCs) using diluted chloroform as the solvent. Size range of 150–500 nm were obtained and the NP remained stable in phosphate buffered saline for 20 days at 37°C [96].
As mentioned in sections 4.1 and 4.2, extensive research presented the potential of chitosan as an oral absorption enhancer owing to its mucoadhesive properties and ability to loosen tight junctions within the GI epithelia, hence permitting the passage of macromolecular therapeutics across a “well-organised” epithelia [100]. Moreover, due to various characteristics; i.e. non-toxic, biodegradable, biocompatible, antimicrobial property etc. [104], chitosan NP hold promise as a suitable oral delivery vehicle for a wide spectrum of therapeutics including, anti-cancer drugs, antibacterial agents, polyphenolic compounds and protein drugs.
Chemotherapeutic APIs usually exhibit low bioavailability following oral administration. Several studies have investigated chitosan-based NP as a possible delivery system to address this issue. For example, doxorubicin (Dox), broadly employed to treat breast, bladder and other cancers, is typically delivered intravenously. The oral bioavailability of Dox is low due to efflux transporter P-glycoprotein, which identifies Dox as a substrate, restraining its cellular uptake [105]. In 2013, Feng et al. developed chitosan/o-carboxymethyl chitosan (CS/CMCS) NP as a pH responsive carrier for the oral delivery of Dox. They investigated the bioavailability of orally administered Dox-CS/CMCS NP and free Dox drug on Sprague–Dawley rats. Negligible Dox was detected in plasma after the oral dosage of free Dox, representing its poor absorption. On the other hand, 2.3-folds increase in plasma concentration of Dox was registered after an oral dose of Dox-CS/CMCS NP. Moreover, accumulation of Dox in the liver, spleen and lungs were demonstrated in rats treated with oral Dox- CS/CMCS NP, as opposed to DoX solution which was more concentrated in the kidneys. They concluded that the NP matrix improved the intestinal absorption of Dox and thus improved oral bioavailability [106].
Gemcitabine (Gem) is a widely prescribed anticancer agent used in pancreatic, lung and advanced colon cancer. Oral administration of Gem results in low oral bioavailability, high first-pass clearance gastrointestinal toxicity, such as nausea, vomiting and diarrhoea [107]. Hosseinzadeh et al. synthesised and characterised chitosan/Pluronic® F-127 (Gem-Chi/PF) NP in oral delivery of Gem for the treatment of colon cancer.
Chitosan impedes the growth of bacteria, fungi, and yeast [109]. It exhibits potential antimicrobial properties at pH below 6.0 because of the positively charged – NH3+ at the C-2 position within the glucosamine. Low molecular weight chitosan derived NP integrate with bacterial DNA, impeding mRNA synthesis. Conversely, the NH3+ in high molecular chitosan derived NP interact with the negatively-charged cell wall in microorganisms and subsequently amend cell permeability [110]. Alqahtani et al. formulated chitosan NP from high and low molecular weight variants to encapsulate the non-antibiotic diclofenac sodium (DIC). The antibacterial properties of NP from low and high molecular weight of chitosan on
Secondary plant metabolites in the form of polyphenolic compounds have gained wide attention by scientists due to their wide spectrum of pharmacological activities, including antioxidant, antimicrobial and anticancer properties. Most however suffer from poor systemic bioavailability following oral administration due to low solubility and susceptibility to GI degradation. To overcome this constraint, chitosan-based NP have been proposed as a possible delivery intervention, which not only protect these APIs from GI degradation but also improves bioavailability [112]. Curcumin (CUR) is a polyphenol that has been studied extensively. It is derived from the rhizomes of
Proteins are the building blocks of life and required in replicating organisms. Their high molecular weight, chemical and enzymatic susceptibility in the GIT, low diffusion rate through the mucosa barrier and fast systemic clearance, limit their delivery via oral route. As a result, most proteins are administered parenterally. Fortunately, chitosan-based NP are emerging as promising means for the delivery of protein drugs by the oral route through a combination of shielding GI pH, enzymatic degradation and facilitation of epithelial uptake [117]. In a study by He et al., chitosan-STPP insulin NP (CS/STPP/insulin) were orally administered to Type I diabetic rat models in comparison to free insulin solution. Free insulin solution failed to elicit any difference in the blood glucose level, whilst CS/STPP/insulin NP distinctly reduced the blood glucose levels by up to 59% within 8 hours. Crucially, CS/TPP/insulin NP allowed for a fast recovery of blood sugar level when fasting was halted. Moreover, the CS/TPP/insulin NP exhibited negligible toxicity to liver enzymes, confirming the safety profile of the orally delivered CS/TPP/insulin NP. They concluded that CS/TPP NP are an effective oral delivery vehicle for insulin [118]. In another study, Tan et al. demonstrated better
The oral route of administration remains formidable in the systemic delivery of therapeutics. It affords patient compliance, ease of administration and flexibility and remains the favourite choice for administration by patients. However, orally administered therapeutics may undergo premature release in the upper GIT which may render them to enzymatic or pH degradation. Therapeutics that are delivered to the absorptive window are susceptible to efflux pump and metabolic enzymes (e.g., cytochrome P450 enzymes) within the GIT epithelia, which is itself a structural barrier. Scientist involved in the design of therapeutics intended for GI delivery must be cognizant of the above constraints and balance these with the physicochemical properties of the therapeutic. Recent evidence attest to the fact that appropriately formulated NP may be fit for this pursuit. In this regard, chitosan NP is the subject of intense interest because it is readily available, biocompatible, biodegradable, mucoadhesive and influences traversing of therapeutics across the GI epithelia. We expect to see more evidence on the application of chitosan in the oral delivery of therapeutics, especially in the form of NP. Further studies on toxicity related issues
Apomixis provides a method for cloning plants through seeds, so that it is of value for agriculture used to fix hybrid vigor and other hopeful breeding materials that are positioned in the middle breeding process [1, 2]. Apomixis is usually classified into three major mechanisms, apospory, diplospory and adventitious embryogeny [1]. Among the three mechanisms, apospory is considered as the most important one in agriculture because it does not undergo meiosis to propagate through the seed. Bahia grass (
Recently, some challenges using differential interference-contrast microscopy (DIC) technology have been conducted in all organs of plants. For example, in the meiotic chromosome [10], in petal development and ethylene biosynthesis [11], in nucleolus morphological changes [12], in pharmacology and cell biology [13], in xylem differentiation [14], in single microtubules [15]. In particular, near field DIC provides the ability to illuminate two neighboring points on the sample simultaneously, which shows that by modulating the two wavelengths employed in exciting such a probe, phase difference information can be retrieved through measuring the near field photoinduced force at the difference of the two modulation frequencies [16]. And more, two phases (cell structure and fluorescence) that appeared concurrently in the same sample and could be observed in
We choose bahia grass as a monocotyledons species as that would be particularly amenable to a molecular study of apomixis. It is shorter a plant and easily to cultivate among the important forage grasses. Recently, the somatic embryogenesis and plant regeneration system of bahia grass has been established in developing gene introduction techniques [18]. To clarify the molecular process controlling apomixis in
In this study, the major objectives were, using the microscopy method of DIC I) to make it clear the cytological and quantitative observations of AIC appearance and its development in bahia grass, and to estimate the period of AIC appearance using ovary length as an index; and II) to clarify the process of polyembryonic seed set in facultatively apomictic ovules, and to provide information for estimation of the degree of apomixis or sexual of
Four apomictic bahia grass varieties and two obligate sexual varieties were chosen for this study. These materials were kindly provided from Osumi breeding branch, Kagoshima Prefecture Agricultural Experiment station, Japan. For the 2 obligate sexual materials, Nangoku and C 1, the former is a diploid (2n = 20) variety [31], and the latter is a diploid (2n = 20) trace. About four apomictic varieties, they are tetraploid [32].
