The temperature and viscosity changes of DNA during the photodynamic process
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Photodynamic therapy (PDT) (also known as photoirradiation therapy, phototherapy or photochemotherapy) is a new modality for cancer treatment and involves the combination of light with a photosensitizing drug in an oxygen-rich environment [1, 2]. The photodynamic therapy has been successfully used in the last decade for the therapy of solid malignant tumors and non-tumoral diseases such as psoriasis, age-related macular degeneration, actinic keratosis, ageing [3]. The PDT mechanism could be a direct one (by apoptosis and necrosis), or an indirect one (by immune response on tumor vasculature). An ideal sensitizer should have a defined pure chemical structure, a proper absorption in red/near-infrared region, and a good capacity of singlet oxygen generation. After irradiation, the photosensitizer (PS) passes into the singlet oxygen excited state and afterwards it can pass into the triplet state; in this state it can react with molecular oxygen, generating singlet oxygen or it can react with different molecules from the tissue generating radical forms of PS [4]. This final form can react with oxygen, leading to the formation of different reactive oxygen species such as the hydroxyl radical, hydrogen peroxide and the superoxide anion, which in their turn may oxidize the macromolecular cellular components, resulting in cellular death through either apoptosis or necrosis. PDT induces oxidative stress at specific subcellular sites through the light activation of organelle-associated photosensitizers, and is used in the destruction of tumor cells.
This chapter offers the most up-to-date results of photodynamic therapy with synthetic sensitizers and/or coupled with some natural antioxidants, by using our data correlated with literature reports.
Photodynamic therapy (PDT) is a relatively new type of treatment for cancer, and makes use of a photosensitizer, visible light and molecular oxygen. Reactive oxygen species (ROS) are generated, causing the death of tumor cells by apoptosis or necrosis. Significant research efforts are nowadays focused on finding new photosensitizers with antineoplastic activity.
Over the last decade, there is an increasing interest in photosensitization mechanisms in biological systems, in relation to the therapeutic aspects of this phenomenon [5, 6]. The most used photosensitizers in photodynamic therapy are porphyrins, phthalocyanines and related compounds (Figure 1). These compounds are capable of generating a long-lived triplet excited state, responsible for facile energy transfer to molecular oxygen, leading to the formation of singlet oxygen. The photosensitization reactions include free radical reactions (type I) and singlet oxygen reactions (1O2) (type II) (Figure 2). The mechanism of PDT may lead to a direct tumor cell injury and also an indirect cell death via microcirculatory changes, resulting in reduced blood flow to the tumor [7-10].
The structure of porphyrin (left) and phthalocyanine (right). (R=meso-substituent; M=central metal)
The dual (up) and singlet mechanism (down) of PDT
The study of radical-induced damage in living systems is a topic of great interest in biology and medicine. Attention has been increasingly focused on the role of free radicals in normal physiological conditions and in different pathologies, with an essential role in cellular processes resulting in damages [11].
Porphyrins are used in medical and biological applications as they can generate new sensitizer structures with extensive and versatile photophysical and photochemical properties. Synthetic porphyrins and phthalocyanines appear to be good models in order to create new efficient drugs for photodynamic therapy of cancer [12]. The photodynamic therapy of cancer is based on the ability of some sensitizers to be retained in larger amounts and longer time in neoplastic than in normal cells as well as on the possibility to generate singlet oxygen after the activation of the porphyrin with red light [13].
In our previous investigations on PDT sensitizers [14, 15], we have used several spectroscopic methods to evaluate the efficiency of triplet state generation of some sensitizers [16]. The absorption spectra of the dye incorporated into the cells provide information with low accuracy because of small absorption amplitudes and perturbation of the spectra by light scattering. Therefore fluorescence methods are widely used [17]. The fluorescence of photosensitizers observed under a confocal microscope enables to establish the localization of dye in the cells [18–20]. Changes in the fluorescence of cell material due to illumination of samples depend on the sensitizer localization in the cells, but for a group of similar dye molecules, the cells, stained to a higher degree, usually show higher fluorescence intensity [21].
Within PDT, a sensitizer, light and oxygen are used to cause photochemically induced cell death via apoptotic pathways. In vitro and in vivo photosensitizing efficacy of certain synthesized photosensitizers are usually investigated by their interaction with some proteins (human serum albumin (HSA) (Site II) for binding affinity, intracellular localization and DNA attack [22, 23].
The photobiological mechanisms by which some sensitizers induce tumor necrosis are very different: they strongly associate with serum albumins and it is assumed that they kill neoplastic cells indirectly by damaging blood vessels and interrupting the supply of oxygen [24]. A good example is water-soluble porphyrins, tetrakis (4-sulfonatophenyl) porphyrin (H2TSPP) as one of the best tumor localizer. It has a very low affinity for human plasma lipoproteins but binds strongly to human serum albumin (HSA), Figure 3. The association between H2TSPP and HSA has a spectacular effect on the singlet-state lifetime of porphyrins: it decreases from 5.2 ns to about 1.6 ns [25]. This marked decrease is due to the enhancement of one of the molecular processes involved in the deactivation of the singlet state. The shortening of the singlet state lifetime causes an important decrease of the quantum yield of triplet state (ϕT). This assumption is in agreement with the result of Davila and Harriman [26] that measured the ϕT of the complex H2TSPP–HSA and reported a value of 0.70. The shortening of the singlet state lifetime of this complex is compensated by a corresponding increase of the intersystem crossing rate. The photosensitizing efficiency of bound H2TSPP has a reduced mobility, characteristic of molecules bound deeply into the protein structure. Such molecule is expected to be less accessible by oxygen and thus less efficient in producing singlet oxygen. The number of binding sites per protein molecule is greater than unity (
The changes of absorption spectra during the interaction between H2TSPP and HSA
On irradiation with light (laser), in the presence of a photosensitizer, DNA undergoes several modifications including chain breaks, DNA–protein cross-links, and basic sites changes coupled with oxidized DNA bases, which have been shown to be able to induce point mutations [27]. The single strand breaks (SSB) are mainly formed through the attack of OH radicals, where the hydrogen bonds between purine bases play an important role. OH radicals react with DNA to remove an H atom, leading to strand rupture. Mitochondria play a central role in the control of apoptosis induced by PDT, which through the mitochondrial permeability transition pore (PTP) lose the integrity of the outer mitochondrial membrane, thus releasing the intermembrane proteins, such as cytochrome
The porphyrins are able to induce changes of melting points and viscosity of DNA, which will be fragmented into small chains (Table 1). In this case, two modes of interaction between dye molecules and DNA are distinguished: a strong binding mode involving about 20-23% of the DNA phosphate groups and a weak binding mode involving the remaining phosphate groups.
\n\t\t\t\t | \n\t\t
The temperature and viscosity changes of DNA during the photodynamic process
An analysis on DNA degraded by sensitized attack of methylene blue (MB) showed that it had 80% of the guanine residues [29], and similar value for hematoporphyrins and furrocoumarins [30]. Apoptosis was evident in the post-PDT cells through the TdT-mediated dUTP nick end labeling (TUNEL) method and DNA fragmentation assay. Apoptosis was determined by cell morphology with light microscopy and transmission electron microscopy [31].
The two known modes of interaction between dye molecules and DNA are a strong binding mode involving about 20–23% of the DNA phosphate groups and a weak binding mode involving the remaining phosphate groups [32].
Based on the interaction sensitizer-DNA, three groups of porphyrins are known:
Group I porphyrins, which induce changes characteristic of intercalation in DNA samples with greater 40% GC base composition with an increasing of the linear viscosity of CT DNA, strongest intercalative binding in GC regions and the localization of AT regions only outside binding.
Group II porphyrins are believed to be outside binders, with an additional self-stacking features that induce DNA aggregation and a small viscosimetric changes of DNA with greater than 40% GC content.
Group III porphyrins give results characteristic of outside binding at both AT and GC sites, identified by the absence of an increasing of the solution viscosity for any of the linear or super helical DNA samples; they being able to bind with a preferential orientation.
Cationic porphyrins can interact with DNA either by intercalation between the G–C base-pairs or by outside binding in the minor groove with A–T sites selectively. An exemplification has been checked by personal data for cytosine (C) and guanine(G) by using as sensitizers methylene blue (MB), H2TSPP and Rhodamine (Rh) (Figure 4).
Cytosine (C)- and guanine(G)-sensitized photooxidation by using as sensitizers MB, H2TSPP and Rh
The weakly bound dye molecules are identified to be attached to the helix exterior by means of electrostatic interaction with the phosphate groups. A red-shifted absorption is due to porphyrin–porphyrin interactions within long-range assemblies on the polymer in exterior. When the porphyrin is intercalated, a strong hypochromicity and a strong bathochromic shift for the absorption bands are observed. As a possibility of external association a small hypochromicity (or even a hyperchromicity) and a small batochromic shift are observed. If an external association is possible in concert with an aggregation process, only hypochromicity (or hyperchromicity) is important [28]. Some important conclusions had been stated: porphyrins intercalate in DNA with a binding constant of around 103–106 M-1; the porphyrin complex appears to be stabilized by extensive electrostatic interactions, especially in the minor groove; the porphyrin does not form van der Waals stacking contacts with adjacent bases; DNA may require conformational distortion reaching the limit of DNA melting; the resulting severe conformational distortion not resolved upon achievement of the ground-state complex; steric clashing between the DNA backbone and the porphyrin extending the DNA along its helical axis; and de-stacking the interior of the complex.
The aggregation process is very important in this interaction study. If the sensitizer is in a monomer form, the external association with DNA could be favored. The association process increases its rigidity and changes the distance between the bases creating the adequate distances unfavorable to the dimerization. The Soret band is batochromically shifted from 412 nm to 433 nm, and the Q-band decreases its intensity. New bands appear at 644 and 707 nm. Also, a new band appears at 490 nm (Figure 5). The band at 490 nm arises from the J-aggregate (edge-to-edge interaction) of porphyrin molecules [33]. Their structures have been identified by atomic force microscopy (AFM) (Figure 6). The porphyrins are externally bound when their planar structure fit into the major or minor groove and interact with DNA electrostatically. Also, hyperchromicity is observed (the greatest absorption value at 260 nm), which is similar with some mono- and di-catenare destructions. It could be presumed that during the laser irradiation support a photooxidation reaction takes place preferably at guanine site. By irradiation with DNA, an increase of viscosity is observed as an effect of a chain-breaks and helix destruction.
