\r\n\tGoverning equations of the flows and heat transfer with EHD consist of the Navier–Stokes equations, thermal effects, and additional EHD forces. Due to the complex nature of EHD, only a limited number of publications concerning modeling of the effects of EHD on laminar flows, without numerical solutions, can be identified.
",isbn:null,printIsbn:"979-953-307-X-X",pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,isSalesforceBook:!1,isNomenclature:!1,hash:"a555a6ba490d37aed450e899a08b13ab",bookSignature:"Dr. Mohsen Sheikholeslami Kandelousi",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/8914.jpg",keywords:"Electric Field, Nanofluid, Electrode Arrangements, Ferrofluid, Transportation ,Heat Transfer, Joule Heating, Lorentz Forces, Kelvin Forces, Porous Media, Coulomb Forces, Natural Convection, Forced Convection, Mixed Convection, Scaling Analysis,Enhanced Heat Transfer, Semi Analytical Methods, Numerical Simulation",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"March 8th 2019",dateEndSecondStepPublish:"March 29th 2019",dateEndThirdStepPublish:"May 28th 2019",dateEndFourthStepPublish:"August 16th 2019",dateEndFifthStepPublish:"October 15th 2019",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"3 years",secondStepPassed:!0,areRegistrationsClosed:!0,currentStepOfPublishingProcess:5,editedByType:null,kuFlag:!1,biosketch:null,coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"185811",title:"Dr.",name:"Mohsen",middleName:null,surname:"Sheikholeslami Kandelousi",slug:"mohsen-sheikholeslami-kandelousi",fullName:"Mohsen Sheikholeslami Kandelousi",profilePictureURL:"https://mts.intechopen.com/storage/users/185811/images/system/185811.jpeg",biography:"Dr. Mohsen Sheikholeslami works at the Babol Noshirvani University of Technology’s Department of Mechanical Engineering in\nIran. He is Head of the Renewable energy systems and nanofluid\napplications in heat transfer Laboratory at Babol Noshirvani University of Technology. His research interests are nanofluid, CFD,\nsimulation, mesoscopic modeling, nonlinear science, magnetohydrodynamic, ferrohydrodynamic, electrohydrodynamic, and heat\nexchangers. He has written several papers and books in various fields of mechanical\nengineering. He is the first scientist to develop a new numerical method (CVFEM)\nand he published the reference book with title: “Application of Control Volume\nBased Finite Element Method (CVFEM) for Nanofluid Flow and Heat Transfer”. He\nis also the first author of the following books: “Applications of Nanofluid for Heat\nTransfer Enhancement”, “Application of semi analytical methods for nanofluid flow\nand heat transfer”, “Hydrothermal Analysis in Engineering Using Control Volume\nFinite Element Method”, and “External Magnetic Field Effects on Hydrothermal\nTreatment of Nanofluid”, which are published in ELSEVIER. According to the\nreports of Thomson Reuters (Clarivate Analytics), he has been selected as a Web of\nScience Highly Cited Researcher (Top 0.01%) in 2016, 2017, and 2018.",institutionString:"Babol Noshirvani University of Technology",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"6",institution:{name:"Babol Noshirvani University of Technology",institutionURL:null,country:{name:"Iran"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"11",title:"Engineering",slug:"engineering"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"288104",firstName:"Ivana",lastName:"Spajic",middleName:null,title:"Ms.",imageUrl:"https://mts.intechopen.com/storage/users/288104/images/8497_n.jpg",email:"ivana.s@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review, to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. Whether that be identifying an exceptional author and proposing an editorship collaboration, or contacting researchers who would like the opportunity to work with IntechOpen, I establish and help manage author and editor acquisition and contact."}},relatedBooks:[{type:"book",id:"6514",title:"Microfluidics and Nanofluidics",subtitle:null,isOpenForSubmission:!1,hash:"4ec06fd827f4dc0d3d7653eda88662de",slug:"microfluidics-and-nanofluidics",bookSignature:"Mohsen Sheikholeslami Kandelousi",coverURL:"https://cdn.intechopen.com/books/images_new/6514.jpg",editedByType:"Edited by",editors:[{id:"185811",title:"Dr.",name:"Mohsen",surname:"Sheikholeslami Kandelousi",slug:"mohsen-sheikholeslami-kandelousi",fullName:"Mohsen Sheikholeslami Kandelousi"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"5406",title:"Nanofluid Heat and Mass Transfer in Engineering Problems",subtitle:null,isOpenForSubmission:!1,hash:"d47e243d462589591986c11b07212df8",slug:"nanofluid-heat-and-mass-transfer-in-engineering-problems",bookSignature:"Mohsen Sheikholeslami Kandelousi",coverURL:"https://cdn.intechopen.com/books/images_new/5406.jpg",editedByType:"Edited by",editors:[{id:"185811",title:"Dr.",name:"Mohsen",surname:"Sheikholeslami Kandelousi",slug:"mohsen-sheikholeslami-kandelousi",fullName:"Mohsen Sheikholeslami Kandelousi"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6807",title:"HVAC System",subtitle:null,isOpenForSubmission:!1,hash:"4805829f41fa799b707e4d442eac16da",slug:"hvac-system",bookSignature:"Mohsen Sheikholeslami Kandelousi",coverURL:"https://cdn.intechopen.com/books/images_new/6807.jpg",editedByType:"Edited by",editors:[{id:"185811",title:"Dr.",name:"Mohsen",surname:"Sheikholeslami Kandelousi",slug:"mohsen-sheikholeslami-kandelousi",fullName:"Mohsen Sheikholeslami Kandelousi"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6362",title:"Electric Field",subtitle:null,isOpenForSubmission:!1,hash:"70b535bf877d17b46ddd1678574792a0",slug:"electric-field",bookSignature:"Mohsen Sheikholeslami Kandelousi",coverURL:"https://cdn.intechopen.com/books/images_new/6362.jpg",editedByType:"Edited by",editors:[{id:"185811",title:"Dr.",name:"Mohsen",surname:"Sheikholeslami Kandelousi",slug:"mohsen-sheikholeslami-kandelousi",fullName:"Mohsen Sheikholeslami Kandelousi"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7487",title:"Thermal Energy Battery with Nano-enhanced PCM",subtitle:null,isOpenForSubmission:!1,hash:"a917c6664886ab78f757aeb59f45635d",slug:"thermal-energy-battery-with-nano-enhanced-pcm",bookSignature:"Mohsen Sheikholeslami Kandelousi",coverURL:"https://cdn.intechopen.com/books/images_new/7487.jpg",editedByType:"Edited by",editors:[{id:"185811",title:"Dr.",name:"Mohsen",surname:"Sheikholeslami Kandelousi",slug:"mohsen-sheikholeslami-kandelousi",fullName:"Mohsen Sheikholeslami Kandelousi"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7774",title:"Nanofluid Flow in Porous Media",subtitle:null,isOpenForSubmission:!1,hash:"694361f15eb61a1b21ff01c6cd96f59a",slug:"nanofluid-flow-in-porous-media",bookSignature:"Mohsen Sheikholeslami Kandelousi, Sadia Ameen, M. 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Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"61016",title:"Minor and Trace Elements in Whole Blood, Tissues, Proteins and Immunoglobulins of Mammals",doi:"10.5772/intechopen.75939",slug:"minor-and-trace-elements-in-whole-blood-tissues-proteins-and-immunoglobulins-of-mammals",body:'\n