One hundred to over 300 buds or flowers before and at anthesis were collected for each variety for embryo sac analysis. The buds and flowers were fixed in FPA50 (formalin propionic acid: 50% ethanol = 5: 5: 90) for 5–7 days at 4°C [20, 21, 33, 34]. Ovaries were picked out carefully from fixed buds or flowers under microscope by using needle and tweezers, placed in 70% ethanol followed dehydration series (70, 80, 90, 100% ethanol), and cleared in Herr\'s benzyl-benzoate-four-and-a-half fluid [35] for over 2 h at 0–4°C. The observations were conducted using DIC.
The frequencies of apospory and sexual were calculated at anthesis according to the schematic of Figure 1 [36]. Here, apospory was classified into two types,
Schematic representation of embryo sac development during sexual gametogenesis and the apospory forms in
Until megasporogenesis there showed similar behaviors in both sexual and apospory varieties. After megasporogenesis, however, different events from sexual varieties were observed in apospory varieties. While the formed megaspore became almost unfunctional and degenerated with the membrane disappeared, AIC-derived from enlarged unreduced nucellar cells appeared from a different direction and usually entered the space around the degenerated or surviving megaspore (Figure 2(1), [36]). In Figure 2(2) and (3), there showed coexistence of AIC and degenerated or surviving megaspore.
Aposporous embryo sac initial cell (AIC) appearance and AIC-derived embryo sac maturity in apomictic bahia grass (
In general, the AIC forms the embryo sac through a special process. The AIC undergoes mitosis two times and forms 2-, 4-nucleate. And no antipodal was found (Figure 2(4)). The cell division only occurred in the half-space of the embryo sac in the micropylar end. It is different from the sexual one which usually occupied the whole space of the sac. And then, the 4 nucleates developed to complete their parts in order (Figure 2(5)). In common, a mature embryo sac formed with one egg cell, one synergid and two polar nuclei (Figure 2(6)). In some rare cases,
In most ovules of apomictic plants, AICs appeared continuously as the ovary length increased. The numbers of AICs were increased between the formations of functional megaspore and mature 4- or 8-nucleates according to the ovary length. For example, the ovary length in "competitor " was ca 520 μm when the first AIC appeared, and ca 496 μm when the third one did. However, 5 AICs appeared between 520 μm to 624 μm (688 μm) in ovary length. So were the other varieties. These values of ovary length indicated that AICs in the same ovule did not seem to differentiate synchronously.
From the length of ovaries, the stages of the ovary containing degenerated embryo sac were from 4-nucleate to their embryo sac maturity in apomictic plants, and from 4- and 8-nucleate to embryo sac maturity in sexual plants, respectively. On the other hand, the stage of functional megaspore showed a range of ovary length so wide it became very close to the value of degenerated ovaries.
Ten types of embryo sacs were observed and most of the embryo sacs belonged to PS type (Table 1, [36]). In apomicts, most ovules showed the number of embryo sacs more than one. And the ovules with different embryo sacs (5PS to 2PS, PS +PNn to 4PS +PNn, S, PNn) were observed. The percentage of S type (
Varieties | No. ovules observed | No. sterile ovules | Types of embryo sacs1 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
5PS | 4PS | 3PS | 2PS | S | PS+PNn | 2PS+PNn | 3PS+PNn | 4PS+PNn | PNn | |||
Competitor | 60 | 1 | 4 | 7 | 14 | 14 | 8 | 4 | 4 | 1 | 2 | 1 |
Nanou | 35 | 2 | 1 | 2 | 7 | 11 | 3 | 3 | 2 | 1 | 3 | |
Tifron | 44 | 14 | 2 | 1 | 4 | 13 | 4 | 2 | 2 | 1 | 1 | |
Common | 43 | 6 | 3 | 5 | 10 | 13 | 2 | 3 | 1 |
Number of ovules with different types of embryo sacs at anthesis in apomictic varieties of
PS:
Among the 4 varieties, the frequencies of PN type were from 5.0% to 30.3%, and that of PS type were from 60.7% to 90.0%. Total frequencies of apospory were 86.4%, 91.1%, 87.1% and 95.0% in competitor, Nanou, Tifton and Common, respectively.