The changes of absorption spectra during the interaction between H2TSPP and DNA
The structure of H2TSPP in monomer (left) and J-aggregate (right) form
The fluorescence emission spectra of H2TSPP, DNA and their complex consist of two bands with maxima little affected by DNA; however, the emission intensity is markedly reduced (Figure 7). For H2TSPP, in the presence of DNA, the maxima from 605 and 640 nm increase simultaneously, as a proof for the external binding between both compounds. From the amplitude ration I604/I640, it was concluded that the porphyrin/DNA this ratio increases without irradiation and by irradiation with laser beam, this ratio registered a significant increase at the high time of irradiation. A spatial arrangement of porphyrin and DNA takes place on the DNA strand exterior based on electrostatic interaction between phosphate groups of DNA and positive charges of J-aggregated porphyrin. If H2TSPP has a J-aggregate form, it is possible to cover DNA molecule like two concentric bodies. DNA has a 20 Å diameter, while H2TSPP -J-aggregate has 0.35–0.40 Å interplanar distances and a 40 Å diameter [34].
The fluorescence of H2TSPP and its mixture with DNA at different concentrations.
The most used and most efficient sensitizers are synthetic compounds, as porphyrins (P) and phthalocyanines (Pc).
Porphyrins constitute a class of the molecules that contain four pyrrole rings linked by the methane carbon bridges. A large group of porphyrins can be chemically modified by introducing the metal at the center of the pyrrole rings or by attaching the peripheral groups to the outer rings of the methane bridges units, respectively. The main characteristic of free-base porphyrin consists in the absorption maxima: a Soret band around 400 nm and four Q-bands in the region of 500–700 nm. Although the porphyrins absorb light poorly in this wavelength region (650 nm, ε= 30000 M-1⋅∙ cm-1) [35], as a result of increased transparency of biological tissues at longer wavelength; red light is normally used for PDT. Some exemplification of porphyrins and phthalocyanines are shown in Figure 8.
Phthalocyanines (Pc)/naphthalocyanines (Nc) are molecules composed of four indole units: pyrrole rings linked by nitrogen atoms conjugated with benzene rings characterized by a strong, isolated Q band in the red region of the UV/Vis spectrum, while the less intense B band is found at higher energies (Figure 9). The Q band is characterized by a high molar absorptivity (ε = 105 M-1 ⋅ cm-1) [36], and it is accompanied by a series of vibrational bands. The B band is broad due to the super-positioning of B1 and B2 [37].
The structure of some SiPc, SiNc and SiTPP
Porphyrins act as free bases and chelated with a variety of metals, the diamagnetic ones enhancing the phototoxicity. Paramagnetic metals shorten the lifetime of the triplet state and as result can make the dyes photoinactive [38]. The photosensitizing activity is quenched by the presence of transition metal ions (as central ions) with a d-electron configuration [39]. An exemplification is Si(enolate)2 5, 10, 15, 20-tetra-
The presence of axial ligands to the centrally coordinated metal ion is often advantageous, since it generates some degree of steric hindrance to intermolecular aggregation, without impairing the photophysical properties of the dye. Several photophysical parameters for tetrasulfonated aluminum porphyrins (Table 2) have been evaluated by means of some in vitro experiments on EL-4 cells [40].
The spectra of Pc as free base (a) and metallic complex (b)
\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t \n\t\t\t\t | \n\t\t\t\n\t\t\t\t \n\t\t\t\t | \n\t\t\t\n\t\t\t\t \n\t\t\t\t | \n\t\t
\n\t\t\t\t | \n\t\t\t215; 230; 263; 266; 432; 570; 603; 645; 827 | \n\t\t\t236; 254; 300; 330; 368; 438; 564; 644 | \n\t\t\t344; 425;588; 651; 676;708; 854; | \n\t\t\t950; 1075 | \n\t\t\t0.23 | \n\t\t
\n\t\t\t\t | \n\t\t\t212; 222; 264; 428; 564; 606; 645 | \n\t\t\t250; 300; 422; 564; 608 | \n\t\t\t426; 598; 652; 826; 706; 776; | \n\t\t\t950 | \n\t\t\t0.37 | \n\t\t
\n\t\t\t\t | \n\t\t\t212; 222; 264; 428; 564; 606; 645 | \n\t\t\t442; 644 | \n\t\t\t426; 598; 652; 856 | \n\t\t\t1075 | \n\t\t\t0.32 | \n\t\t
Spectral properties and data for partitioning coefficients of some metallo-porphyrins
The quantum yield for singlet oxygen generation was evaluated in this experiments by using DPBF method [41], and an exemplification is shown in Table 3.
\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t
C2AlTSPP | \n\t\t\t0, 96 | \n\t\t\t5.03 | \n\t\t
C4AlTSPP | \n\t\t\t0.78 | \n\t\t\t40, 8 | \n\t\t
C8AlTSPP | \n\t\t\t0.83 | \n\t\t\t32, 9 | \n\t\t
C12AlTSPP | \n\t\t\t0.665 | \n\t\t\t47, 5 | \n\t\t
The singlet oxygen quantum yields and the lifetime values for the first excited states of Al porphyrins
The cellular uptake of different drugs seems to be correlated to their hydrophobicity only when the drugs are very closely related to chemical structure. Aluminum ion has a great influence on the sensitizer hydrophobicity without a corresponding effect on the cellular uptake (Table 4).
\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t
TSPP | \n\t\t\t0.47 | \n\t\t
C2AlTSPP | \n\t\t\t0.23 | \n\t\t
C8AlTSPP | \n\t\t\t0.37 | \n\t\t
C12AlTSPP | \n\t\t\t0.32 | \n\t\t
The data for cellular uptake of the aluminum porphyrins
Nevertheless, C2AlTSPP is not as well uptaken as H2TSPP, even in the studied series, porphyrin is the most reactive.The uptake ratio of porphyrins in vitro increases when increasing the lipophilic property of the drug. The uptake of aluminum porphyrins increases linearly with increasing lipophilicity. The inactivation kinetics for EL-4 cells is shown in Figure 10 and their microscopic aspects are shown in Figure 11.
The inactivation diagram for EL-4 cells with aluminum porphyrins (1=AlC12TSPP; 2=AlC8TSPP; 3=AliBuTSPP; 4=AlC2TSPP; 5=Control
The microscopic aspects of cells before (left) and after PDT (middle and right)
Apoptosis (type I cell death) is different from necrosis (type III cell death) [42, 45]. If apoptosis is a controlled suicide pathway, involving DNA breaks, membrane blebbing, cell shrinkage and phagocytosis, necrosis involves membrane damage, local inflammation and injury, which lead to oncosis. Except of these types, there is type II cell death, which is characterized by an enormous increase of two-membrane autophagic vacuoles in the cytoplasm, which are finally catalyzed by lysosomal hydrolases [43]. Autophagy is a convertible process, which can provoke both survival and death pathways, in contrast to the apoptotic irreversible process leading only to cell death. Apoptotic cell death is the most preferable effect of various anticancer therapies, which leads to destruction and elimination of pathological cells. Inflammation does not occur through apoptosis in cancer cells and surroundings tissue [44].
PDT is a concerted action of a sensitizer, a light source in the presence of oxygen, based on singlet oxygen and ROS production, leading to cell death by different mechanism [46].
Reactive oxygen species (ROS) is a collective term used for a group of oxidants, which are either free radicals or molecular species able to generate free radicals. The most important free radicals occurring in the human body are hydroxyl radical (*OH), singlet oxygen (1O2) and superoxide (O2*-). Except these species, the intracellular generation of ROS mainly comprises nitric oxide (NO•) radicals, which together with O2•− radicals, are converted to powerful oxidizing radicals like hydroxyl radical (•OH), alkoxy radicals (RO•), peroxyl radicals (ROO•), singlet oxygen (1O2) by complex transformation reactions. Some of the radical species are converted into molecular oxidants like hydrogen peroxide (H2O2), peroxynitrite (ONOO–) and hypochlorous acid (HOCl). The oxidative stress damage is targeted mainly at cellular macromolecules, such as lipids, nucleic acids and proteins. Hydrophobic photosensitizers accumulate mainly in cell membranes and they are primarily attacked by free radicals. These oxidizing agents can damage cells by starting chain reactions such as lipid peroxidation, or oxidation of proteins or DNA. Damage to DNA can cause mutations and even major diseases (cancer), while protein oxidation can lead to distortion and degradation. Oxidation or oxidative stress leads to the production of free radicals, for example the hydrogen and oxygen molecules, which are not related to other factors. In their free form these molecules react with other molecules, and contribute to oxidative stress [47-49].
Oxidative stress, arising as a result of an imbalance between free radical production and antioxidant defenses, is associated with damage to a wide range of molecular species including lipids, proteins, and nucleic acids. Oxidative stress is a factor that initiates cell death after photodynamic reaction [50–54]. The formation of reactive oxygen species (ROS) with cytotoxic effects is the key involved in the death of the tumor cells by PDT [55]. PDT may be considered an oxidative stress that induces apoptosis in different types of cancerous cells both in vitro and in vivo [56].
Cells have a highly developed and regulated antioxidant defense system to maintain appropriate intracellular ROS levels and prevent oxidative damage. This system includes antioxidant enzymes such as superoxide dismutase (SOD), catalase and various peroxidases and non-enzymatic systems (GSH, thioredoxin, uric acid, vitamins, coenzyme Q) that effectively remove ROS. Under normal conditions, antioxidant mechanisms scavenge ROS and protect the organism from the damaging effects of ROS. However, under conditions of excessive oxidative stress, cellular antioxidant mechanisms may be unable to prevent the adverse impact of ROS on critical cellular processes. ROS can interact with cellular macromolecules, including DNA, protein and lipids, and interfere with vital cellular functions [35]. Mutations caused by ROS can result in malignant transformation and the development of cancer [57].