\n
1. Introduction
\n
Minor and trace elements including metals play important roles in many systems, normal and pathologic processes. Nowadays, there are many methods for elemental analysis of different biological samples. Atomic absorption spectrometry (AAS), inductively coupled plasma atomic emission spectrometry (ICP-AES) and mass spectrometry (ICP-MS) are usually used for analysis of blood and animal tissues [1, 2]. Generally, these methods require matrix destruction with concentrated acids. Using microwave-assisted wet acid digestion with temperature control and elevated pressure allows reducing the time of sample digestion and risk of element losses. For direct ICP-MS analysis of whole blood and serum, dilution with alkaline solutions containing EDTA, ammonia and Triton X-100 was also employed to lyse the blood cells and prevent blood clotting [3, 4]. To improve the analytical capabilities of the methods applied, the new analytical techniques and reagents are being developed. A collision/reaction cell technology allowed the removal of polyatomic interferences and extended the capabilities of ICP-MS for trace element determination [5]. To decrease limits of detection (LODs) of trace elements, different ways of their preconcentration were offered. For determining the low concentration of Pb in blood serum by flame AAS, Barbosa with coauthors [6] used oxidized carbon nanotubes covered with bovine serum albumin layers; preconcentration was performed in untreated blood serum and allowed getting LOD of Pb at the level of 2 μg/L. Mortada with coauthors [7] suggested hydroxyapatite nanorods prepared from recycled eggshell for solid phase extraction of Pb, Cu and Zn from solutions of different biological samples followed by AAS analysis. In spite of undeniable progress in developing the above-mentioned methods, the analysis of biological samples using solid sampling is very attractive as the analytical procedure is simple and risk of contamination and analyte losses is improbable. To adapt the above methods for direct analysis of solid biological samples, laser ablation (LA) and electrothermal vaporization (ETV) were applied. LA-ICP-MS has got significant attention over the last decade for the analysis of biological samples [8]. It has been mainly applied to produce images of element distributions in human and animal tissues, which is of great scientific interest [9]. The main challenge of facing the LA-ICP-MS application is fully quantitative analysis requiring complex strategies for producing reliable calibrating materials. Lack of certified reference materials (CRMs) with different biological matrices and complications of preparing matrix-matched calibration samples [10] often make it difficult to carry out quantitative multi-elemental analysis of different biological tissues. This problem is well known for ETV-ICP-AES, X-ray fluorescence and laser-induced breakdown spectrometry (LIBS).
\n
In recent times, LIBS has been applied as a screening tool for trace element bio-imaging in human and animal organs. To detect Wilson’s disease, the study of Cu distribution in human liver was carried out [11]. A low-cost approach allows the quick detection of pathological accumulation of Cu in the affected organs. The LIBS mapping of the mice kidney slices after injection of a solution containing Gd-based nanoparticles was performed [12]. Each of the above methods has both advantages and disadvantages, and the choice of the analytical method depends on the sample nature, analytical tasks and availability of appropriate device and calibration samples. In the present work, two-jet plasma atomic emission spectrometry (TJP-AES) was used for analysis of different biological samples.
\n
\n
\n
2. Two-jet plasma atomic emission spectrometry
\n
\n
2.1. A two-jet plasmatron
\n
The TJP was developed by Zheenbaev and Engel’sht in the USSR (Kyrgyzstan Institute of Physics) in the mid-1970s [13]. It is a direct current (dc) plasma that differs from dc plasmas described [14, 15] and an ICP by a high power, which allows analysis of powdered samples without sample dissolution. Although the TJP appeared approximately at the same time as an ICP, TJP-AES was not generally recognized since only a few copies of the plasmatron were produced; it has not been modernized for decades. Nowadays, TJP-AES is experiencing a new stage in its development because a new modern plasmatron was designed at “VMK-Optoelektronika” (Russia). A photograph of the plasma torch and scheme of electrode unit are presented in Figure 1.
\n
Figure 1.
(A) Plasma torch; (B) electrode unit and analytical regions of the plasma flow: 1, before the jet confluence; 2, after the jet confluence.
\n
Argon plasma jets are generated in non-consumable electrodes (copper anode and tungsten cathode); they join at the output to form plasma discharge. The argon consumption does not usually exceed 5 l/min. The power supply of the plasma generator, gas flow and automatic sample introduction systems are computer controlled. A TJP power can be fixed from 5 to 12 kW by varying the current strength in the range of 40–100 A. Current fluctuation does not exceed 1%. A new plasmatron is equipped with a concave diffraction grating (2400 lines/mm) and two multicrystal photodiode arrays allowing spectra to be measured in two spectral ranges: 190–350 and 390–450 nm.
\n
To transfer powders into the plasma, a powder introduction device was developed. The sample is placed in a Plexiglas beaker, inserted into the device and roiled with blast waves produced by a spark between zirconium electrodes over the surface of the powder. An aerosol obtained is delivered into the plasma with a carrier gas. The device allows the introduction of both small and large amount of the samples (5–500 mg).
\n
\n
\n
2.2. Analytical regions
\n
There are two analytical regions in the TJP – before and after the jet confluence (Figure 1). The region of the confluence is not used for analysis due to high background emission. A study of behavior of a wide range of elements in the plasma has shown that the highest ratio of the analytical line intensity to the background one (Il/Ib) realizes in the region before the jet confluence both for atomic and ionic lines. As an example, the distribution of Il/Ib for analytical lines Mn I 280.11, Al I 308.22; Cd I 228.80 and Fe I 302.06 along the plasma flow is shown in Figure 2.
\n
Figure 2.
Il/Ib distribution along the plasma flow for Mn I 280.11, Al I 308.22; Cd I 228.80 and Fe I 302.06 lines: region (A) before the jet confluence, (B) after the jet confluence; 0 – point in the region of the confluence.