There are no differences observed between obligate sexual and apomictic plants until megasporogenesis in bahia grass. After megasporogenesis, however, different events are followed in embryo sac formation. Sexual ovules proceeded in a manner typical of the Gramineae family, i.e., functional megaspores divided and formed a mature 8- nucleate embryo sac as reported in
The earliest AIC that appeared in ovule always located in micropylar end, as the ovary grows, the later appeared are located along with the first AIC and being apart from it. To understand the mechanism of AIC appearance, we selected ovary length as an index and measured the ovaries when they were observed in different AIC appearance. From the range of ovary length, AICs do not appear together in same time. Instead, they seemed following a continuous course and appeared one by one during the period from megasporogenesis even to the first embryo sac maturity. According to the ovary length compared with the morphology of spikes, AICs appeared in the period of spike emerging to open at anthesis. With regard to the ovary length measured we could collect every stage of embryo sac to apply apomixis gene cloning. Sterile ovules with degenerated embryo sac appeared in both sexual and apomictic varieties based on the observation and quantitative analysis of ovary length. The ovary length of the ovary in which the first AIC appeared was longer than that of ovary the functional megaspore appeared in all varieties, indicating that the aposporous phenomenon of AIC appearance is initiated after megasporogenesis. Further, the ovary length of the ovary staged in functional megaspore was wide and close to the ovary lengths of the ovaries showing degeneration of megaspores in different developed stages. These results indicated indirectly that the development of sexual embryo sac derived from megaspore is often terminated accompanied by AIC appearance in many aposporous apomicts around the stage of megasporogenesis [41, 42]. Which one of megaspore or AIC firstly showed the signal to terminate or to appear will be interesting to further researches of apomixis. In the present study, 10 types of embryo sac formation were observed in
At 0 DAA, most of the embryo sac located in micropylar end became typical ones. In the accessions tested, the percentage of the typical embryo sacs in micropylar end was 90.9–100% higher than that in the other end (40.4–86.0%) (Table 2, [43]). If the typical one is S type, it consists of 8-nucleates of egg cell, 2 synergids and 3 antipodal. If the typical one is PN or PS type, it consists of 4-nucleates of egg cell, synergid and 2 polar nuclei.
Varieties | No. ovules observed (A) | No. embryo sacs (B) | Mean no. (B/A) | No. embryo sacs | |
---|---|---|---|---|---|
Micropylar | Others | ||||
Competidor | 59 | 171 | 2.9 | 59 (100%) | 112 (78.6%) |
Nanou | 33 | 85 | 2.6 | 30 (90.9%) | 52 (40.4%) |
Tifton | 31 | 51 | 1.6 | 31 (96.8%) | 20 (80.0%) |
Common | 40 | 90 | 2.3 | 40 (92.5%) | 50 (86.0%) |
Distribution of matured embryo sacs at anthesis in facultatively apomictic varieties of
At 6 h after anthesis, polar nuclei firstly started to divide to mean 9.0 cells in 55 of 59 ovules of Competitor. At 1 DAA, first division of egg cell was observed while the endosperm had reached mean 38.8 cells. This phenomenon is same to
In polyembryonic ovules, the embryo sac in micropylar end developed dominantly when compared with that in the other ends. Figure 3 showed the developments of both embryo and endosperm of the sac in micropylar end, and the other sacs were crowded out to chalazal end, with dividing egg embryo and polar nuclei [36].
Multiple embryo formation in the same embryo sac in
The ovules containing dominantly developed embryo sacs were investigated at 4 DAA in 4 varieties. The results showed in Table 3 indicated that, 56–87% of the ovules contained developed sacs in micropylar end in all the accessions, and in contrast, on other ends, 0% in 3 varieties and 3% in one variety.
Varieties | No. ovules observed A | No. embryo sacs B | ES in miciopylar end | ES in the other end | ||||
---|---|---|---|---|---|---|---|---|
embryo and endosperm C(C/A) | embryo only | endosperm only | embryo and endosperm D(D/A) | embryo only E(E/B) | endosperm only | |||
Competitor | 30 | 41 | 26 (87%) | 0 | 4 | 0 (0%) | 7 (17%) | 0 |
Nanou | 27 | 28 | 15 (56%) | 1 | 0 | 0 (0%) | 1 (4%) | 0 |
Tifton | 30 | 35 | 21 (66%) | 0 | 2 | 0 (0%) | 3 (9%) | 0 |
Common | 30 | 38 | 23 (77%) | 0 | 0 | 1 (3%) | 5 (13%) | 0 |
Development of embryo sac (ES) in the flowers 4 d after anthesis in facultatively apomictic varieties of
Some other cases appeared different from the above. 1) A single embryo sac in micropylar end contains a well-developed embryo and unfertilized 2 nuclei. 2) A single embryo sac contains 2 embryos and well-developed endosperm. 3) The embryo sac degenerated in micropylar end, and in chalazal end, the sac with developed embryo and unfertilized 2 nuclei. And the data of other types could be known from Table 3. Among the 4 varieties, the number of the embryo sac in micropylar end containing only embryo was 1 of 27 ovules in Nanou, and the numbers containing only endosperm were 4 of 30 ovules in Competitor, 2 of 30 ovules in Tifton, respectively. In contrast, in the other end, the numbers of the sacs containing only embryo were 4–17% in all the varieties, and the number containing only endosperm was 0% in the varieties.
At 6 DAA, the developed embryo and endosperm of the sac in micropylar end occupied the embryo sac it located in, and at the same time, the space of the whole ovules was almost occupied by the developed sac (Figure 3a). The embryo usually exists with near globular shape, and it is surrounded by well-developed endosperm. For the embryo sacs in the other ends, they usually were squeezed out to outside of the developed sac. However, those sacs showed continuous development. Some egg cells divided well, and formed embryos usually located in opposite side to micropylar end (Figure 3a), or the neighbor (Figure 3b and
In emasculated ovules observed at 4 DAA, no developed embryo sac distinguished in 60 ovules of 4 varieties. The parthenogenesis rate was 0% in all the 4 varieties. At 15 DAA, the inflorescences that were emasculated and then isolated from any pollen source failed to produce seed. So, pseudogamous is essential for seed set in
In facultative apomictic bahia grass, AICs appeared one by one, and then, they became multiple embryo sacs in same ovule, as the ovary length increased [36]. And as the first AIC usually appeared and located in the micropylar end, 92.5 to 100% of embryo sacs closest to micropylar end of ovule matured at anthesis observed in this study (Table 2). On the other hand, the embryo sacs located in other end showed 40.4 to 86.0% mature rates at anthesis. The AIC appearance age (order) maybe influence the mature of apomictic embryo sacs themselves. It could be considered that the first appeared AIC in micropylar end has the temporal dominant in formation and maturity of the embryo sac when compared with the other sacs. And for the fertilization chance, the sac has also the positional dominant, as it was closest to synergid cell through which pollen tube penetrates and finishes fertilization. Therefore, the sac derived from first AIC located in micropylar end has the advantage of fertilization. On the observations of ovules at 4 DAA, the rates of developed embryo sacs with embryo and endosperm were from 56 to 87% in the sacs of micropylar end (Table 3). On the other hand, the other sacs were 0% in 3 varieties, and one variety was 3%. Therefore, the sac in micropylar end has the advantage of seed set. This result also supported the hypothesis that the embryo of developed sac in micropylar end, in final, became a seed-forming embryo [20]. Using this method, we can estimate the degree of apomixis or sexual of any facultative apomictic materials used, based on the analysis of embryo sacs in micropylar end at anthesis.