To evaluate the presence of oxidative stress in the plasma, we used indirect methods. These quantify the lesions produced by the reactive oxygen species on the organism’s biomolecules.
To evaluate the presence of oxidative stress in PDT, some of the following methods could be used [58–64]:
Detection of malondialdehyde (MDA), the marker used most frequently for lipid peroxidation, using the fluorimetric method with 2-thiobarbituric acid (TBA);
The carbonylated proteins (CP) with 2, 4-dinitrophenylhydrazine using the Reznick method. To determine the level of proteins from the homogenate samples Bradford method was used.
The hydrogen-donating capacity (HDA) using the stable free radical: 1, 1 diphenyl-picrylhydrasyl (DPPH);
Detection of the –SH groups with 2, 2-dithiobisnitrobenzoic acid (Ellman’s reagent). The zymographic method was used to determine the MMPs activity.
Some synthetic PSs such as porphyrins have the capacity to generate reactive oxygen species with cytotoxic effects leading to the necrosis of tumor cells, and induced a significant oxidative stress response, with peak intensity at 24-hour post-exposure. The most significant responses were the increase of CP, MDA, MMP-2 (matrix metalloproteinase-2) activity and the decrease in HDA levels. The experiment also induced less significant, but present, reduction of the thiol groups.
As an exemplification, PDT with H2TSPP increases ROS production in plasma and tumor tissue and determines oxidative alterations of biomolecules (lipids, proteins) (Figure 12).
Oxidative stress, responsible for tissues injured in different pathologies, involves the nonequilibrium between the produced radical species and antioxidant defense agents.
There are some priority parameters which should be determined and evaluated:
Cell viability test measured by the lactate dehydrogenase release in the supernatant of cell culture as a marker of cell integrity. Viability results were expressed as % of live cells from the suspension subjected to 24 h incubation in the tested agents and/or irradiation.
Cell proliferation test measuring the number of viable cells as live cells reduce the kit reagent into a formazan compound that is colorimetric measured at 490 nm. From this point of view the test measures the quantity of viable cells in culture and thus the proliferative capacity of the tested cells.
RNA synthesis achieved by tritium-labeled uridine incorporation method, which involves beta-radioactivity measurement for radiolabeled cell cultures.
Total cellular RNA offer the concentration of total RNA, measured with SV total RNA isolation system.
TUNEL and immunofluorescence essential for apoptosis detection by observing the DNA fragmentation assay and TdT-mediated dUTP nick-end labeling (TUNEL) assay. TUNEL staining was performed to detect internal and end-strand breaks, which often occur in the early stages of apoptosis. TUNEL staining was carried out according to the manufacturer’s instructions (Promega, Madison, USA). The procedure is carried out as follows. Biotinylated nucleotide is incorporated at the 3\'-OH DNA ends using the terminal deoxynucleotidyl transferase (rTdT) recombinant enzyme. The apoptotic cells were counted under the microscope and photographed.
Analysis of genomic DNA fragmentation visualized by staining with ethidium bromide (0.5 µg/ml) and photographed under UV illumination. DNA was separated using standard 1.5% agarose (Bio-Rad) gel electrophoresis at 10 V/cm.
Cellular morphology visible by electron microscopy necessary for identifying the apoptotic cells. After PDT treatment, cells are developing morphological features characteristic of apoptosis: chromatin compaction into uniform electron-dense masses with nuclear margination, nuclear fragmentation, cellular shrinkage, cell membrane vacuolization and blebbing, and the increase in electron-density of the cytosol [63-65].
Photodegradation reaction of sensitizers, which are indicators for lifetime of the sensitizers, time efficiency and potential side effects of them (Figure 13).
The oxidative stress parameters for PDT with H2TSPP. Adapted after [
Some of these parameters have been calculated for PDT with different sensitizers and for different cells; B16, K562, EL-4, etc. [64–69]. During irradiation, after the first 30 minutes, a strong cellular degradation, especially for TNP-loaded cells, is visible (Figure 14). The results indicate that a free-base porphyrin as 5, 10, 15, 20-tetra-
The photo degradation scheme of a sensitizer
Time evolution of the cellular density for TNP-loaded K562 cells (10 µg/ml TNP)
After irradiation, the K562 cell viability is strongly affected both for TNP-loaded cells and for those unloaded. TNP activated increases the cell mortality by comparison with control cells (Mi) (Figure 15).
The viability of K562 loaded with TNP after the first hours after irradiation
At different time post-irradiation, TNP intracellular loaded and activated shows a homogeneous effect on the cells’ capacity to deliver LDH (Figure 16).
Time evolution of K562 cells viability after laser activation in the presence of TNP
RNA isolation from K562 cells has been achieved in the first 4 hours after irradiation. A 35% decrease of RNA content has been registered in the case of H2TSPP (Figure 17) [72], Also, there is a good correlation between uridine incorporation and number of live cells in 24 h post-irradiation in the presence of the same sensitizer, both of them decreading (Figure 18). Also, the amount of total cellular RNA isolated from the K562 cells drastically decreased after irradiation (Figure 19).
The viability and multiplication rate of K562 tumor cells at 24h post-irradiation. C = nonirradiated unloaded cells; TSPP = nonirradiated loaded cells; Ci = irradiated unloaded cells; TSPPi = irradiated loaded cells.
At different time intervals after treatment, the cells were analyzed for caspase-3. At 4 h post-PDT, 100% of the cells displayed protease activities. After PDT, 30.6% of the cells showed total caspase activity. Thereafter, the fraction of cells with caspase activity increased to 48.6%.
When present in the cytoplasm, a number of caspases have been activated following PDT and responsible for the cleavage of multiple cellular proteins, DNA fragmentation, and cell death. Activation of procaspase-3 after PDT has been demonstrated in multiple experimental settings. The morphological manifestation of apoptosis (“execution” phase) can be ascribed as degradation of various structural proteins and DNA. After loading K562 cells with TNP, and irradiation, a strong decrease has been obtained for caspase-3 activity (Figure 20).
Correlation between uridine incorporation and number of live cells in 24 h post-irradiation. C = non-irradiated unloaded cells; Ci = irradiated unloaded cells; H2TSPPi = irradiated loaded cells.
The amount of total cellular RNA isolated from the K562 cells. C = unloaded non-irradiated cells; Ci = irradiated unloaded cells; TSPP = non-irradiated loaded cells; TSPPi = irradiated loaded cells.
Caspase 3 activity in K562 cells loaded with 10 μg/ml TNP and activation by irradiation.
Induced apoptosis of K562 cells loaded with 10 μg/ml TNP and activated with laser (TUNEL) has been shown in Figure 21, as results of TUNEL test. Many apoptotic cells characterized with brown nuclei can be seen in the TNP-PDT groups.
The PDT efficacy relies on the concerted action of sensitizer and light, with none of them alone inducing apoptosis.
Induced apoptosis of K562 cells loaded with 10 μg/ml TNP and activated with laser (TUNEL). Results of TUNEL. Many apoptotic cells characterized with brown nuclei can be seen in the TNP-PDT groups.
An antioxidant is a molecule stable able to donate an electron to a free radical, neutralizing it, thus reducing its capacity to damage. The antioxidants delay or inhibit cellular damage mainly through their free radical scavenging property. A variety of dietary plants including grams, legumes, fruits, vegetables, tea, wine etc. contain antioxidants. The most important antioxidants seem to be nonenzymatic antioxidants derived from plant sources including vitamins (vitamin A, C, E, K), flavonoids (quercetin, catechin, epigallocatechin gallate, hesperidin, hesperetin, diosmin, and many others), phenolic acids (cinnamic acid derivatives, curcumin, caffeine, catechins, gallic acid derivatives, salicylic acid derivatives, chlorogenic acid, resveratrol, folate, anthocyanins and tannins) [74–80].
Carotenoids, known as naturally fat-soluble pigments and synthesized by vegetal organisms, are sources of different colors [81]. They could be classified into carotenes (beta-carotene and lycopene) and xanthophylls (lutein and zeaxanthin).Their structure is shown in Figure 22. Beta-carotene and lycopene are widely regarded as being effective antioxidants, with small sizes (nm), specific absorption and fluorescence spectra, and are easy to detect (Figures 23–25). The synergic antioxidant effect of the mixture lycopene-beta-carotene-vitamin E on some cellular systems (in vitro and in vivo), has been reported [82–84]. Some antioxidants with health-protective effects (lycopene, beta-carotene, vitamin C, quercetin-glycosides, naringenin-chalcone, chlorogenic acid) are seen in tomato plants.
From ancient times, plants have been used intuitively for medicinal purposes. A large number of plants have been investigated and various species have been reported to exhibit antioxidant activity, including Marigold flower (
The transformation diagram of lycopene
The DLS measurement of lycopene
The absorption (left) and fluorescence (right) spectra of lycopene
The absorption spectra of lycopene (- - -) and carotene (_____)
Many antioxidants could be identified in tea leaves and fruits. For example, green tea, produced from the leaves of the plant (
The chemical structure of the main epicatechin derivatives/polyphenols from green tea
The antioxidant activity (AA%) of the studied samples and their inhibitory effect against free radicals was evaluated using the DPPH method, by used the following formula:
where: Acontrol is the absorbance of a DPPH solution without sample, Asample is the absorbance of the sample mixed with DPPH solution.
Some citrus extracts are very important for their antioxidant activity such as hesperidin, hesperitin and diosmin (Figure 27). Among the flavonoids used in oral administration in chronic leg ulcer, hesperidin is a glucozid that is abundant in citrus fruits. Recently, formulating hesperidin in nanocrystals, has provided its dermatological application, assessing its antioxidant effect. In vitro studies have shown its clear antioxidant properties, and using them as nutrients has shown its vaso-protective action.