\n
Analytical lines of Ag and Zn registered in the region before and after the jet confluence are shown in Figure 3.
\n
Figure 3.
Analytical lines Ag I 328.07 (0.05 μg/g), Zn I 213.86 (0.5 μg/g) and Zn II 206.20 (1 μg/g) registered in the region (a) before the jet confluence and (b) after the jet confluence [16].
\n
As it is seen, the analytical signals are at the level of background fluctuations in the region after the jet confluence, but they are considerably higher than background in the region before one, which testifies to better LODs of elements in this region. One of the problems arising in the TJP-AES analysis of powdered samples is their incomplete evaporation which can lead to a systematic underestimation of the analysis results. The short residence time of the sample in the plasma is one of the reasons for the partial sample evaporation. In addition, evaporation efficiency depends on chemical composition, structure and particle size of powdered samples [17]. To demonstrate evaporation efficiency in the analytical zones of the TJP, silicon carbide (SiC) powders with an average particle size of 1, 3, 7.5, 17, 22 and 36 μm were used. SiC is a heat-resistant material having a high hardness close to the hardness of diamond. SiC evaporation was controlled by the intensity of weak Si I 212.30 line. To avoid signal self-absorption, the powders were diluted with graphite 100 times. The dependence of Si I 212.30 line intensity on the particle size is given in Figure 4.
\n
Figure 4.
Dependence of Si I 212.30 line intensity on the particle size of SiC in the region (A) before the jet confluence (B) after the jet confluence.
\n
The different behavior of silicon line is observed in the regions investigated. In the region before the jet confluence, there is an increase in the silicon line intensity along with a decrease in the particle size, the smaller particle size and the better evaporation efficiency. However, the complete sample evaporation does not occur even at a particle size of 3 μm. In the region after the jet confluence, the maximum intensity is achieved even at a particle size of 17 μm. The decrease in intensity at smaller particles seems to occur due to the introduction of light particles into the region after the confluence is complicated by a resistance of the consistent plasma jets. Nevertheless, the effect of particle size on evaporation efficiency is considerably weaker in the region after the jet confluence. Thus, the region before the jet confluence provides lower detection limits of elements than the region after one, but evaporation efficiency is better in the region after the jet confluence. Therefore, the choice of the analytical region depends on an analytical task and sample nature.
\n
\n
\n
2.3. Excitation mechanisms
\n
To solve the problems appearing in the analysis, it is important to have an idea about the processes occurring in the plasma. The probable mechanisms of an atom and ion excitation in the TJP were investigated [16, 18]. The Boltzmann distribution of excited energy levels for Fe atoms and singly ionized ions was found to take place in both analytical regions, which indicates the predominant excitation of atoms and ions by electron impact. Excitation temperature of Fe atoms and ions, Tatom and Tion, respectively, was measured along the plasma flow. In the optimal observation zone of the region before the jet confluence Tatom = 6000 K and Tion = 7900 K, and in the observation zone of the region after one Tatom = 7060 K and Tion = 8050 K. For atomic and ionic lines, the temperature deviation did not exceed 100 K in the optimal observation zones and was about 250 K near the jet confluence. The considerable difference in Tatom and Tion points at the departure from local thermodynamic equilibrium (LTE) in the plasma. The difference is 1900 K for the region before the jet confluence and 990 K for the region after one, which indicates that the region before the jet confluence is more non-equilibrium than the region after one. The disturbance of LTE in the plasma was shown to be due to metastable argon participation in atom ionization.
\n
\n
\n
\n
3. Application of TJP-AES
\n
Originally, the TJP was intended for direct analysis of sparingly soluble geological samples [19], which considerably reduced the analysis time and element losses as compared with wet acid digestion. The spectra were registered in the analytical region after the jet confluence (Figure 1), and calibration samples similar to the analyzed ones were used. The region before the jet confluence was not practically used for analysis.
\n
\n
3.1. Analysis of high purity substances
\n
The region before the jet confluence turned out to be suitable for analysis of high purity substances both by direct technique and after matrix separation. The TJP-AES techniques for analysis of gallium [20], indium and indium oxide [21] and tellurium dioxide [22] were developed. The direct techniques allow determination of about 30 elements using appropriate dilution of the sample with a spectroscopic buffer (graphite powder containing 15 wt.% NaCl) and unified calibration samples based on graphite powder with addition of 15 wt.% NaCl. As it was shown earlier, NaCl addition increases analytical line intensities and suppresses effects of a mineral matrix [23]. CRMs of graphite powder with different combinations of impurities are commercially available (Ural Federal University, Russia); in addition, preparing the reference sample with given element concentration in graphite is not a difficult task. Analysis of the above substances was carried out at the optimal conditions chosen for multi-elemental analysis of graphite powder (Table 1). Calibration curves (lgI-lgC) obtained for Cd and Hg in graphite powder are presented in Figure 5.
\n
\n
\n
\n\n
\n
Parameter
\n
Value
\n
\n\n\n
\n
Current strength
\n
85 A
\n
\n
\n
Voltage
\n
120 V
\n
\n
\n
Plasma gas flow
\n
4 l/min
\n
\n
\n
Carrier gas flow
\n
0.85 l/min
\n
\n
\n
Angle between the jets
\n
60○
\n
\n
\n
Observation zone
\n
4–5 mm lower than the point of the confluence
\n
\n\n
Table 1.
Working conditions of the two-jet plasma.
\n
Figure 5.
Calibration curves for analytical lines Cd I 228.80 and Hg I 253.65 (C μg/g) [24].
\n
A degree of sample dilution depends on the sample nature; a fourfold dilution is needed for analysis of indium oxide and gallium, and a twofold dilution is quite enough for analysis of tellurium dioxide. The preconcentration of impurities in gallium and indium was accomplished by matrix separation in the form of chlorides; tellurium dioxide was previously reduced by hydrogen to metal, and the preconcentration was performed by vacuum distillation of tellurium. The impurity concentrates contained a high concentration of matrix elements since incomplete matrix separation was applied to avoid the loss of a number of important impurities; they were analyzed by the same way as in the direct techniques. LODs of elements were at the level of 0.01–1 and 0.001–0.1 μg/g for direct analysis and after matrix separation, respectively. The possibility of analysis of such a different substances using unified calibration samples points at comparatively weak matrix affects this excitation source. For comparison, using such an approach for a dc arc with sample evaporation from a crater of graphite electrode requires a 100-, 50-, and 25-fold dilution of gallium, indium and tellurium oxides, respectively, which lead to worsening LODs of elements by more than an order of magnitude. Recently, the similar approach was used for analysis of different soils [24]. In spite of their complex and variable matrix composition, TJP-AES allowed direct determination of As, B, Cd, Cu, Hg and P after a twofold and Be, Co, Cr, Ga, Nb, Pb and Zn after a 10-fold dilution with a spectroscopic buffer.