Different events were observed on the seed set between guinea grass and bahia grass. In guinea grass, the other embryo sacs were crowed out to the chalazal end by the developed micropylar sac, and in final, they were completely degenerated after 10 DAA [21]. In contrast, the rates of embryos formed in the other embryo sacs were 4–17% in 4 accessions of bahia grass used in this study. These are higher than that (0%) in 5 accessions and that (2%) in one accession of guinea grass [21]. This evident was also observed from embryo sac analysis (Figure 3). As the sac derived from AIC contains 2n level reproductive cells, egg cell does not need fertilization. However, for the endosperm formation, fertilization between central cell and sperm cell is needed. And egg cell usually starts division followed the endosperm cell formation. The other sacs also follow the same manner. As the embryo sac developed advantageously in micropylar end, it could be considered that the egg cells in the other sacs were developed vigorously in different places of ovules. In that case, only embryo formed but no endosperm. In the polyembryonic ovules, the embryos located in the other sacs usually presented close to the well-developed endosperm of the micropylar sac. The 2 kinds of embryos in the same ovule seemed sharing the endosperm of the micropylar sac. Maybe that is why the embryos in different embryo sacs could coexist in the same ovule. At the germination experiment, twines, or multiple seedlings (>5%) were observed (data not shown). This means the different embryos have the same germination capacity. For the endosperm balance number, some reporters have discussed the requirement for balance between maternal and paternal contributions to the endosperm formation [44, 45, 46, 47, 48]. When they used different ploidies and sexual materials, endosperm balance in terms of maternal to paternal ratio, 2:1 was considered balanced and should produce normal endosperm. And they indicated that unreduced embryo sacs with one central-cell nucleus (4 factors), when fertilized by a sperm (2 factors), would result in the proper endosperm ratio of 2m:1p, and the closed percentages were obtained between embryo sacs with a single central-cell nucleus and ovaries with endosperm developing 6 and 8 d after pollination. The result of aposporous guinea grass reported by Chen and Kozono [21] also supported the above explanation. In grasses, the most common type of unreduced embryo sac is the 4-nucleates
Recently, the mechanisms of reproductive characterization [49], molecular and genetic regulation [50, 51, 52], and its utilization [53, 54] have been discovered consequently in
In this study, using the plant materials of several apomictic and obligately sexual varieties of bahia grass (
This study was partly supported by the Grant-in-Aid for Scientific Research (C) of the Ministry of Education, Culture, Science and Sports of Japan, No. 10660010.
Authors have declared that no competing interests exist.
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However, agricultural malpractices like excessive use of chemical fertilizers and pesticides, as well as climate change have aggravated the effects of biotic and abiotic stresses on crop productivity. These led to the degradation of ecosystem, leaving bad impacts on the soil qualities and water body environment. As an alternative to the rising agricultural energy, the use of Vesicular– Arbuscular Mycorrhizae (AM) may be a better option. Being natural root symbionts, AM provide essential inorganic nutrients to host plants, thereby improving its growth and yield even under stressed conditions. AM fungi can also potentially strengthen the adaptability of a plant to the changing environment, as a bio-fertilizer. The chapter provides a comprehensive up-to-date knowledge on AM fungi as a tool for sustainable agricultural system. 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Lentil crops are vulnerable to a number of diseases caused by fungi, viruses, nematodes, insect pests, parasitic plants and abiotic stresses. Among them, the most significant and serious soil-borne disease is Fusarium wilt (Fusarium oxysporum f.sp. lentis: Fol). Fusarium wilt causes yield loss up to 50% in farmers’ fields. The pathogen showed high levels of phenotypic and genotypic diversity in India, Algeria, Syria and Iran. The disease thrives at 22–25°C temperature and affect lentil either at seedling and vegetative or the reproductive stages of the crop. To minimize yield losses, an integrated management strategy comprising resistant/partial resistant cultivars, adjusting sowing time, bio-control and chemical seed treatments is the best approach to reduce the incidence of the Fusarium wilt of lentil. 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He is also a faculty member in the Molecular Oncology Program. He obtained his MSc and Ph.D. at Oregon State University and Texas Tech University, respectively. He pursued his postdoctoral studies at Rutgers University Medical School and the National Institutes of Health (NIH/NIDDK), USA. His research focuses on biochemistry, biophysics, genetics, molecular biology, and molecular medicine with specialization in the fields of drug design, protein structure-function, protein folding, prions, microRNA, pseudogenes, molecular cancer, epigenetics, metabolites, proteomics, genomics, protein expression, and characterization by spectroscopic and calorimetric methods.",institutionString:"University of Health Sciences",institution:null},{id:"180528",title:"Dr.",name:"Hiroyuki",middleName:null,surname:"Kagechika",slug:"hiroyuki-kagechika",fullName:"Hiroyuki Kagechika",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/180528/images/system/180528.jpg",biography:"Hiroyuki Kagechika received his bachelor’s degree and Ph.D. in Pharmaceutical Sciences from the University of Tokyo, Japan, where he served as an associate professor until 2004. He is currently a professor at the Institute of Biomaterials and Bioengineering (IBB), Tokyo Medical and Dental University (TMDU). From 2010 to 2012, he was the dean of the Graduate School of Biomedical Science. Since 2012, he has served as the vice dean of the Graduate School of Medical and Dental Sciences. He has been the director of the IBB since 2020. Dr. Kagechika’s major research interests are the medicinal chemistry of retinoids, vitamins D/K, and nuclear receptors. 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Currently, he is a Professor at Xiangya Hospital of Central South University in China, Fellow of Royal Society of Medicine (FRSM), the European EPMA National Representative in China, Regular Member of American Association for the Advancement of Science (AAAS), European Cooperation of Science and Technology (e-COST) grant evaluator, Associate Editors of BMC Genomics, BMC Medical Genomics, EPMA Journal, and Frontiers in Endocrinology, Executive Editor-in-Chief of Med One. He has\npublished 116 peer-reviewed research articles, 16 book chapters, 2 books, and 2 US patents. 