The structure of hesperidin, diosmin and hesperetin
Among mostly exploited flavonoids in chronic venous condition, hesperidin (C28H34O15) [+/−-3, 5, 7-trihydroxy-4ʹ-methoxyflavanone 7-rhamnoglucoside] is a flavanone glycoside group of flavonoids found in large amounts in citrus fruits, grapefruit peels, lemon, oranges, blond grapefruit (
The absorption spectra of hesperidin, hesperetin and diosmin
For all of them, total flavonoids content (TFC), total poliphenols content (TPC) and antioxidant activity (AA
\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t
Diosmin etalon in MeOH | \n\t\t\t85.53 | \n\t\t\t521.186 | \n\t\t\t55.79 | \n\t\t
Hesperidin etalon in MeOH | \n\t\t\t43.1 | \n\t\t\t466 | \n\t\t\t59.31 | \n\t\t
Hesperetin etalon in MeOH | \n\t\t\t169.16 | \n\t\t\t1814.406 | \n\t\t\t73.34 | \n\t\t
TFC, TPC and AA content
All the above measured parameters show very close values to the literature reports [106].
In spite of numerous advantages, the photodynamic therapy has a number of limitations. The light source should be close to the appropriate site in order to be effective before the diagnostic to be established. Persistent skin photosensitivity is observed some weeks after the treatment, which is considered as the main side effect. Also, this method is not possible without a light source and therefore entails high costs for the whole treatment. PDT acts both on ill cells and in a small manner on healthy cells. The intracellular Ca2+ from the cells induce low levels of shear stress on them, without any morphological changes [112]. The reactive oxygen species are localized to the cancer cells selectively destroying them and not the surrounding normal tissue. The presence of the lymphoid cells in tumor immunity has been demonstrated during PDT with various photosensitizers. The immunologic effects include the production of interleukin 1-beta, interleukin 2, tumor necrosis factor-alpha, and granulocyte colony-stimulating factor. PDT acts to induce oxidative stress by the generation of free radicals to damage DNA and proteins, and eventually cell death, by necrosis and apoptosis. The reactive oxygen species lead to the state called oxidative stress. The antioxidants offer protection against lipid oxidation, react and interfere with free radicals, reduce oxidative stress, and stop low-density lipoproteins from being oxidized. Also, they protect healthy tissues and lower the incidence of treatment-related side effects. In the context where many clinical studies with respect to the application of antioxidants as sensitizers are lacking, this chapter showed a systematic review by showing evidence of the antioxidant action in photochemotherapy and their comparison with synthetic sensitizers (porphyrins and phthalocyanines). Under such circumstances, PDT is extremely important for the treatment of different diseases: lung, bladder, and skin cancers (precancerous and even melanoma).
We have received the financial support of the project PNII 185/2014 to submit this book chapter.
Observations of crystal-like ordering of fine particles in plasmas were first reported in 1994 [1, 2, 3, 4]. They were performed by growing carbon particles in a methane plasma [1], silica particles growing in plasma [2], and ready-made polymer particles in argon plasmas [3, 4]. The possibility of observation of crystal-like ordering of fine particles, which was called Coulomb solid or Coulomb crystal, in a low-pressure plasma was predicted through calculation by Ikezi [5]. It was similar to those in the solution of colloids generally negatively single-charged, while micron-sized fine particles get thousands of electrons or more on their surface in a plasma. They can form solid state easily because of larger value of the Coulomb-coupling parameter, Γ, as follows:
where
Charge neutrality is satisfied in a fine particle plasma as
where
The physics of one-component plasma or strongly coupled plasma has been studied mainly by computer simulation such as the Monte Carlo method and molecular dynamics since 1957, when the liquid–solid phase transition by hard spheres in a confined condition was discovered by Alder and others [7, 8, 9]. Following this study, the phase transitions were confirmed using other repulsive potentials such as Coulomb [10], soft sphere [11], and Yukawa [12, 13].
Although theoretical or computer-simulation studies on one-component plasma and strongly coupled plasma have been proceeded, there are not many experimental studies except for the use of ion trap or colloidal solutions. Ions confined in a Pawl or Penning trap can form crystal ordering [14, 15]. Colloids in electrolyte solution form crystal structures [16]. However, special skills and time are required for the formation. Compared to that, Coulomb crystals in fine particle plasma are formed more easily and quickly.
In this chapter, the formation, observation, and analysis of Coulomb crystals in fine particle plasmas are presented. Current status of their application to study of solid state physics phenomena is also described.
Coulomb crystals in fine particle plasmas have been formed typically under the conditions as shown in Table 1.
Fine particles | Material | Polymer, silica, carbon |
Shape | Sphere | |
Size | 1–10 μm | |
Size distribution | Monodisperse | |
Discharge device | Power supply | RF (13.56 MHz), DC |
Electrode type | Parallel-plate | |
Plasma conditions | Discharge gas | Argon, helium, methane |
Pressure | 10–100 Pa | |
Discharge power | As low as limit of particle suspension |
Typical conditions for Coulomb crystal formation in plasma.
Figure 1 shows an example of a plasma system for Coulomb crystal formation [17]. The system is a parallel-plate 13.56-MHz radio-frequency (RF) plasma device, which includes eight pieces of permanent magnets in the RF electrode to generate planar-magnetron plasma. Discharge gas is introduced and evacuated far from the area of center of vacuum chamber in order that fine particles are suspended under a stagnant condition to reduce the effect of gas drag. A side view of suspended fine particles above the RF electrode is shown in Figure 2. In this case, fine particles of spherical divinylbenzene polymer 2.74 μm in diameter were injected into argon plasma under the pressure of 65 Pa and RF power of 2 W.
Schematic view of plasma system for Coulomb crystal formation. Dark red and light blue parts show RF electrode and viewing glass windows, respectively. Other parts are grounded.
Side view of arrangement of fine particles above RF electrode.
The arrangement of fine particles in Figure 2 is divided up and down. The upper part of fine particles forms horizontal planes, while the lower part forms vertical strings. It is judged that the border of the two particle arrangements corresponds to the plasma-sheath boundary. It was explained by theoretical analysis [18] and computer simulation [19] that the vertical particle string of fine particles in the lower part is created by ion-wake field, which is effective in the region of ion speed more than Mach sound velocity, that is, in the sheath region. When the particle strings are closely packed, they form a two-dimensional (2D) hexagonal structure. Meanwhile, the horizontal layers of fine particles in the upper part are formed under the one-dimensional (1D) compressive force in the vertical direction [20]. Fine particles are arranged closely packing in each plane. To minimize the free energy in three-dimension (3D), such particles tend to be staggered with those in the next layer forming 3D close-packed structures: hexagonal close-packing (hcp) or face-centered cube (fcc). In this way, fine particles can form a 2D hexagonal structure and 3D structures such as fcc and hcp. The possibility of formation of body-centered cubic (bcc) structure will be described in Section 2.2.
Coulomb crystals can be observed by laser light scattering. When a visible light laser is used, light scattering from micron-sized fine particle is in the Mie-scattering regime. In such a regime, fine particles can be individually observed by the scattered laser light of output power of even a few mW when they are in the arrangement of solid-like state.
3D structures of Coulomb crystal were observed by the use of plasma system as shown in Figure 3. The system consists of a vacuum reaction chamber, a blue argon-ion laser (wavelength: 488 nm, 100 mW output power), and two CCD video cameras, and a rotating analyzer for Mie-scattering ellipsometry [21]. In case of confirmation of the position of fine particles in the vertical direction, a red diode laser (wavelength: 690 nm, 20 mW output power) was also used. The light beam of the diode laser was expanded perpendicular to the RF electrode with the use of a cylindrical lens.
Schematic of plasma system for observation of 3D structure of Coulomb crystal [
The vacuum chamber is a cube with a side length of 10 cm. Viewing windows are provided in the five faces of the chamber. Two are for the incidence and outgoing of laser light. Two others are for the observation of Coulomb crystals from the side and the top. The other one is for the monitoring of the particle diameter by Mie-scattering ellipsometry. RF of 13.56 MHz was applied to an electrode of 4 cm diameter with the chamber wall grounded. A ring of 3 cm inner diameter was put on the electrode in order to effectively trap fine particles above it by forming a potential bucket. Gas inlet and exhausting ports were provided close to each other so that particles were not transported by gas flow.
Fine particles were prepared by film growth on the seeds of ultrafine carbon particles, which were injected at the first stage of the growth, in a 20% methane/argon plasma under the conditions of 5 W RF power and 40 Pa (0.3 Torr) pressure. Spherical and monodisperse carbon particles can be grown through coating of hydrogenated amorphous carbon on the seeds by the dissociation of methane gas in the plasma [22]. Using Mie-scattering ellipsometry, the diameter of growing particles was monitored, and the growth was stopped by the decrease of RF power to 1 W for the particles of diameter of 1.4 μm, with which three-dimensional Coulomb crystals were formed in the experiments that will be shown in Section 3.2.
Fine particles spread over the entire region in the potential bucket are suspended about 5 mm above the electrode in the luminous plasma region of the negative glow. The top and side images, which were taken at the same position and at the same time by irradiation with blue laser light only, of a 3D Coulomb crystal formed by 1.4-μm carbon particles are shown in Figure 4 [23]. The relative horizontal positions of fine particles in the top view were adjusted so as to agree with those in the side view at a crystal-grain boundary. Bright spots in the top view image indicate particles in the lowest layer and comparatively dimmed or small spots indicate those in the second lowest layer, because particles were illuminated in the off axis of the laser beam of the Gaussian distribution under them. From the correspondence of particle arrangement in the top view with that in the side view, particles are found to be aligned in the perpendicular direction of the side view plane. This means that each bright spot in the side view shows particles piled up in the direction (see the lines on the right in the figure).
Top and side video images of a 3D Coulomb crystal taken at the same time and the same position [
It can be seen that the crystal structure in Figure 4 is similar to the body-centered cubic (bcc) structure with (110) planes parallel to the electrode. The three-dimensional structure of a unit cell of the crystal is depicted in Figure 5. The lattice constants
3D crystal structures of bcc, bct, or fco (a) [
3D observation of the change of structure of Coulomb crystal was carried out. Figure 6 shows the top and side views of change of fine particles arrangement every 1/3 s. As the blue argon-ion laser beam runs through the lower region of fine particles and the scattered blue light is more intensive than the scattered red diode-laser light, the particle arrangement in a few lowest layers was distinguished in top views. Side views were taken by the use of another CCD video camera with an optical band-pass filter for the red light (690 nm) put in front of it. Therefore particles in a few vertical layers were observed by the latter CCD camera.