\n
\n
\n
3.2. Analysis of biological samples
\n
Solid sampling, with little or no chemical pretreatment, in the analysis of biological samples seems very attractive. The possibility of TJP-AES for direct analysis of biological samples using the same unified approach as for inorganic materials was investigated. First, starch was used to study the organic matrix influence on analytical signals of elements in the TJP [25]. It was found that the presence of 10 wt.% starch in graphite powder with introduced impurities did not affect analytical line intensities of elements while the decrease in intensities by a factor 2–5 was observed in a graphite dc arc. In the TJP, the effect was not observed even in the presence of 20 wt.% starch for many atomic lines of elements. The considerable decrease in intensities in a dc arc is due to a vigorous reaction of starch with air oxygen in an arc discharge with the release of gaseous products, which results in decreasing the residence time of the sample in the plasma. Although the TJP is an open system too, the oxidative reaction occurs less violently than in a dc arc since carrier argon partially displaces air from the excitation zone, and the sample is gradually introduced into the plasma. In addition, gaseous products seem to be retained in the excitation zone by argon flows. The experiments with starch gave hope to get positive results for more complex organic matrices.
\n
\n
3.2.1. Analysis of animal organs
\n
The unified approach mentioned above was tested for animal organs, dried and finely powdered. Dry animal organs contain more than 50 wt.% proteins as well as fats, carbohydrates and others. The effect of such a complex matrix on the analytical signal of elements was studied by the analysis of a spiked sample based on graphite with the addition of 15 wt.% NaCl and 10 wt.% rat liver; concentration of elements introduced was 2.5 μg/g [26]. The analysis was carried out using calibration samples based on graphite powder with addition 15 wt.% NaCl; the spectra were observed in the region before the jet confluence. The analysis results are given in Table 2; the satisfactory recoveries were obtained for all investigated elements. Since the liver contained Co, Mn, Mo and Zn, the blank sample was prepared to estimate correctly the recovery of these elements. On the basis of the results obtained, a 10-fold dilution of powdered animal organs with buffer and calibration based on graphite powder were suggested for direct analysis of animal organs. To validate the technique, the results of direct analysis of bovine liver were compared with the results obtained after sample carbonization (500°C, 5 min) and autoclave digestion in a mixture of nitric acid and hydrogen peroxide [25]. The carbonized sample was analyzed at the conditions chosen for direct analysis. The solution obtained after acid digestion evaporated on graphite powder, diluted with buffer and analyzed by the same way. As it is seen from Table 3, the results of Al, Ca, Cu, Fe, Mg, Mn, Mo, P, Si and Zn satisfactory agree with each other. Only for Fe, Mn and Mo, the results obtained after autoclave digestion are lower than the results of the direct technique, which is likely to be due to their partial loss. The LODs of elements provided by the direct technique are at the level 0.1–10 μg/g, and they are lower by approximately one order of magnitude after carbonization. The use of the carbonization procedure allowed determining the low concentrations of Ag, Cd, Co, Cr, Pb and Ni in the liver.
Determination of elements in a spiked graphite powder containing 10% rat liver [26].
Mean ± (95% confidence interval), n = 4.
Value obtained without spike addition.
\n
\n
\n
\n
\n
\n
\n\n
\n
B
\n
249.77
\n
6.2 ± 2.0
\n
2.7 ± 0.5
\n
5.5 ± 1.0
\n
\n
\n
Ca
\n
317.93
\n
130 ± 20
\n
110 ± 30
\n
130 ± 40
\n
\n
\n
Cd
\n
228.80
\n
<0.4
\n
0.17 ± 0.03
\n
0.25 ± 0.10
\n
\n
\n
Co
\n
345.35
\n
<1
\n
0.31 ± 0.05
\n
—
\n
\n
\n
Cu
\n
324.75
\n
12 ± 2
\n
12 ± 4
\n
10 ± 0.2
\n
\n
\n
Cr
\n
283.56
\n
<1
\n
0.2 ± 0.04
\n
0.24 ± 0.06
\n
\n
\n
Fe
\n
302.06
\n
250 ± 40
\n
210 ± 70
\n
150 ± 30
\n
\n
\n
Mg
\n
277.98
\n
470 ± 80
\n
460 ± 120
\n
420 ± 90
\n
\n
\n
Mn
\n
279.83
\n
16 ± 2
\n
13 ± 3
\n
9.5 ± 2.4
\n
\n
\n
Mo
\n
317.03
\n
18 ± 3
\n
20 ± 5
\n
9.0 ± 2.3
\n
\n
\n
Ni
\n
305.08
\n
<1
\n
0.6 ± 0.1
\n
0.77 ± 0.25
\n
\n
\n
P, wt.%
\n
214.91
\n
0.99 ± 0.08
\n
0.85 ± 0.25
\n
0.76 ± 0.11
\n
\n
\n
Pb
\n
283.31
\n
<1
\n
0.40 ± 0.07
\n
0.40 ±0.11
\n
\n
\n
Si
\n
288.16
\n
22 ± 8
\n
31 ± 14
\n
22 ± 11
\n
\n
\n
Zn
\n
213.86
\n
62 ± 15
\n
55 ± 10
\n
60 ± 8
\n
\n\n
Table 3.
Results of the TJP-AES analysis of bovine liver (μg/g) [25].
95% confidence interval.
\n
The technique suggested is very suitable for analysis of dry internals such as liver, kidney and spleen which are easily ground in a Plexiglas mortar to a powder with the particle size of 20–30 μm. However, the direct analysis of bovine and pork muscle CRMs and rat brain was found to provide the understated results [26]. These tissues are more thermostable than liver and have the particles of more than 100 μm. Flexible fibers of muscles and plastic consistence of brain make it difficult to obtain a powder with small particles. Incomplete evaporation of the samples is the most probable reason for the result underestimations. This problem was overcome by decreasing the consumption of a carrier gas, which increases the residence time of the sample in the excitation zone and concentration of air oxygen participating in organic matrix decomposition. It should be noted that the carbonization conditions are not all-purpose and depend on a kind of tissues. For brain and muscle tissues, the time of carbonization was increased up to 30 min. Thus, in spite of the unified approach to the analysis of organs, peculiarities of different tissues should be taken into account. For direct analysis, 5–10 mg of powdered sample is quite enough; for carbonization procedure, 50–100 mg of the sample is needed. Note that the ICP-AES and ICP-MS techniques with wet acid digestion of organs usually require 100–250 mg of the sample which is not always available. The relative standard deviation of the analysis results of animal organs usually is in the range of 3–12%.