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She is currently leading project entitled Mesenchymal stem cells-the keepers of tissue endogenous regenerative capacity facing up to aging of the musculoskeletal system funded by Slovenian Research Agency.",institutionString:null,institution:{name:"University of Ljubljana",country:{name:"Slovenia"}}},{id:"357453",title:"Dr.",name:"Radheshyam",middleName:null,surname:"Maurya",slug:"radheshyam-maurya",fullName:"Radheshyam Maurya",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/357453/images/16535_n.jpg",biography:null,institutionString:null,institution:{name:"University of Hyderabad",country:{name:"India"}}},{id:"418340",title:"Dr.",name:"Jyotirmoi",middleName:null,surname:"Aich",slug:"jyotirmoi-aich",fullName:"Jyotirmoi Aich",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000038Ugi5QAC/Profile_Picture_2022-04-15T07:48:28.png",biography:"Biotechnologist with 15 years of research including 6 years of teaching experience. Demonstrated record of scientific achievements through consistent publication record (H index = 13, with 874 citations) in high impact journals such as Nature Communications, Oncotarget, Annals of Oncology, PNAS, and AJRCCM, etc. Strong research professional with a post-doctorate from ACTREC where I gained experimental oncology experience in clinical settings and a doctorate from IGIB where I gained expertise in asthma pathophysiology. A well-trained biotechnologist with diverse experience on the bench across different research themes ranging from asthma to cancer and other infectious diseases. An individual with a strong commitment and innovative mindset. Have the ability to work on diverse projects such as regenerative and molecular medicine with an overall mindset of improving healthcare.",institutionString:"DY Patil Deemed to Be University",institution:null},{id:"349288",title:"Prof.",name:"Soumya",middleName:null,surname:"Basu",slug:"soumya-basu",fullName:"Soumya Basu",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000035QxIDQA0/Profile_Picture_2022-04-15T07:47:01.jpg",biography:"Soumya Basu, Ph.D., is currently working as an Associate Professor at Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pune, Maharashtra, India. With 16+ years of trans-disciplinary research experience in Drug Design, development, and pre-clinical validation; 20+ research article publications in journals of repute, 9+ years of teaching experience, trained with cross-disciplinary education, Dr. Basu is a life-long learner and always thrives for new challenges.\r\nHer research area is the design and synthesis of small molecule partial agonists of PPAR-γ in lung cancer. She is also using artificial intelligence and deep learning methods to understand the exosomal miRNA’s role in cancer metastasis. Dr. Basu is the recipient of many awards including the Early Career Research Award from the Department of Science and Technology, Govt. of India. She is a reviewer of many journals like Molecular Biology Reports, Frontiers in Oncology, RSC Advances, PLOS ONE, Journal of Biomolecular Structure & Dynamics, Journal of Molecular Graphics and Modelling, etc. She has edited and authored/co-authored 21 journal papers, 3 book chapters, and 15 abstracts. She is a Board of Studies member at her university. She is a life member of 'The Cytometry Society”-in India and 'All India Cell Biology Society”- in India.",institutionString:"Dr. D.Y. Patil Vidyapeeth, Pune",institution:{name:"Dr. D.Y. Patil Vidyapeeth, Pune",country:{name:"India"}}},{id:"354817",title:"Dr.",name:"Anubhab",middleName:null,surname:"Mukherjee",slug:"anubhab-mukherjee",fullName:"Anubhab Mukherjee",position:null,profilePictureURL:"https://intech-files.s3.amazonaws.com/0033Y0000365PbRQAU/ProfilePicture%202022-04-15%2005%3A11%3A18.480",biography:"A former member of Laboratory of Nanomedicine, Brigham and Women’s Hospital, Harvard University, Boston, USA, Dr. Anubhab Mukherjee is an ardent votary of science who strives to make an impact in the lives of those afflicted with cancer and other chronic/acute ailments. He completed his Ph.D. from CSIR-Indian Institute of Chemical Technology, Hyderabad, India, having been skilled with RNAi, liposomal drug delivery, preclinical cell and animal studies. He pursued post-doctoral research at College of Pharmacy, Health Science Center, Texas A & M University and was involved in another postdoctoral research at Department of Translational Neurosciences and Neurotherapeutics, John Wayne Cancer Institute, Santa Monica, California. In 2015, he worked in Harvard-MIT Health Sciences & Technology as a visiting scientist. He has substantial experience in nanotechnology-based formulation development and successfully served various Indian organizations to develop pharmaceuticals and nutraceutical products. He is an inventor in many US patents and an author in many peer-reviewed articles, book chapters and books published in various media of international repute. Dr. Mukherjee is currently serving as Principal Scientist, R&D at Esperer Onco Nutrition (EON) Pvt. Ltd. and heads the Hyderabad R&D center of the organization.",institutionString:"Esperer Onco Nutrition Pvt Ltd.",institution:null},{id:"319365",title:"Assistant Prof.",name:"Manash K.",middleName:null,surname:"Paul",slug:"manash-k.-paul",fullName:"Manash K. Paul",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/319365/images/system/319365.png",biography:"Manash K. Paul is a Principal Investigator and Scientist at the University of California Los Angeles. He has contributed significantly to the fields of stem cell biology, regenerative medicine, and lung cancer. His research focuses on various signaling processes involved in maintaining stem cell homeostasis during the injury-repair process, deciphering lung stem cell niche, pulmonary disease modeling, immuno-oncology, and drug discovery. He is currently investigating the role of extracellular vesicles in premalignant lung cell migration and detecting the metastatic phenotype of lung cancer via machine-learning-based analyses of exosomal signatures. Dr. Paul has published in more than fifty peer-reviewed international journals and is highly cited. He is the recipient of many awards, including the UCLA Vice Chancellor’s award, a senior member of the Institute of Electrical and Electronics Engineers (IEEE), and an editorial board member for several international journals.",institutionString:"University of California Los Angeles",institution:{name:"University of California Los Angeles",country:{name:"United States of America"}}},{id:"311457",title:"Dr.",name:"Júlia",middleName:null,surname:"Scherer Santos",slug:"julia-scherer-santos",fullName:"Júlia Scherer Santos",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/311457/images/system/311457.jpg",biography:"Dr. Júlia Scherer Santos works in the areas of cosmetology, nanotechnology, pharmaceutical technology, beauty, and aesthetics. Dr. Santos also has experience as a professor of graduate courses. Graduated in Pharmacy, specialization in Cosmetology and Cosmeceuticals applied to aesthetics, specialization in Aesthetic and Cosmetic Health, and a doctorate in Pharmaceutical Nanotechnology. Teaching experience in Pharmacy and Aesthetics and Cosmetics