Structural transition of Coulomb crystal every 1/3 s from A to D. Side and top views were taken at the same time. Fine particles in the red colored regions in the top view correspond to those in the side view [
The crystal structure of stage “A” in Figure 6 was analyzed to be fco or bct, while that of stage “D” was fcc [21]. Change from “A” to “D” in the side view indicates that from the crystal observation axis of bct[100] to fcc[110] for, and change from “A” to “D” in the top view shows that from bct[110] to fcc[111]. The change of inclination of particle rows in the vertical direction is clearly seen in the side view. Corresponding to the side view, the particle arrangement in horizontal layers shown in the top view changes in the way that particles in the third lowest layer, which are indicated by very dimmed spots, gradually separate from the particle positions in the lowest layer. The change of crystal structure is illustrated in Figure 7.
Illustration of structural change of Coulomb crystal between bct (fco) and fcc from top view. Fine particles in the first, second, and third lowest layers are indicated by dark (blue), pale (green), and light (yellow) circles, respectively.
The results of the observation of the time and space changes of 3D Coulomb crystal structures suggest that the structural transitions occurred by the slip of crystal planes parallel to the electrode. The slip planes were fcc(111) and bct(110). In the martensitic transformation of metallic iron, the slip of crystal planes generally occurs between fcc(111) and bcc(110) or bct(110) [24]. The structural transitions of 3D Coulomb crystals in dusty plasmas well matches those of real metallic crystals.
When a crystal structure simply changes by slip of crystal planes from fcc to another cubic structure, the relation between two horizontal lattice constants should hold as
The intensity of light scattered by a single spherical particle is calculated through the Mie-scattering theory [25, 26] when the diameter,
The complex scattering amplitude functions,
where
In an analogous way to the reflection ellipsometry, angle parameters for Mie scattering are defined from the ratio between two complex scattering amplitude in the direction parallel and perpendicular to the scattering plane. When particles are spherical and monodisperse, the ellipsometric parameters Ψ and Δ are defined by the ratio of the scattering amplitude functions of
It is obvious that tanΨ = |
Figure 8 shows a (Ψ, Δ) trajectory as a result of Mie-scattering ellipsometry measurement for spherical carbon particles growing by coating in a plasma including methane (Figure 8(A)). The time evolution of diameter and density of fine particles are obtained from correspondence of graphs of (a) and (b) in Figure 8(B) as shown in Figure 9 [1].
Experimental result of evolution of ellipsometric parameters, (Ψ, Δ), during carbon particle growth. (A); (a) time evolution of Δ by experiment, (b) size evolution of Δ by simulated calculation (B) [
Time evolution of particle diameter (open circles and broken curves) and density (closed circles and solid lines) [
The transitions of arrangement of fine particles during the growth in a methane/helium plasma under the conditions of pressure of 106 Pa (0.8 Torr) and RF power of 5 W are shown in Figure 10 [34]. By Mie-scattering ellipsometry, particle diameter was evaluated as 1.3, 1.7, 2.2 μm at 30, 45, and 80 min, respectively. The particle arrangement at 30 min shows 3D regularity in the lowest four or five horizontal layers, especially crystal-like ordering in the lowest two layers. It is seen at 45 min that particles tend to be weakly chained in the vertical direction in the upper region. In its video movie, the chains were slowly swinging from right to left. The top view shows that particles in the lowest layer are randomly arranged. At 80 min, the particles are aligned like stiff rods in the vertical direction. They seem to form a close-packed 2D crystal, that is, hexagonal crystal. The structures of arrangement of smaller and larger particles correspond to those of upper and lower parts in Figure 2, respectively. However, the crystal edge of the 2D crystal is seen at the lowest side in the side view in Figure 10, while it is seen at the top side of 2D structure, which is the boundary between 2D and 3D structures, in Figure 2.
Changes of top and side views of particle arrangement (a). The particles of top views are in the two lowest layers and those of side views in several lowest layers. Illustrations of change of side-view particle arrangement are shown (b) [
The spatial distribution of size of arranged particles under the same conditions as shown in Figure 2 was analyzed by Mie-scattering ellipsometry using an image sensor instead of a photo-detector [35, 36]. Determined ellipsometric parameters are plotted on the Ψ-Δ coordinate plane, as shown in Figure 11, along with the trajectory obtained by calculation for spherical particles of refractive index of 1.56, the scattering angle of 88°, and diameter of 2650–2830 nm for each 10 nm. It should be noted that the trajectory shows a significant change with the diameter. The calculated trajectory was obtained for the best fitting to the determined values. By the comparison of the determined values Ψ and Δ with calculation, the size was evaluated to be 2.70, 2.74, 2.75, and 2.77 μm for particles in upper to lower layers (L4 to L1). Thus larger particles sank in lower position. However, although interlayer distance was observed to be about 100 μm, the distance between force balance positions for isolated 2.74 and 2.75 μm fine particles calculated in a way as shown in Section 4.2 is one order of magnitude smaller than the observed interlayer distance. The interlayer distance of 100 μm must be due to mutual Coulomb repulsion. The reason why larger particles exist in lower layers is explained as follows.
Ellipsometric parameters of measurement for fine particles in layers upper to lower: L1, L2, L3, L4 (closed circles and dotted line) and calculation for spherical particles of refractive index of 1.56 and diameter of 2650–2830 nm for each 10 nm (open circles and solid line).
The total chemical potential
where
where
Fine particles are suspended mainly at the balanced position of electrostatic force (
where
The three forces were calculated under the following simple assumptions:
Variation of electrostatic force (fE), ion-drag force (fI), and gravity (fG) with particle diameter.
Ion drag force could be larger than that calculated by Eq. (10) [37] because of more widespread ion-particle interaction beyond the Debye-length, which will be described in Section 4.2.
As was described in Section 4.1, the electrostatic, ion drag, and gravitational forces act on fine particles. The gravitational force pulls fine particles downward. When fine particles are suspended in a plasma generated between two parallel plates placed horizontally, they sink around the lower plasma-sheath boundary. If gravitational force is extremely decreased or lost, the position of force balance changes. Thus, microgravity experiments should be useful for analyzing forces acting on fine particles in a plasma.
A parallel-plate RF plasma system was used for the experiments (Figure 13) [38]. A piezoelectric vibrator for the injection of fine particles into a plasma was contained in an RF electrode, which was put at the bottom of vacuum chamber of the system. The top of the electrode is covered with a grid so that fine particles can be pushed up. A grounded counter electrode is placed at the upper side of the RF electrode at the distance of 14 mm with a vertically symmetric structure. The outer diameter of the two electrodes is 40 mm and the gap space of the electrodes is surrounded by a transparent plastic cylinder with an inner diameter of 35 mm. The vacuum chamber is shaped octagonal and has six viewing windows: one for the entrance of the laser light, another for the exit, and the other four for observation of the arrangement of fine particles using scattered laser light. Using two charge-coupled device (CCD) video cameras placed in front of two of the four windows, the XYZ-3D position of each fine particle can be determined.
Schematic of parallel-plate RF plasma system for microgravity experiment [
Spherical divinylbenzene fine particles 2.27 ± 0.10 μm in diameter were put into the piezoelectric vibrator. The density of the fine particle is 1.19 g/cm3, and the weight is 7.29 × 10−12 gw. Helium gas was introduced at the pressure of 133 Pa (1 Torr). After plasma was generated with the RF power of 2 W, fine particles were pushed up into the plasma. When a sufficient quantity of fine particles was introduced into the plasma, the vibrator was switched off and then the experimental system was placed in a microgravity environment.
Figure 14 shows images taken using the Y-axis video camera before and during the microgravity condition of less than 10−4 G (gravity constant) that was generated in a capsule of drop tower for 4.5 s at the Micro-Gravity Laboratory (MGLAB) at Toki in Japan. The number of injected fine particles was approximately 30,000. A diode laser, which radiates light 690 nm in wavelength, was used for the observation of fine particles by scattered laser light. The image obtained immediately before the drop under gravity shows fine particles aligning in a nearly vertical direction. The image obtained 2/30 s after the beginning of the drop shows the fine particles going straight up in the vertical direction, while the 6/30 s image shows the particles radially moving upward. In 1.5-s movie, the movement slowed down and the alignment of the fine particles was observed again, however, it was not vertical but radial. Fine particles aligned in radial directions are seen in the image shown at 3 s in Figure 14.
Evolutions of ordering of fine particles during drop experiment: (a) just before drop under 1 g; (b) 2/30 s after beginning of drop; (c) 6/30 s after; (d) 3 s after. Directions of fine-particles alignment and tilt angles are indicated in (a) and (d). The height and width of the figures are 14 mm and 21 mm, respectively [
The alignment of fine particles in the sheath should be caused by the effect of the formation of the wake field [18, 19]. The reason why the fine particles moved up after the beginning of the drop is due to the change in the balance of forces. Fine particles are suspended around the balance position of the resultant force (F for upward direction), which is composed of ion drag (
Illustration of upward resultant force composed of ion drag, electrostatic and gravitational forces and behavior of fine particle in drop experiment under 1 G (black broken curve) to 0 G (red solid curve) (a); potential energy calculated from integral of curves of the resultant force in (a) (b). Closed circles show force balance position under 1 G and upper position under 0 G; open circle shows upward force under 0 G at the position of force balance under 1 G.
Using the images shown in Figure 14 along with images taken at the same time with the X-axis video camera, three-dimensional particle position coordinates were obtained. Three-dimensional tilt angles were determined for six inner particles to be 6.4°and 3.2°for the left and right rows in Figure 14(a), and 30.2° and 30.1° for those in Figure 14(d), respectively. Interparticle distances between the inner first and second particles were 320 and 230 μm for the left and right rows in Figure 14(a), and 280 and 180 μm for those in Figure 14(d), respectively, and the Mach numbers of ion speed were calculated to be in the range from 1.2 to 1.6.