\n
\n
\n
3.2.2. Analysis of whole blood
\n
The problem of availability of biological samples in ample quantity is particularly acute in experiments with living experimental animals (such as mice or rats). Blood is the main subject of investigation for living organisms. Whole blood quickly changes over time due to fast clotting, which troubles the sample cutting. The effect of anticoagulants also lasts a limited time. Therefore, for continuous biomedical experiments, freeze-dried whole blood which can be kept for a long time at normal conditions is very convenient. For determining the main essential elements (Fe, P, Ca, Mg, Zn and Cu) in freeze-dried whole blood, the direct technique developed for animal organs was applied [27]. To confirm the possibility of such an approach for blood analysis, the direct analysis of CRM of freeze-dried bovine blood (IAEA A-13) was carried out, and good agreement of the results with the certified values was obtained. In addition, the results of analysis of human and rat freeze-dried whole blood obtained using the different sample preparation procedures were compared (Table 4). The direct technique results satisfactory agreed with the results obtained after carbonization (400°C, 15 min) and autoclave digestion, which confirms the possibility of the unified approach for blood analysis. Simple sample preparation (dilution with buffer) and the possibility of analysis of small amount samples, 5–10 mg of blood powder (approximately 20–50 μL liquid blood), are of practical importance. For analysis of whole liquid blood, blood aliquots evaporated on graphite powder under an IR lamp and then carbonized at 400°C for 15 min. The remainder was ground in a mortar and analyzed as in the direct analysis. The techniques suggested were validated for analysis of both freeze-dried and liquid blood serum and plasma.
Concentration of elements in the freeze-dried blood samples (μg/g) [27].
95% confidence interval, n = 4.
\n
\n
\n
3.2.3. Analysis of bone
\n
Bone is a highly mineralized mobile tissue which accumulates inorganic substances and diffuses them as the need arises. It contains 25 and 65 wt.% organic and inorganic substances, respectively, and 10 wt.% water. Collagen and calcium hydroxyapatite Ca10(PO4)6(OH)2 are the main components of bone. Some elements are predominantly in the organic phase, and others are in the mineral phase of bone. It was found that a fourfold dilution of dry powdered bone was quite enough for element determination by the unified direct technique [28]. The results of “added-found” experiment and comparison of the direct technique results with the results of ICP-AES after wet acid digestion of the sample validated the technique. However, underestimating the Ba, Mg, and Sr concentration was obtained. These elements are strongly bound with calcium hydroxyapatite, and its incomplete evaporation may lead to their understated concentrations. This effect took place even at a mean particle size of 30 μm. The strongest decrease in concentration was observed for Sr replacing Ca in hydroxyapatite. It is well known that strontium rachitis develops in the regions with a high content of radioactive Sr due to the formation of high concentration of strontium hydroxyapatite in bone, which results in the fragility of people and animal teeth and bones. Pretreatment of the samples with nitric acid followed by heating at 300°C or decrease of the consumption of carrier argon as in the case of brain and muscle tissues allowed getting valid results for Ba, Mg, and Sr. The satisfactory results obtained for other elements by direct technique point at their fractional volatilization from the particles in the plasma. These elements seem to be bound with the organic portion of bone or weakly bound with calcium hydroxyapatite. LODs of a number of elements in bone are given in Table 5.
\n
\n
\n
\n
\n\n
\n
Element
\n
λ (nm)
\n
LOD (μg/g)
\n
\n\n\n
\n
Ag
\n
328.07
\n
0.1
\n
\n
\n
Bi
\n
306.77
\n
1.2
\n
\n
\n
Cd
\n
228.80
\n
0.5
\n
\n
\n
Co
\n
345.35
\n
1.2
\n
\n
\n
Cr
\n
283.56
\n
0.3
\n
\n
\n
Cu
\n
324.75
\n
0.2
\n
\n
\n
Fe
\n
296.68
\n
2.0
\n
\n
\n
Ga
\n
294.36
\n
0.3
\n
\n
\n
Mn
\n
260.57
\n
0.5
\n
\n
\n
Mo
\n
313.26
\n
0.7
\n
\n
\n
Ni
\n
305.08
\n
0.6
\n
\n
\n
Pb
\n
283.31
\n
1.1
\n
\n
\n
Sn
\n
284.00
\n
0.8
\n
\n
\n
Zn
\n
213.86
\n
1.0
\n
\n\n
Table 5.
Limits of detection (LOD) of elements in bone [28].
\n
Thus, on the example of different animal organs, whole blood and bone the possibility of TJP-AES for realizing the simple analytical techniques was shown. Solid sampling, unified calibration samples, the possibility of analysis of small amount samples are of great interest for experiments with different biological tissues.
\n
\n
\n
\n
\n
4. Analysis of trace element changes in mice treated with CoCl2
\n
Transition-metal cobalt is an essential trace element required for vitamin B12 biosynthesis, enzyme activation, etc., but is toxic in high concentrations. We estimated the effect of cobalt chloride (CoCl2) on relative content of different metal ions in mouse plasma using TJP-AES and on the total protein content [29, 30]. Freeze-dried plasma (2–3 mg) was available for the TJP-AES analysis. On average the relative content of different elements in the plasma of 2-month-old mice balb/c (control group) decreased in the order: Ca > Mg > Si > Fe > Zn > Cu ≥ Al ≥ B. The 60 days treatment of mice with CoCl2 (daily dose 125 mg/kg) did not change appreciably the relative content (ReCo) Ca, Cu and Zn, while a 2.3-fold significant decrease in the ReCo of B and a significant increase in the content of Si (3.4-fold), Fe and Al (2.1-fold) and Mg (1.5-fold) was found (Table 6). The ReCo of Mo and Co for untreated mice was lower than test sensitivity. Mo in a detectable amount was determined only for two mice in the control group, but the plasmas of 9 out of 16 mice of analyzed group contained this metal. Cobalt treatment resulted in a 2.2-fold decrease in the concentration of total plasma protein and in 1.7-fold immunoglobulins. Clarification of the complex effects of Co2+ on its interactions in vivo with other trace elements is important for the explanation of cobalt toxicity and disturbances in homeostasis and physiological processes such as development, growth, weight gain, immunity, reproduction, etc. [29].
Relative content of different chemical elements and total protein in the freeze-dried blood plasma of control mice and animals treated with CoCl2 [29].
For each mouse, the mean of three repeats is used.