courses. She works mainly on the following subjects: nanotechnology, cosmetology, pharmaceutical technology, aesthetics.",institutionString:"Universidade Federal de Juiz de Fora",institution:{name:"Universidade Federal de Juiz de Fora",country:{name:"Brazil"}}},{id:"219081",title:"Dr.",name:"Abdulsamed",middleName:null,surname:"Kükürt",slug:"abdulsamed-kukurt",fullName:"Abdulsamed Kükürt",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/219081/images/system/219081.png",biography:"Dr. Kükürt graduated from Uludağ University in Turkey. He started his academic career as a Research Assistant in the Department of Biochemistry at Kafkas University. In 2019, he completed his Ph.D. program in the Department of Biochemistry at the Institute of Health Sciences. He is currently working at the Department of Biochemistry, Kafkas University. He has 27 published research articles in academic journals, 11 book chapters, and 37 papers. He took part in 10 academic projects. He served as a reviewer for many articles. He still serves as a member of the review board in many academic journals. He is currently working on the protective activity of phenolic compounds in disorders associated with oxidative stress and inflammation.",institutionString:null,institution:{name:"Kafkas University",country:{name:"Turkey"}}},{id:"178366",title:"Dr.",name:"Volkan",middleName:null,surname:"Gelen",slug:"volkan-gelen",fullName:"Volkan Gelen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/178366/images/system/178366.jpg",biography:"Volkan Gelen is a Physiology specialist who received his veterinary degree from Kafkas University in 2011. Between 2011-2015, he worked as an assistant at Atatürk University, Faculty of Veterinary Medicine, Department of Physiology. In 2016, he joined Kafkas University, Faculty of Veterinary Medicine, Department of Physiology as an assistant professor. Dr. Gelen has been engaged in various academic activities at Kafkas University since 2016. There he completed 5 projects and has 3 ongoing projects. He has 60 articles published in scientific journals and 20 poster presentations in scientific congresses. His research interests include physiology, endocrine system, cancer, diabetes, cardiovascular system diseases, and isolated organ bath system studies.",institutionString:"Kafkas University",institution:{name:"Kafkas University",country:{name:"Turkey"}}},{id:"418963",title:"Dr.",name:"Augustine Ododo",middleName:"Augustine",surname:"Osagie",slug:"augustine-ododo-osagie",fullName:"Augustine Ododo Osagie",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/418963/images/16900_n.jpg",biography:"Born into the family of Osagie, a prince of the Benin Kingdom. I am currently an academic in the Department of Medical Biochemistry, University of Benin. Part of the duties are to teach undergraduate students and conduct academic research.",institutionString:null,institution:{name:"University of Benin",country:{name:"Nigeria"}}},{id:"192992",title:"Prof.",name:"Shagufta",middleName:null,surname:"Perveen",slug:"shagufta-perveen",fullName:"Shagufta Perveen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/192992/images/system/192992.png",biography:"Prof. Shagufta Perveen is a Distinguish Professor in the Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia. Dr. Perveen has acted as the principal investigator of major research projects funded by the research unit of King Saud University. She has more than ninety original research papers in peer-reviewed journals of international repute to her credit. She is a fellow member of the Royal Society of Chemistry UK and the American Chemical Society of the United States.",institutionString:"King Saud University",institution:{name:"King Saud University",country:{name:"Saudi Arabia"}}},{id:"49848",title:"Dr.",name:"Wen-Long",middleName:null,surname:"Hu",slug:"wen-long-hu",fullName:"Wen-Long Hu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/49848/images/system/49848.jpg",biography:"Wen-Long Hu is Chief of the Division of Acupuncture, Department of Chinese Medicine at Kaohsiung Chang Gung Memorial Hospital, as well as an adjunct associate professor at Fooyin University and Kaohsiung Medical University. Wen-Long is President of Taiwan Traditional Chinese Medicine Medical Association. He has 28 years of experience in clinical practice in laser acupuncture therapy and 34 years in acupuncture. He is an invited speaker for lectures and workshops in laser acupuncture at many symposiums held by medical associations. He owns the patent for herbal preparation and producing, and for the supercritical fluid-treated needle. Dr. Hu has published three books, 12 book chapters, and more than 30 papers in reputed journals, besides serving as an editorial board member of repute.",institutionString:"Kaohsiung Chang Gung Memorial Hospital",institution:{name:"Kaohsiung Chang Gung Memorial Hospital",country:{name:"Taiwan"}}},{id:"298472",title:"Prof.",name:"Andrey V.",middleName:null,surname:"Grechko",slug:"andrey-v.-grechko",fullName:"Andrey V. Grechko",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/298472/images/system/298472.png",biography:"Andrey Vyacheslavovich Grechko, Ph.D., Professor, is a Corresponding Member of the Russian Academy of Sciences. He graduated from the Semashko Moscow Medical Institute (Semashko National Research Institute of Public Health) with a degree in Medicine (1998), the Clinical Department of Dermatovenerology (2000), and received a second higher education in Psychology (2009). Professor A.V. Grechko held the position of Сhief Physician of the Central Clinical Hospital in Moscow. He worked as a professor at the faculty and was engaged in scientific research at the Medical University. Starting in 2013, he has been the initiator of the creation of the Federal Scientific and Clinical Center for Intensive Care and Rehabilitology, Moscow, Russian Federation, where he also serves as Director since 2015. He has many years of experience in research and teaching in various fields of medicine, is an author/co-author of more than 200 scientific publications, 13 patents, 15 medical books/chapters, including Chapter in Book «Metabolomics», IntechOpen, 2020 «Metabolomic Discovery of Microbiota Dysfunction as the Cause of Pathology».",institutionString:"Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology",institution:null},{id:"199461",title:"Prof.",name:"Natalia V.",middleName:null,surname:"Beloborodova",slug:"natalia-v.