Since the structure of the RF plasma system is symmetric both vertically and horizontally, the shape of plasma generated in the plastic cylinder is supposed to be symmetric, that is, spheroidal, when the sheath thickness is comparable to the order of plasma size. Therefore, under zero gravity, fine particles are subjected to a resultant force in the spheroidal plasma, that is, they are affected by the positive ion flow and electric field whose directions are radial. If the wake field theory is accepted here, the radial alignment of fine particles can be reasonably understood as being due to the formation of rows in the direction of ion flow. On the other hand, the vertical alignment of fine particles under gravity cannot be explained with the assumption of a spheroidal plasma. In order to determine the shape of the plasma, we analyzed the density distribution of the plasma containing fine particles and drew 16 contours through the division of optical emission brightness on CCD images. As is shown in Figure 16, the plasma was lowered and flattened under gravity compared to that under microgravity. The results suggest that fine particles suspended around the lower plasma-sheath boundary deformed the plasma.
Contours of intensity distribution of optical emission from plasma containing fine particles under 1 G (a) and microgravity (b). The height and width of the figures are 14 mm and 19 mm, respectively. [
By the effect of plasma wake [18, 19, 42], a negatively charged fine particle forms a positive potential in the downstream position of ion flow, and another fine particle is attracted to the position of maximum potential. At the same time, the gravitational force pulls the downstream fine particle down from its position [43]. The third downstream fine particle that is downstream to the second one is also pulled down by gravity to a position lower than that of maximum potential. Such force actions are repeated on other particles further downstream. Due to the sinking of the fine particles under gravity, ion flow is affected by the negative potential formed by fine particles in turn and tends to be vertically directed lowering the free energy. As a result, under gravity, the mutual influence between the positive ion flow and the negative fine particles results in the alignment being more vertical and in the deformation of the plasma. The reason why fine particles moved straight up in the vertical direction, as shown in the 2/30 s image of Figure 14(b), is considered to be that the resultant force due to ion flow and the electric field was directed vertically upward just immediately after the diminishment of gravity.
Using the same plasma system that was used for drop experiment, a microgravity experiment by parabolic flight was carried out under the condition of less than 0.04 G in the vertical direction and less than 0.01 G in the horizontal direction for approximately 20 s at the Diamond Air Service Inc. in Japan. The plasma system was installed in a standardized rack in a jet plane. Figure 17 shows the images of fine particle behavior taken by the Y-axis video camera during the changes in gravitational conditions from 2 G (a) to microgravity (c) and to 1.5 G (e). Under microgravity, in Figure 17(c), fine particles are also observed around the lower balance position forming a void in the center as was reported [44]. The number of injected fine particles was approximately 10 times greater than that for the drop experiment. Most of the fine particles were pushed to the upper balance region at the moment of gravitational change, however, some remained behind due to their mutual Coulomb repulsive forces. In addition, fine particles injected continuously during the microgravity condition were also suspended around the lower balance position. Fine particles at the inner boundary were arranged parallel to the electrodes under gravity more than 1 G, while they formed an arc under microgravity even around the lower plasma-sheath boundary. In transition from 2 G to microgravity (b), the fine particles moved vertically upward. On the other hand, in transition from microgravity to 1.5 G (d), they moved downward through the outer side of the plasma and a void was formed in the center. The nonsymmetrical behavior of fine particles in transition can be explained by the difference in the state of plasma containing fine particles. As explained above concerning the results of the drop experiment, the resultant of ion drag and electrostatic forces pushed the fine particles vertically upward at the moment shown in Figure 17(b). However, the particles were obliquely pulled down during the increase of gravity as shown in Figure 17(d) by the oblique force that is composed of the gravitational force and the outward radial force, which is derived from the ion drag force stronger than the electrostatic force.
Video images of fine particle behavior in parabolic flight under gravitational conditions: (a) 2 G, (b) transition from 2 G to microgravity, (c) microgravity, (d) transition from microgravity to 1.5 G and (e) 1.5 G. In the case of (a), fine particles were accelerated by 0.1–0.2 G in the right direction of the image [
Such “void” was observed in the center of plasma under microgravity condition performed on the International Space Station (ISS) [45]. The cause of “void” formation is generally considered to be due to strong ion drag force at the center of plasma [39, 40, 41].
The observation of Coulomb crystals in fine particle plasmas was presented in this chapter. Their 3D crystal structures were fcc, fco, and bct, but bcc structure have not been observed. The latter result is due to the fact that the rearrangement from fcc to fco or bct occurs with constant particle density in horizontal planes and with interplane vertical distance constant. It is explained that the crystal structure was formed from the pile of the plain layers perpendicular to the direction of an external force with constant interplane distance being arranged by the secondary effect of the Coulomb force of particles in nearby layers to minimize the average Coulomb energy.
The structure change of Coulomb crystal during the growth of carbon particles was observed and analyzed with the use of Mie-scattering ellipsometry. The arrangement of fine particles 1.3 μm in diameter showed 3D regularity, while that of diameter 2.2 μm formed 2D close-packed crystal structure.
The spatial distribution of size of particles forming horizontal layers was analyzed by Mie-scattering ellipsometry using an image sensor. By the comparison of the determined values Ψ and Δ with calculation, particle diameter was evaluated to be 2.70, 2.74, 2.75, and 2.77 μm for particles in upper to lower layers. Its diameter was determined with accuracy as low as 0.01 μm (10 nm). Such measurement method may be useful for the analysis of phase separation phenomena, which is observed in general materials, in fine particle plasmas.
3D Coulomb crystals in fine particle plasmas can be good models of real atomic crystal and its formation and melting processes. Thus physical phenomena in solid, liquid, and gas states, which are, for example, the liquid-to-solid phase transition [46, 47], critical phenomena [48], soliton [49], chaos [50], can be simulated and analyzed on the basis of observation of behavior of individual fine particles. Such an experimental method enables the kinetic analyses of component behavior in the study of statistical properties of a system, along with the complementary uses of the computer simulations of molecular dynamics (MD) or Monte-Carlo method (MC).
In order to analyze a 3D Coulomb crystal as a model of real atomic crystal, it is preferable to make its conditions of environment close to the real crystal. Spatially isotropic force field acting on fine particles is required for the conditions. Microgravity environment can eliminate unidirectional gravity; however, the resultant force of electrostatic and ion drag forces pushes fine particles outward from the center forming the void of them [44, 45]. The fabrication of a plasma system that does not generate such a void is yet an unresolved issue and that with homogeneous conditions is under development [51, 52].
The author thanks Prof. Kunihide Tachibana, Prof. Kazuo Takahashi, Prof. Akio Sanpei, Prof. Yukio Watanabe, Prof. Noriyoshi Sato, and members of the project of Plasma Control Science Research in Kyoto Institute of Technology for useful discussions. He also thanks laboratory students of Kyoto Institute of Technology for experimental assistance. This work was partly supported by the Ministry of Education, Science, Sports and Culture of Japan.
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\n\n\n\nBook Chapters and Monographs
\n\n\n\nCorresponding authors will receive a 25% discount on their Open Access Publication Fees (OAPF) for Open Access book chapters. A 20% discount for publishing a long-form monographs, 25% for compacts and 23% for short-form monographs.
\n\nCSIC affiliated authors can also take advantage of a central Open Access fund (amounting to 10,000 EUR) to cover up to 50% of the rest of the OAPF until it expires. Effective for chapters accepted from January 1, 2020.
\n\nCorresponding authors will receive a 25% discount on their Open Access Publication Fees (OAPF) for Open Access book chapters. A 20% discount for publishing a long-form monographs, 25% for compacts and 23% for short-form monographs.
\n\nCorresponding authors will receive a 25% discount on their Open Access Publication Fees (OAPF) for Open Access book chapters. A 20% discount for publishing a long-form monographs, 25% for compacts and 23% for short-form monographs.
\n\n\n\nCorresponding authors will receive a 25% discount on their Open Access Publication Fees (OAPF) for Open Access book chapters. A 20% discount for publishing a long-form monographs, 25% for compacts and 23% for short-form monographs.
\n\nBook Chapters and Monographs
\n\nBook Chapters and Monographs
\n\nBook Chapters and Monographs
\n\n\n\nBook Chapters and Monographs
\n\nThe Claremont Colleges are pledging funds via the Knowledge Unlatched program to ensure academics can publish Open Access content more easily.
\n\nCorresponding authors will receive a 15% discount on their Open Access Publication Fees (OAPF) for Open Access book chapters or monograph publications. To use the discount you will need to verify your institutional email address. These discounts are valid from 2020 to 2022.
\n\nThe University of Massachusetts, Amherst is pledging funds via the Knowledge Unlatched program to ensure academics can publish Open Access content more easily.
\n\nCorresponding authors will receive a 10% discount on their Open Access Publication Fees (OAPF) for Open Access book chapters or monograph publications. To use the discount you will need to verify your institutional email address. These discounts are valid from 2020 to 2022.
\n\nThe University of Surrey is pledging funds via the Knowledge Unlatched program to ensure academics can publish Open Access content more easily.
\n\nCorresponding authors will receive a 10% discount on their Open Access Publication Fees (OAPF) for Open Access book chapters or monograph publications. To use the discount you will need to verify your institutional email address. These discounts are valid from 2020 to 2022.
\n\nMonographs Only
\n\n\n\nImportant: You must be a member or grantee of the above listed institutions in order to apply for their Open Access publication funds.