\n
Homogeneous IgGs purified from sera of mice treated (t-IgGs) and non-treated (nt-IgGs) with CoCl2 containing intrinsically bound metal ions hydrolyze DNA with very low activity and lose this activity in the presence of EDTA [30]. The average relative DNase activity (RAs) of nt-IgGs increased after addition of external metal ions in the following order: Zn2+ < Ca2+ < Cu2+ < Fe2+ < Mn2+ < Mg2+ < Co2+ < Ni2+. Interestingly, t-IgGs showed lower activity than nt-IgGs in the absence of external metal ions (2.7-fold) as well as in the presence of Cu2+ (9.5-fold), Co2+ (5.6-fold), Zn2+ (5.1-fold), Mg2+ (4.1-fold), Ca2+ (3.0-fold) and Fe2+ (1.3-fold). But t-IgGs were more active than nt-IgGs in the presence of Ni2+ (1.4-fold) and especially Mn2+ (2.2-fold), which are the best activators of t-IgGs. These data may be useful for an understanding of Co2+ toxicity, its effect on a change of metal-dependent specificity of mouse abzymes [30].
\n
\n
\n
5. Abzymes with oxidation-reduction activities
\n
First, we have estimated the content of metals in the lyophilized plasmas of healthy Wistar rats (Table 7) by the TJP-AES method [31]. The relative amount of metals in the rat plasma decreased in the order: Ca > Mg > Fe > Cu ≥ Zn > Al ≥ Sr. > Ti ≥ Mo ≥ Mn (Table 7). Nine plasmas of healthy Wistar rat’s sera were used for purification of electrophoretically and immunologically homogeneous IgGs according to [31–35]. Homogeneous IgGs according to data of the TJP-AES method did not contain a detectable amount of Sr and Mo (Table 8) [31]. The relative amount of different metals bound to IgGs of SLE and MS patients in average decreased in the following order: Ca ≥ Zn ≥ Ti ≥ Mg ≥ Al ≥ Fe ≥ Cu ≥ Ni > Mn (Table 8). Thus, IgGs of individual rats can interact with metal ions showing a significant difference (Table 8) in spite of their comparable concentrations in the plasmas (Table 7).
The relative content of different trace elements and metals in the lyophilized blood plasmas from nine rats.
The content was determined by TJP-AES method; the relative standard deviation of the results from two experiments were within 5–7%.
The maximal and minimal values for each metal are marked in bold.
Mean ± S.D.
Sign ≤ in all cases means that the presence of metal in the samples is reliable, but its exact concentration cannot be determined; it can be in the range 0.1–1 μg/g.
\n
\n
\n
\n
\n\n
\n
Metal
\n
Relative content (μg/g)
\n
\n
\n
sle-IgGmix
\n
ms-IgGmix
\n
\n\n\n
\n
Al
\n
7.0
\n
8.0
\n
\n
\n
Ca
\n
10
\n
120
\n
\n
\n
Cu
\n
8.0
\n
4.0
\n
\n
\n
Fe
\n
4.0
\n
9.0
\n
\n
\n
Mg
\n
4.0
\n
17.0
\n
\n
\n
Mn
\n
0.2
\n
~0
\n
\n
\n
Ni
\n
6.5
\n
0.7
\n
\n
\n
Ti
\n
2.0
\n
27.0
\n
\n
\n
Zn
\n
37.0
\n
11.0
\n
\n\n
Table 8.
The relative content of metal ions in the lyophilized sle-IgGmix and MS-IgGmix samples from the sera of patients with SLE and MS, respectively*.
Preparations sle-IgGmix and ms-IgGmix are mixtures of equal amounts of electrophoretically homogeneous IgGs from the sera of 12 SLE (sle-IgGmix) and 12 MS (ms-IgGmix) patients.
The content was determined by TJP-AES method; the errors of the values from two experiments were within 5–7%.
\n
We have shown that rat IgGs lose most bound metal ions during the purification [31, 33]. From 26 to 39% of rat IgGs can interact with less or more efficiently with different metals ions. Interestingly, chromatography of IgGs from human plasmas on Chelex non-charged with metal ions led to the binding of a small amount of IgGs (~ 5%) bound with metal ions [36]. Chelex charged with Cu2+ ions additionally adsorbed ~ 38% of the total IgGs. In a number of publications, it was shown that all many catalytic activities are the intrinsic properties of mammalian antibodies and are not caused by impurities of any canonical enzymes [37–46]. For this purpose, in all cases, we have checked several previously developed strict criteria proving that all activities of Abs from blood sera and healthy donors and autoimmune patients belong to the Abs [37–46].
\n
All higher organisms generate energy due to aerobic respiration, the process including a four-electron stepwise reduction to water of molecular oxygen [47–51]. The partially reduced species include OH•, H2O2 and O2•−, are typical oxidants attacking proteins, lipids, DNA and other components of different cells. Oxidative damage of cells components was regarded as the significant factor of carcinogenesis and aging [47, 49, 51].
\n
Antioxidant enzymes (catalases, superoxide dismutases and glutathione peroxidases) are very important for preventing oxidative stress [52–56]. However, these enzymes are located inside of cells, and they undergo rapid inactivation in the blood [54]. Immunoglobulins (Igs) are significantly more stable molecules of blood. Therefore, it was interesting how metal ions can activate oxidation-reduction reactions catalyzed by antibodies. The catalysis of such reactions by the majority of canonical enzymes is dependent on metal ions with variable valence [50, 52–55]. First, we have shown that IgGs of healthy Wistar rats oxidize 3,3‘-diaminobenzidine through a peroxidase activity in the presence of H2O2 and due to an oxidoreductase activity in the absence of H2O2 [31–35]. In the external metal ions absence, the specific peroxidase activity of IgGs of rats varied in the range 1.6–26% comparing with horseradish peroxidase (HRP, taken for 100%). The dialysis of IgGs against EDTA completely lost these activities. External metal ions activated significantly both activities of non-dialyzed (ND) and dialyzed (D) IgGs. The relative activities (RAs) in the presence of external Fe2+ or Cu2+ ions were increased up to 13–198% compared with that for HRP [31]. Cu2+ ions alone stimulated significantly both the oxidoreductase and peroxidase activities of dialyzed D-IgGs, but only at high concentration (≥2 mM) [31]. Mn2+ ions were weakly activated peroxidase activity but at >3 mM Mn2+ was good cofactor of the oxidoreductase activity at a low concentration (<1 mM). Fe2+-dependent peroxidase activity of D-IgGs was revealed at 0.1–5 mM, but Fe2+ cannot activate their oxidoreductase activity. Al3+, Mg2+, Zn2+, Ca2+, and especially Ni2+ and Co2+ were not able to activate D-IgGs, but slightly activated ND-IgGs containing different intrinsic metal ions. Some metal ions activated IgGs especially ND-Abs in accordance with biphasic curves, which were specific for every individual Ab preparation [31]. The combinations of Fe2+ + Zn2+, Fe2+ + Mn2+, Cu2+ + Mn2+ and Cu2+ + Zn2+ and other metal ions led to the oxidation of substrates mainly with single-phase curves. In parallel to a significant increase of the activities comparing with Fe2+, Cu2+ or Mn2+ taken separately, the RAs of the oxidation reactions catalyzing by non-dialyzed and dialyzed IgGs, became to be comparable. Ni2+, Mg2+ and Co2+ sufficiently activated the Cu2+-dependent oxidation of substrates catalyzed by D-IgGs, while Ca2+ inhibited these reactions [31].