-beloborodova",fullName:"Natalia V. Beloborodova",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/199461/images/system/199461.jpg",biography:'Natalia Vladimirovna Beloborodova was educated at the Pirogov Russian National Research Medical University, with a degree in pediatrics in 1980, a Ph.D. in 1987, and a specialization in Clinical Microbiology from First Moscow State Medical University in 2004. She has been a Professor since 1996. Currently, she is the Head of the Laboratory of Metabolism, a division of the Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Moscow, Russian Federation. N.V. Beloborodova has many years of clinical experience in the field of intensive care and surgery. She studies infectious complications and sepsis. She initiated a series of interdisciplinary clinical and experimental studies based on the concept of integrating human metabolism and its microbiota. Her scientific achievements are widely known: she is the recipient of the Marie E. Coates Award \\"Best lecturer-scientist\\" Gustafsson Fund, Karolinska Institutes, Stockholm, Sweden, and the International Sepsis Forum Award, Pasteur Institute, Paris, France (2014), etc. Professor N.V. Beloborodova wrote 210 papers, five books, 10 chapters and has edited four books.',institutionString:"Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology",institution:null},{id:"354260",title:"Ph.D.",name:"Tércio Elyan",middleName:"Azevedo",surname:"Azevedo Martins",slug:"tercio-elyan-azevedo-martins",fullName:"Tércio Elyan Azevedo Martins",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/354260/images/16241_n.jpg",biography:"Graduated in Pharmacy from the Federal University of Ceará with the modality in Industrial Pharmacy, Specialist in Production and Control of Medicines from the University of São Paulo (USP), Master in Pharmaceuticals and Medicines from the University of São Paulo (USP) and Doctor of Science in the program of Pharmaceuticals and Medicines by the University of São Paulo. Professor at Universidade Paulista (UNIP) in the areas of chemistry, cosmetology and trichology. Assistant Coordinator of the Higher Course in Aesthetic and Cosmetic Technology at Universidade Paulista Campus Chácara Santo Antônio. Experience in the Pharmacy area, with emphasis on Pharmacotechnics, Pharmaceutical Technology, Research and Development of Cosmetics, acting mainly on topics such as cosmetology, antioxidant activity, aesthetics, photoprotection, cyclodextrin and thermal analysis.",institutionString:null,institution:{name:"University of Sao Paulo",country:{name:"Brazil"}}},{id:"334285",title:"Ph.D. Student",name:"Sameer",middleName:"Kumar",surname:"Jagirdar",slug:"sameer-jagirdar",fullName:"Sameer Jagirdar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/334285/images/14691_n.jpg",biography:"I\\'m a graduate student at the center for biosystems science and engineering at the Indian Institute of Science, Bangalore, India. I am interested in studying host-pathogen interactions at the biomaterial interface.",institutionString:null,institution:{name:"Indian Institute of Science Bangalore",country:{name:"India"}}},{id:"329248",title:"Dr.",name:"Md. Faheem",middleName:null,surname:"Haider",slug:"md.-faheem-haider",fullName:"Md. Faheem Haider",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329248/images/system/329248.jpg",biography:"Dr. Md. Faheem Haider completed his BPharm in 2012 at Integral University, Lucknow, India. In 2014, he completed his MPharm with specialization in Pharmaceutics at Babasaheb Bhimrao Ambedkar University, Lucknow, India. He received his Ph.D. degree from Jamia Hamdard University, New Delhi, India, in 2018. He was selected for the GPAT six times and his best All India Rank was 34. Currently, he is an assistant professor at Integral University. Previously he was an assistant professor at IIMT University, Meerut, India. He has experience teaching DPharm, Pharm.D, BPharm, and MPharm students. He has more than five publications in reputed journals to his credit. Dr. Faheem’s research area is the development and characterization of nanoformulation for the delivery of drugs to various organs.",institutionString:"Integral University",institution:{name:"Integral University",country:{name:"India"}}},{id:"329795",title:"Dr.",name:"Mohd Aftab",middleName:"Aftab",surname:"Siddiqui",slug:"mohd-aftab-siddiqui",fullName:"Mohd Aftab Siddiqui",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329795/images/system/329795.png",biography:"Dr. Mohd Aftab Siddiqui is an assistant professor in the Faculty of Pharmacy, Integral University, Lucknow, India, where he obtained a Ph.D. in Pharmacology in 2020. He also obtained a BPharm and MPharm from the same university in 2013 and 2015, respectively. His area of research is the pharmacological screening of herbal drugs/natural products in liver cancer and cardiac diseases. He is a member of many professional bodies and has guided many MPharm and PharmD research projects. Dr. Siddiqui has many national and international publications and one German patent to his credit.",institutionString:"Integral University",institution:null},{id:"255360",title:"Dr.",name:"Usama",middleName:null,surname:"Ahmad",slug:"usama-ahmad",fullName:"Usama Ahmad",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/255360/images/system/255360.png",biography:"Dr. Usama Ahmad holds a specialization in Pharmaceutics from Amity University, Lucknow, India. He received his Ph.D. from Integral University, Lucknow, India, with his work titled ‘Development and evaluation of silymarin nanoformulation for hepatic carcinoma’. Currently, he is an Assistant Professor of Pharmaceutics, at the Faculty of Pharmacy, Integral University. He has been teaching PharmD, BPharm, and MPharm students and conducting research in the novel drug delivery domain. From 2013 to 2014 he worked on a research project funded by SERB-DST, Government of India. He has a rich publication record with more than twenty-four original journal articles, two edited books, four book chapters, and several scientific articles to his credit. He is a member of the American Association for Cancer Research, the International Association for the Study of Lung Cancer, and the British Society for Nanomedicine. Dr. Ahmad’s research focus is on the development of nanoformulations to facilitate the delivery of drugs.",institutionString:"Integral University",institution:{name:"Integral University",country:{name:"India"}}},{id:"333824",title:"Dr.",name:"Ahmad Farouk",middleName:null,surname:"Musa",slug:"ahmad-farouk-musa",fullName:"Ahmad Farouk Musa",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/333824/images/22684_n.jpg",biography:"Dato’ Dr Ahmad Farouk Musa\nMD, MMED (Surgery) (Mal), Fellowship in Cardiothoracic Surgery (Monash Health, Aust), Graduate Certificate in Higher Education (Aust), Academy of Medicine (Mal)\n\n\n\nDato’ Dr Ahmad Farouk Musa obtained his Doctor of Medicine from USM in 1992. He then obtained his Master of Medicine in Surgery from the same university in the year 2000 before subspecialising in Cardiothoracic Surgery at Institut Jantung Negara (IJN), Kuala Lumpur from 2002 until 2005. He then completed his Fellowship in Cardiothoracic Surgery at Monash Health, Melbourne, Australia in 2008. He has served in the Malaysian army as a Medical Officer with the rank of Captain upon completing his Internship before joining USM as a trainee lecturer. He is now serving as an academic and researcher at Monash University Malaysia. He is a life-member of the Malaysian Association of Thoracic & Cardiovascular Surgery (MATCVS) and a committee member of the MATCVS Database. He is also a life-member of the College of Surgeons, Academy of Medicine of Malaysia; a life-member of Malaysian Medical Association (MMA), and a life-member of Islamic Medical