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Lauret",authors:[{id:"86588",title:"Prof.",name:"Jean Philippe",middleName:null,surname:"Gastellu-Etchegorry",slug:"jean-philippe-gastellu-etchegorry",fullName:"Jean Philippe Gastellu-Etchegorry"}]},{id:"28865",doi:"10.5772/30323",title:"3D Modelling from Real Data",slug:"3d-modeling-from-real-data",totalDownloads:3411,totalCrossrefCites:14,totalDimensionsCites:22,abstract:null,book:{id:"907",slug:"modeling-and-simulation-in-engineering",title:"Modeling and Simulation in Engineering",fullTitle:"Modeling and Simulation in Engineering"},signatures:"Gabriele Guidi and Fabio Remondino",authors:[{id:"81959",title:"Prof.",name:"Gabriele",middleName:null,surname:"Guidi",slug:"gabriele-guidi",fullName:"Gabriele Guidi"},{id:"89325",title:"Dr.",name:"Fabio",middleName:null,surname:"Remondino",slug:"fabio-remondino",fullName:"Fabio Remondino"}]},{id:"28868",doi:"10.5772/29744",title:"Open Source 3D Game Engines for Serious Games Modeling",slug:"open-source-3d-game-engines-for-serious-games-modeling",totalDownloads:5655,totalCrossrefCites:7,totalDimensionsCites:14,abstract:null,book:{id:"907",slug:"modeling-and-simulation-in-engineering",title:"Modeling and Simulation in Engineering",fullTitle:"Modeling and Simulation in Engineering"},signatures:"Andres Navarro, Juan Vicente Pradilla and Octavio Rios",authors:[{id:"79143",title:"Prof.",name:"Andres",middleName:null,surname:"Navarro Cadavid",slug:"andres-navarro-cadavid",fullName:"Andres Navarro Cadavid"},{id:"90462",title:"BSc.",name:"Juan Vicente",middleName:null,surname:"Pradilla",slug:"juan-vicente-pradilla",fullName:"Juan Vicente Pradilla"},{id:"90463",title:"BSc.",name:"Octavio",middleName:null,surname:"Rios",slug:"octavio-rios",fullName:"Octavio Rios"}]},{id:"28867",doi:"10.5772/31092",title:"Applications of Computational 3D–Modeling in Organismal Biology",slug:"applications-of-computational-3d-modeling-in-biological-sciences",totalDownloads:2977,totalCrossrefCites:5,totalDimensionsCites:12,abstract:null,book:{id:"907",slug:"modeling-and-simulation-in-engineering",title:"Modeling and Simulation in Engineering",fullTitle:"Modeling and Simulation in Engineering"},signatures:"Christian Laforsch, Hannes Imhof, Robert Sigl, Marcus Settles, Martin Heß and Andreas Wanninger",authors:[{id:"57406",title:"Prof.",name:"Christian",middleName:null,surname:"Laforsch",slug:"christian-laforsch",fullName:"Christian Laforsch"},{id:"114845",title:"MSc.",name:"Hannes",middleName:null,surname:"Imhof",slug:"hannes-imhof",fullName:"Hannes Imhof"},{id:"131512",title:"MSc.",name:"Robert",middleName:null,surname:"Sigl",slug:"robert-sigl",fullName:"Robert Sigl"},{id:"131514",title:"Mr.",name:"Marcus",middleName:null,surname:"Settles",slug:"marcus-settles",fullName:"Marcus Settles"},{id:"131516",title:"Prof.",name:"Martin",middleName:null,surname:"Heß",slug:"martin-hess",fullName:"Martin Heß"},{id:"131517",title:"Prof.",name:"Andreas",middleName:null,surname:"Wanninger",slug:"andreas-wanninger",fullName:"Andreas Wanninger"}]},{id:"28871",doi:"10.5772/25955",title:"Virtual Prototyping for Rapid Product Development",slug:"virtual-prototyping-for-rapid-product-development",totalDownloads:4287,totalCrossrefCites:3,totalDimensionsCites:5,abstract:null,book:{id:"907",slug:"modeling-and-simulation-in-engineering",title:"Modeling and Simulation in Engineering",fullTitle:"Modeling and Simulation in Engineering"},signatures:"S.H. Choi and H.H. Cheung",authors:[{id:"13664",title:"Dr.",name:"S.H.",middleName:null,surname:"Choi",slug:"s.h.-choi",fullName:"S.H. Choi"},{id:"13744",title:"Dr.",name:"H.H.",middleName:null,surname:"Cheung",slug:"h.h.-cheung",fullName:"H.H. Cheung"}]}],mostDownloadedChaptersLast30Days:[{id:"62639",title:"Reliability Evaluation for Mechanical Systems by Petri Nets",slug:"reliability-evaluation-for-mechanical-systems-by-petri-nets",totalDownloads:1120,totalCrossrefCites:4,totalDimensionsCites:5,abstract:"The current trend in mechanical engineering is to design mechanical systems with higher stability, reliability, availability and operability. In order to meet the requirement of high reliability for a machine, it is of great importance for designers to seek the weak links in the system and learn the state of the key subsystems, carrying out the remedial measures when necessary. Hence, behavior modeling and failure analysis are the two aspects seriously concerned in the reliability evaluation in mechanical systems. This chapter will introduce new methodologies that use the fuzzy reasoning Petri net (FRPN) models to evaluate the reliability of mechanical systems in reliability prediction, reliability apportionment and reliability analysis. Cases are proposed by analyzing a spacecraft solar array system using the proposed method. Results indicate that the Petri nets models can contribute to a higher accuracy in reliability evaluation for mechanical systems.",book:{id:"6715",slug:"petri-nets-in-science-and-engineering",title:"Petri Nets in Science and Engineering",fullTitle:"Petri Nets in Science and Engineering"},signatures:"Jianing Wu and Shaoze Yan",authors:[{id:"238979",title:"Dr.",name:"Jianing",middleName:null,surname:"Wu",slug:"jianing-wu",fullName:"Jianing Wu"}]},{id:"54600",title:"Textile Forms’ Computer Simulation Techniques",slug:"textile-forms-computer-simulation-techniques",totalDownloads:1617,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"Computer simulation techniques of textile forms already represent an important tool for textile and garment designers, since they offer numerous advantages, such as quick and simple introduction of changes while developing a model in comparison with conventional techniques. Therefore, the modeling and simulation of textile forms will always be an important issue and challenge for the researchers, since close‐to‐reality models are essential for understanding the performance and behavior of textile materials. This chapter deals with computer simulation of different textile forms. In the introductory part, it reviews the development of complex modeling and simulation techniques related to different textile forms. The main part of the chapter focuses on study of the fabric and fused panel drape by using the finite element method and on development of some representative textile forms, above all, on functional and protective clothing for persons who are sitting during performing different activities. Computer simulation techniques and scanned 3D body models in a sitting posture are used for this purpose. Engineering approaches to textile forms’ design for particular purposes, presented in this chapter, show benefits and limitations of specific 3D body scanning and computer simulation techniques and outline the future research challenges.",book:{id:"5707",slug:"computer-simulation",title:"Computer Simulation",fullTitle:"Computer Simulation"},signatures:"Andreja Rudolf, Slavica Bogović, Beti Rogina Car, Andrej Cupar,\nZoran Stjepanovič and Simona Jevšnik",authors:[{id:"180695",title:"Prof.",name:"Simona",middleName:null,surname:"Jevšnik",slug:"simona-jevsnik",fullName:"Simona Jevšnik"},{id:"181508",title:"Prof.",name:"Zoran",middleName:null,surname:"Stjepanovič",slug:"zoran-stjepanovic",fullName:"Zoran Stjepanovič"},{id:"195721",title:"Dr.",name:"Andreja",middleName:null,surname:"Rudolf",slug:"andreja-rudolf",fullName:"Andreja Rudolf"},{id:"196110",title:"Dr.",name:"Slavica",middleName:null,surname:"Bogović",slug:"slavica-bogovic",fullName:"Slavica Bogović"},{id:"196111",title:"Ph.D.",name:"Beti",middleName:null,surname:"Rogina-Car",slug:"beti-rogina-car",fullName:"Beti Rogina-Car"},{id:"196113",title:"Dr.",name:"Andrej",middleName:null,surname:"Cupar",slug:"andrej-cupar",fullName:"Andrej Cupar"}]},{id:"59877",title:"Petri Networks in the Planning of Discrete Manufacturing Processes",slug:"petri-networks-in-the-planning-of-discrete-manufacturing-processes",totalDownloads:1044,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"This chapter puts forward characteristics of selected issues of manufacturing processes planning using the Petri networks technique. It includes references to the extensive literature concerning the use of Petri networks in computer aided planning of discrete production processes. Diversity of these problems is high as it refers both to the methods of modeling and simulation of the course of manufacturing processes, the issue of optimizing these processes and production systems, representation of knowledge on production parts of equipment and integration of planning and production activities in general. The work puts forward example use of a temporary, priority Petri network for modeling and optimizing production systems and manufacturing operations as well as an example of fuzzy interference using the Petri network mechanism.",book:{id:"6715",slug:"petri-nets-in-science-and-engineering",title:"Petri Nets in Science and Engineering",fullTitle:"Petri Nets in Science and Engineering"},signatures:"Roman Stryczek",authors:[{id:"238376",title:"Ph.D.",name:"Roman",middleName:null,surname:"Stryczek",slug:"roman-stryczek",fullName:"Roman Stryczek"}]},{id:"60326",title:"Performance Analysis of Shared-Memory Bus-Based Multiprocessors Using Timed Petri Nets",slug:"performance-analysis-of-shared-memory-bus-based-multiprocessors-using-timed-petri-nets",totalDownloads:956,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"In shared-memory bus-based multiprocessors, the number of processors is often limited by the (shared) bus; when the utilization of the bus approaches 100%, processors spend an increasing amount of time waiting to get access to the bus (and shared memory) and this degrades their performance. The limitations imposed by the bus depend upon many parameters, and different parameters affect the performance in different ways. This chapter uses timed Petri nets to model shared-memory bus-based multiprocessors at the instruction execution level and shows how the performance of processors and the system are affected by different modeling parameters. Discrete-event simulation of the developed net models is used to get performance results.",book:{id:"6715",slug:"petri-nets-in-science-and-engineering",title:"Petri Nets in Science and Engineering",fullTitle:"Petri Nets in Science and Engineering"},signatures:"Wlodek M. Zuberek",authors:[{id:"146442",title:"Dr.",name:"Wlodek",middleName:null,surname:"Zuberek",slug:"wlodek-zuberek",fullName:"Wlodek Zuberek"}]},{id:"55001",title:"Computer Simulation of Bioprocess",slug:"computer-simulation-of-bioprocess",totalDownloads:1724,totalCrossrefCites:1,totalDimensionsCites:4,abstract:"Bioprocess optimization is important in order to make the bioproduction process more efficient and economic. The conventional optimization methods are costly and less efficient. On the other hand, modeling and computer simulation can reveal the mechanisms behind the phenomenon to some extent, to assist the deep analysis and efficient optimization of bioprocesses. In this chapter, modeling and computer simulation of microbial growth and metabolism kinetics, bioreactor dynamics, bioreactor feedback