\n
As mentioned above, the dependencies of the oxidoreductase and peroxidase activities of the ND-IgGs and D-IgGs on the concentrations of Fe2+, Cu2+ and Mn2+ were biphasic. This indicates that two different metal ions are likely to participate in the catalysis of these reactions. The canonical Cu, Zn superoxide dismutases usually use a Cu2+ ion together with a Zn2+ ion [53–55]. However, the only Cu2+ with variable valency participates in the oxidation of substrates, while Zn2+ serves as a second electrophilic metal cofactor of this enzyme [53–55]. The biphasic dependences can show for a similar function of the same second or another second metal ion. Since D-IgGs and ND-IgGs demonstrate a significantly higher activity in the presence of Cu2+ ions together with Mn2+ or Zn2+ ions, some fractions of IgGs can be Cu/Cu, Cu/Mn or Cu/Zn peroxidases or oxidoreductases. A remarkable increase in the IgGs activity by Cu2+ ions together with Co2+, Mg2+ or Ni2+ can speak in favor that these metal ions can also increase the oxidative function of Cu2+ to some extent as the second ions [31]. Only Fe2+ taken separately was activated the peroxidase activity of D-IgGs at low concentrations (<1 mM). However, FeCl2 was completely unable to activate the oxidoreductase activity of D-IgGs. Most probably, Cu2+ + Mn2+ is an optimal pair for both the peroxidase and oxidoreductase reactions. It seems more likely that the activation of IgGs by two metal ions with variable oxidation state proceeds either using the second metal as an electrophilic cofactor.
\n
It was demonstrated that small fractions of IgGs from the sera of healthy humans as well as their Fab and F(ab)2 fragments oxidize DAB through peroxidase and oxidoreductase activities [36]. In contrast to rat antibodies, IgGs from human blood have both the dependent and independent on metal ions activities. After dialysis of human IgGs against EDTA and EGTA, the relative peroxidase and oxidoreductase activity dependently of IgG preparation decreased from 100 to ~10–85 and 14–83%, respectively. Addition of external metal ions to D-IgGs and ND-IgGs results in a significant increase in their activities. Separation of IgGs on Chelex results in Abs separation to many different subfractions with different affinities to the chelating resin. In the presence of Cu2+ external ions, the specific peroxidase RA of several human IgG subfractions after chromatography achieves 20–27% comparing with horseradish peroxidase (HRP, taken for 100%). The oxidoreductase activity of many IgG subfractions is ~ 4–6-fold higher than that for HRP [36].
\n
It was shown, that IgGs of rats and humans effectively oxidize not only DAB but also many other toxic, carcinogenic and mutagenic compounds such as phenol, o-phenylenediamine, α-naphthol, p-hydroquinone, etc. [34]. However, overall, the relative peroxidase and oxidoreductase activities of polyclonal rat IgGs in the presence of different metal ions is ~ 10–100-fold higher than those of polyclonal human IgGs. Interestingly, rats are known as the most resistant mammals to all harmful factors of an environment including carcinogens, mutagens and radiation. One cannot exclude that this is due to better protection of rats compared to peoples from harmful factors due to more active metal-dependent Abs with peroxidase and oxidoreductase activities.
\n
\n
\n
6. Dependence of DNA-hydrolyzing abzymes on metal ions
\n
It was shown in many articles that electrophoretically and immunologically homogeneous polyclonal IgGs from sera of healthy volunteers and experimental mice are not active in the hydrolysis of DNA and RNA (for review see [37–46]). The occurrence of auto-Abs with catalytic activities is a distinctive feature of mammalian autoimmune diseases (reviewed in [37–46]). IgGs and/or IgMs abzymes hydrolyzing DNA and RNA were revealed in the sera of patients with several autoimmune and viral pathologies: SLE [57–61], multiple sclerosis [62–64], Hashimoto’s thyroiditis and polyarthritis [65, 66], schizophrenia [67] and with three viral diseases viral hepatitis [68], acquired immunodeficiency syndrome [69] and tick-borne encephalitis [70]), as well as human milk [71–73], SLE mice [74, 75] and experimental autoimmune encephalomyelitis(EAE) mice [76, 77]. Antibodies with DNase activity from the blood of patients and mice with various diseases were dependent on different metal ions [57–70, 74–77], while human milk contains metal-dependent and independent DNase sIgAs and IgGs [71–73]. The RAs of IgGs from the sera of patients (and mice) with different AIDs vary significantly from patient to patient [57–70, 74–77]. Figure 6 shows the cleavage of plasmid supercoiled (sc)DNA by 10 various IgGs bound with internal metal ions from the sera of patients with various autoimmune diseases.
\n
Figure 6.
Relative DNase activities of catalytic IgG-abzymes from sera of 10 different patients with various diseases in the hydrolysis of scDNA. Lanes 1–10 correspond to IgGs of 10 different patients; C1, DNA incubated alone; C2 and C3, DNA incubated with Abs of two healthy donors.