Association of Malaysia (IMAM). Recently he was appointed as an Interim Chairperson of Examination & Assessment Subcommittee of the UiTM-IJN Cardiothoracic Surgery Postgraduate Program. As an academic, he has published numerous research papers and book chapters. He has also been appointed to review many scientific manuscripts by established journals such as the British Medical Journal (BMJ). He has presented his research works at numerous local and international conferences such as the European Association for Cardiothoracic Surgery (EACTS) and the European Society of Cardiovascular Surgery (ESCVS), to name a few. He has also won many awards for his research presentations at meetings and conferences like the prestigious International Invention, Innovation & Technology Exhibition (ITEX); Design, Research and Innovation Exhibition, the National Conference on Medical Sciences and the Annual Scientific Meetings of the Malaysian Association for Thoracic and Cardiovascular Surgery. He was awarded the Darjah Setia Pangkuan Negeri (DSPN) by the Governor of Penang in July, 2015.",institutionString:null,institution:{name:"Monash University Malaysia",country:{name:"Malaysia"}}},{id:"30568",title:"Prof.",name:"Madhu",middleName:null,surname:"Khullar",slug:"madhu-khullar",fullName:"Madhu Khullar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/30568/images/system/30568.jpg",biography:"Dr. Madhu Khullar is a Professor of Experimental Medicine and Biotechnology at the Post Graduate Institute of Medical Education and Research, Chandigarh, India. She completed her Post Doctorate in hypertension research at the Henry Ford Hospital, Detroit, USA in 1985. She is an editor and reviewer of several international journals, and a fellow and member of several cardiovascular research societies. Dr. Khullar has a keen research interest in genetics of hypertension, and is currently studying pharmacogenetics of hypertension.",institutionString:"Post Graduate Institute of Medical Education and Research",institution:{name:"Post Graduate Institute of Medical Education and Research",country:{name:"India"}}},{id:"223233",title:"Prof.",name:"Xianquan",middleName:null,surname:"Zhan",slug:"xianquan-zhan",fullName:"Xianquan Zhan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/223233/images/system/223233.png",biography:"Xianquan Zhan received his MD and Ph.D. in Preventive Medicine at West China University of Medical Sciences. He received his post-doctoral training in oncology and cancer proteomics at the Central South University, China, and the University of Tennessee Health Science Center (UTHSC), USA. He worked at UTHSC and the Cleveland Clinic in 2001–2012 and achieved the rank of associate professor at UTHSC. Currently, he is a full professor at Central South University and Shandong First Medical University, and an advisor to MS/PhD students and postdoctoral fellows. He is also a fellow of the Royal Society of Medicine and European Association for Predictive Preventive Personalized Medicine (EPMA), a national representative of EPMA, and a member of the American Society of Clinical Oncology (ASCO) and the American Association for the Advancement of Sciences (AAAS). He is also the editor in chief of International Journal of Chronic Diseases & Therapy, an associate editor of EPMA Journal, Frontiers in Endocrinology, and BMC Medical Genomics, and a guest editor of Mass Spectrometry Reviews, Frontiers in Endocrinology, EPMA Journal, and Oxidative Medicine and Cellular Longevity. He has published more than 148 articles, 28 book chapters, 6 books, and 2 US patents in the field of clinical proteomics and biomarkers.",institutionString:"Shandong First Medical University",institution:{name:"Affiliated Hospital of Shandong Academy of Medical Sciences",country:{name:"China"}}}]}},subseries:{item:{id:"28",type:"subseries",title:"Animal Reproductive Biology and Technology",keywords:"Animal Reproduction, Artificial Insemination, Embryos, Cryopreservation, Conservation, Breeding, Epigenetics",scope:"The advances of knowledge on animal reproductive biology and technologies revolutionized livestock production. Artificial insemination, for example, was the first technology applied on a large scale, initially in dairy cattle and afterward applied to other species. Nowadays, embryo production and transfer are used commercially along with other technologies to modulate epigenetic regulation. Gene editing is also emerging as an innovative tool. This topic will discuss the potential use of these techniques, novel strategies, and lines of research in progress in the fields mentioned above.",coverUrl:"https://cdn.intechopen.com/series_topics/covers/28.jpg",hasOnlineFirst:!1,hasPublishedBooks:!0,annualVolume:11417,editor:{id:"177225",title:"Prof.",name:"Rosa Maria Lino Neto",middleName:null,surname:"Pereira",slug:"rosa-maria-lino-neto-pereira",fullName:"Rosa Maria Lino Neto Pereira",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bS9wkQAC/Profile_Picture_1624519982291",biography:"Rosa Maria Lino Neto Pereira (DVM, MsC, PhD and) is currently a researcher at the Genetic Resources and Biotechnology Unit of the National Institute of Agrarian and Veterinarian Research (INIAV, Portugal). She is the head of the Reproduction and Embryology Laboratories and was lecturer of Reproduction and Reproductive Biotechnologies at Veterinary Medicine Faculty. She has over 25 years of experience working in reproductive biology and biotechnology areas with a special emphasis on embryo and gamete cryopreservation, for research and animal genetic resources conservation, leading research projects with several peer-reviewed papers. Rosa Pereira is member of the ERFP-FAO Ex situ Working Group and of the Management Commission of the Portuguese Animal Germplasm Bank.",institutionString:"The National Institute for Agricultural and Veterinary Research. 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Behind these definitions are hidden all the aspects of normal and pathological functioning of all processes that the topic ‘Metabolism’ will cover within the Biochemistry Series. 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Thus proteomics, an area of research that detects all protein forms expressed in an organism, including splice isoforms and post-translational modifications, is more suitable than genomics for a comprehensive understanding of the biochemical processes that govern life. The most common proteomics applications are currently in the clinical field for the identification, in a variety of biological matrices, of biomarkers for diagnosis and therapeutic intervention of disorders. From the comparison of proteomic profiles of control and disease or different physiological states, which may emerge, changes in protein expression can provide new insights into the roles played by some proteins in human pathologies. Understanding how proteins function and interact with each other is another goal of proteomics that makes this approach even more intriguing. Specialized technology and expertise are required to assess the proteome of any biological sample. 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