control will be made to show the application methods and the usefulness of modeling and computer simulation methods in optimization of the bioprocess technology.",book:{id:"5707",slug:"computer-simulation",title:"Computer Simulation",fullTitle:"Computer Simulation"},signatures:"Jianqun Lin, Ling Gao, Huibin Lin, Yilin Ren, Yutian Lin and\nJianqiang Lin",authors:[{id:"16729",title:"Dr.",name:"Jianqun",middleName:null,surname:"Lin",slug:"jianqun-lin",fullName:"Jianqun Lin"},{id:"16857",title:"Prof.",name:"Ling",middleName:null,surname:"Gao",slug:"ling-gao",fullName:"Ling Gao"},{id:"16859",title:"Dr.",name:"Jianqiang",middleName:null,surname:"Lin",slug:"jianqiang-lin",fullName:"Jianqiang Lin"},{id:"205062",title:"Mrs.",name:"Yutian",middleName:null,surname:"Lin",slug:"yutian-lin",fullName:"Yutian Lin"},{id:"205111",title:"Prof.",name:"Yilin",middleName:null,surname:"Ren",slug:"yilin-ren",fullName:"Yilin Ren"},{id:"205113",title:"Prof.",name:"Huibin",middleName:null,surname:"Lin",slug:"huibin-lin",fullName:"Huibin Lin"}]}],onlineFirstChaptersFilter:{topicId:"559",limit:6,offset:0},onlineFirstChaptersCollection:[],onlineFirstChaptersTotal:0},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:0,limit:8,total:null},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:9,numberOfPublishedChapters:90,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:104,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:32,numberOfPublishedChapters:320,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:13,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:11,numberOfPublishedChapters:141,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:8,numberOfPublishedChapters:133,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!0},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:114,numberOfOpenTopics:3,numberOfUpcomingTopics:1,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:11,numberOfPublishedChapters:107,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:19,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2753-894X",doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:7,numberOfOpenTopics:1,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!0},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:0,numberOfPublishedChapters:17,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:null,doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}},{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. The whole process of submitting an article and editing of the submitted article goes extremely smooth and fast, the number of reads and downloads of chapters is high, and the contributions are also frequently cited.",author:{id:"55578",name:"Antonio",surname:"Jurado-Navas",institutionString:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRisIQAS/Profile_Picture_1626166543950",slug:"antonio-jurado-navas",institution:{id:"720",name:"University of Malaga",country:{id:null,name:"Spain"}}}}]},series:{item:{id:"10",title:"Physiology",doi:"10.5772/intechopen.72796",issn:"2631-8261",scope:"Modern physiology requires a comprehensive understanding of the integration of tissues and organs throughout the mammalian body, including the cooperation between structure and function at the cellular and molecular levels governed by gene and protein expression. While a daunting task, learning is facilitated by identifying common and effective signaling pathways mediated by a variety of factors employed by nature to preserve and sustain homeostatic life. \r\nAs a leading example, the cellular interaction between intracellular concentration of Ca+2 increases, and changes in plasma membrane potential is integral for coordinating blood flow, governing the exocytosis of neurotransmitters, and modulating gene expression and cell effector secretory functions. 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His primary area of interest is physiology and pathophysiology of the gastrointestinal (GI) tract, with the major focus on the mechanism of GI mucosal defense, protection, and ulcer healing. He was a postdoctoral NIH fellow at the University of California and the Gastroenterology VA Medical Center, Irvine, Long Beach, CA, USA, and at the Gastroenterology Clinics Erlangen-Nuremberg and Munster in Germany. He has published 290 original articles in some of the most prestigious scientific journals and seven book chapters on the pathophysiology of the GI tract, gastroprotection, ulcer healing, drug therapy of peptic ulcers, hormonal regulation of the gut, and inflammatory bowel disease.",institutionString:null,institution:{name:"Jagiellonian University",institutionURL:null,country:{name:"Poland"}}},editorTwo:null,editorThree:null},subseries:{paginationCount:4,paginationItems:[{id:"10",title:"Animal Physiology",coverUrl:"https://cdn.intechopen.com/series_topics/covers/10.jpg",isOpenForSubmission:!0,editor:{id:"202192",title:"Dr.",name:"Catrin",middleName:null,surname:"Rutland",slug:"catrin-rutland",fullName:"Catrin Rutland",profilePictureURL:"https://mts.intechopen.com/storage/users/202192/images/system/202192.png",biography:"Catrin Rutland is an Associate Professor of Anatomy and Developmental Genetics at the University of Nottingham, UK. She obtained a BSc from the University of Derby, England, a master’s degree from Technische Universität München, Germany, and a Ph.D. from the University of Nottingham. She undertook a post-doctoral research fellowship in the School of Medicine before accepting tenure in Veterinary Medicine and Science. Dr. Rutland also obtained an MMedSci (Medical Education) and a Postgraduate Certificate in Higher Education (PGCHE). She is the author of more than sixty peer-reviewed journal articles, twelve books/book chapters, and more than 100 research abstracts in cardiovascular biology and oncology. She is a board member of the European Association of Veterinary Anatomists, Fellow of the Anatomical Society, and Senior Fellow of the Higher Education Academy. Dr. Rutland has also written popular science books for the public. https://orcid.org/0000-0002-2009-4898. www.nottingham.ac.uk/vet/people/catrin.rutland",institutionString:null,institution:{name:"University of Nottingham",institutionURL:null,country:{name:"United Kingdom"}}},editorTwo:null,editorThree:null},{id:"11",title:"Cell Physiology",coverUrl:"https://cdn.intechopen.com/series_topics/covers/11.jpg",isOpenForSubmission:!0,editor:{id:"133493",title:"Prof.",name:"Angel",middleName:null,surname:"Catala",slug:"angel-catala",fullName:"Angel Catala",profilePictureURL:"https://mts.intechopen.com/storage/users/133493/images/3091_n.jpg",biography:"Prof. Dr. Angel Catalá \r\nShort Biography Angel Catalá was born in Rodeo (San Juan, Argentina). He studied \r\nchemistry at the Universidad Nacional de La Plata, Argentina, where received aPh.D. degree in chemistry (Biological Branch) in 1965. From\r\n1964 to 1974, he worked as Assistant in Biochemistry at the School of MedicineUniversidad Nacional de La Plata, Argentina. From 1974 to 1976, he was a Fellowof the National Institutes of Health (NIH) at the University of Connecticut, Health Center, USA. From 1985 to 2004, he served as a Full Professor oBiochemistry at the Universidad Nacional de La Plata, Argentina. He is Member ofthe National Research Council (CONICET), Argentina, and Argentine Society foBiochemistry and Molecular Biology (SAIB). His laboratory has been interested for manyears in the lipid peroxidation of biological membranes from various tissues and different species. Professor Catalá has directed twelve doctoral theses, publishedover 100 papers in peer reviewed journals, several chapters in books andtwelve edited books. Angel Catalá received awards at the 40th InternationaConference Biochemistry of Lipids 1999: Dijon (France). W inner of the Bimbo PanAmerican Nutrition, Food Science and Technology Award 2006 and 2012, South AmericaHuman Nutrition, Professional Category. 2006 award in pharmacology, Bernardo\r\nHoussay, in recognition of his meritorious works of research. 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The motor of the society is the industry and the research of this topic has to be empowered in order to increase and improve the quality of our lives.",coverUrl:"https://cdn.intechopen.com/series_topics/covers/22.jpg",hasOnlineFirst:!0,hasPublishedBooks:!0,annualVolume:11418,editor:{id:"27170",title:"Prof.",name:"Carlos",middleName:"M.",surname:"Travieso-Gonzalez",slug:"carlos-travieso-gonzalez",fullName:"Carlos Travieso-Gonzalez",profilePictureURL:"https://mts.intechopen.com/storage/users/27170/images/system/27170.jpeg",biography:"Carlos M. Travieso-González received his MSc degree in Telecommunication Engineering at Polytechnic University of Catalonia (UPC), Spain in 1997, and his Ph.D. degree in 2002 at the University of Las Palmas de Gran Canaria (ULPGC-Spain). He is a full professor of signal processing and pattern recognition and is head of the Signals and Communications Department at ULPGC, teaching from 2001 on subjects on signal processing and learning theory. His research lines are biometrics, biomedical signals and images, data mining, classification system, signal and image processing, machine learning, and environmental intelligence. He has researched in 52 international and Spanish research projects, some of them as head researcher. He is co-author of 4 books, co-editor of 27 proceedings books, guest editor for 8 JCR-ISI international journals, and up to 24 book chapters. He has over 450 papers published in international journals and conferences (81 of them indexed on JCR – ISI - Web of Science). He has published seven patents in the Spanish Patent and Trademark Office. He has been a supervisor on 8 Ph.D. theses (11 more are under supervision), and 130 master theses. He is the founder of The IEEE IWOBI conference series and the president of its Steering Committee, as well as the founder of both the InnoEducaTIC and APPIS conference series. He is an evaluator of project proposals for the European Union (H2020), Medical Research Council (MRC, UK), Spanish Government (ANECA, Spain), Research National Agency (ANR, France), DAAD (Germany), Argentinian Government, and the Colombian Institutions. He has been a reviewer in different indexed international journals (<70) and conferences (<250) since 2001. He has been a member of the IASTED Technical Committee on Image Processing from 2007 and a member of the IASTED Technical Committee on Artificial Intelligence and Expert Systems from 2011. \n\nHe has held the general chair position for the following: ACM-APPIS (2020, 2021), IEEE-IWOBI (2019, 2020 and 2020), A PPIS (2018, 2019), IEEE-IWOBI (2014, 2015, 2017, 2018), InnoEducaTIC (2014, 2017), IEEE-INES (2013), NoLISP (2011), JRBP (2012), and IEEE-ICCST (2005)\n\nHe is an associate editor of the Computational Intelligence and Neuroscience Journal (Hindawi – Q2 JCR-ISI). He was vice dean from 2004 to 2010 in the Higher Technical School of Telecommunication Engineers at ULPGC and the vice dean of Graduate and Postgraduate Studies from March 2013 to November 2017. 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