\n
During this time, some IgGs cause only single breaks in one strand of scDNA converting it to the relaxed form (lanes 1–3), when others make multiple breaks forming DNA linearization (lanes 4–6). The most active IgGs hydrolyze scDNA into medium- and short-length oligonucleotides (lanes 7–10). Polyclonal DNase IgGs from the sera of autoimmune-prone MRL mice were not after Abs dialysis against EDTA, but were activated by different externally added metal (Me2+) ions: Mn2+ ≥ Mg2+ > Ca2+ ≥ Cu2+ > Co2+ ≥ Ni2+ ≥ Zn2+ [74, 75]. Fe2+ ions could not stimulate the hydrolysis of scDNA by the Abs. The initial rate dependencies on the concentration of different Me2+ ions were mostly bell-shaped, having from one to four maxima at different concentrations of Me2+ ions. Mn2+, Ni2+ and Co2+ activated DNA hydrolysis. The Mn2+-dependent scDNA hydrolysis was activated by Ni2+, Ca2+, Co2+ and Mg2+, but was inhibited by Zn2+ and Cu2+. Only in the case of Mg2+and Mg2+ or Ca2+ as the second metal ions, an accumulation of linear DNA was observed. Affinity chromatography on DNA-cellulose separated DNase mouse IgGs to many subfractions having different affinities for DNA and varying levels of the relative activity (0–100%) in the presence of Mn2+, Ca2+ and Mg2+ ions. In contrast to all human DNases having 1 pH optimum, mouse IgGs hydrolyzing DNA showed several pronounced pH optima from 4.5 and 9.5; in the presence of Ca2+, Mn2+ and Mg2+ ions, these dependencies were different. These findings show the extreme diversity of the ability of metal-dependent mouse IgGs functioning at different pHs and to be activated by various optimal metal cofactors. At the same time, a similar situation on an extreme diversity of Me2+-dependent Abs was observed for DNase abzymes from sera of the patient with different autoimmune and viral diseases [70] including monoclonal light chains of human IgGs [78] (e.g., Figure 7).
\n
Figure 7.
The RAs of MBP-hydrolyzing activity of twenty-two MLChs after their treatment with specific inhibitors of various type proteases. Different MLChs were preincubated in the absence of inhibitors (black bars, control-C), with 50 mM EDTA (gray bars) or 1.0 mM PMSF (white bars) before addition to the standard reaction mixture (A and B). Panel C demonstrates several examples of the RAs of MLChs with metal-dependent (1, 5, 12, 15, and 21) and serine-like activity (4 and 11), which no changing their activity after treatment with iodoacetamide; three MLChs (10, 14, and 18) showing negative response to EDTA and PMSF as well as MLCh-22 having positive answer to PMSF and EDTA after their preincubation with iodoacetamide resulting a significant decrease in the protease activity. Gray and white bars (panel C) correspond respectively to the activity after and before (control) these preparation treatment with iodoacetamide. The Ras of all MLChs before their treatment with different specific inhibitors was taken for 100 %.
\n
Dependently on patient demonstrated different substrate specificity [63]. All the data obtained showed that polyclonal MS IgGs could contain different combinations of sequence-independent and sequence-dependent endo- and exonuclease activities [63]. The enzymatic properties of the DNA- and RNA-hydrolyzing IgGs of patients with various AIDs [37–46] distinguished them from all known canonical DNases and RNases [79–81].
\n
Polyclonal DNase IgGs from sera of autoimmune patients, SLE mice, rabbits immunized with DNA and human milk are usually very heterogeneous in their affinity for DNA and can be separated into many subfractions by chromatography on DNA-cellulose [78, 82, 83]. An immunoglobulin light chain phagemid library was prepared using peripheral blood lymphocytes of patients with SLE [78, 82, 83]. Phage particles displaying light chains interacting with DNA were isolated by chromatography on DNA-cellulose; the fraction eluted by 0.5 M NaCl and acidic buffer (pH 2.6) were used for obtaining of individual monoclonal light chains (MLChs, 27–28 kDa) [78, 82, 83]. About 45 of 451 and 33 of 687 individual colonies corresponding to peaks eluted with 0.5 M NaCl and acidic buffer, respectively, were randomly chosen for a study of MLChs with DNase activity. About 15 of 45 (Km = 260–320 nM) and 19 of 33 (Km = 3–9 nM) MLChs in the first and second case efficiently hydrolyzed DNA. All 34 MLChs demonstrated different optimal concentrations of KCl or NaCl and pH optima. All MLChs were metal-dependent DNases. The ratio of relative DNA-hydrolyzing activity in the presence of different metal ions was individual for each MLCh. For example, for monoclonal kappa light chain NGK-1 in optimal conditions the RAs decreased in the following order (%): Mn2+ (26.3) ≥ Ca2+ (23.0) ≥ Mg2+ (21.0) > Ni2+ (15.0) > Zn2+ (11.4) > Cu2+ (2.9) > Co2+ (0.0) [83]. But in average, the activity in DNA hydrolysis for all MLChs decreased in the following order: Mn2+ > Co2+ > Mg2+ > Ni2+ ≈ Ca2+ > Cu2+ > Zn2+ [78, 82, 83].
\n
It is known, that Co2+, Mn2+, Ca2+ and Ni2+ activate mammalian DNase I in much lesser degree than Mg2+ ions [80, 81]. Interestingly, human milk polyclonal sIgA DNase abzymes mainly Me2+-independent and they were only slightly activated by Mg2+, Mn2+ or Zn2+, and the cleavage of DNA substrates was inhibited by Ca2+ and Cu2+ [73]. The effect of metal ions on DNase activity of intact Abs from sera of MS patients decreased in the order: Mn2+ > Mg2+ > Zn2+ > Ca2+ [84]. The DNA-hydrolyzing activity of tick-borne encephalitis IgGs decreased in the following order: Mn2+ ≥ Co2+ ≥ Mg2+ > Ca2+, while Zn2+, Ni2+ and Cu2+ did not stimulate DNA hydrolysis [70]. Polyclonal intact IgGs from MRL mice following specific order of DNase activity activation by different metal ions: Mn2+ ≥ Mg2+ > Ca2+ ≥ Cu2+ > Co2+ ≥ Ni2+ ≥ Zn2+ [74]. Thus, the relative activity of metal-dependent abzymes hydrolyzing DNA depends on the type and timing of the disease as well as can be specific for every individual patient (or animal). On overall, relative metal-dependent DNase activity in blood of patients with different autoimmune and viral diseases increases in the following order: Diabetes < Viral hepatitis ≈ Tick-borne encephalitis < Polyarthritis ≤ Hashimoto’s thyroiditis < Schizophrenia < AIDS ≤ Multiple sclerosis < SLE [85].
\n
\n
\n
7. Dependence of RNA-hydrolyzing abzymes on metal ions
\n
First, it was shown that mouse SLE monoclonal IgGs directed against different DNA could effectively hydrolyze both DNA and RNA and cleavage of RNAs is 30–100-fold faster than DNA [86]. Later we have shown that immunization of rabbits with DNA, RNA, RNase A, DNase I or DNase II leads to the formation of abzymes that hydrolyze both DNA and RNA [85]. Interestingly, the substrate specificities of RNase IgGs from patients with autoimmune thyroiditis and polyarthritis [65], SLE [58], MS [62] and hepatitis [68] for different classic homopolynucleotides, cCMP and tRNAPhe with a stable compact structure [39, 59] were different and correlated with the type of disease and were well distinguishable from those of canonical RNases. The activity was strongly dependent on the patient and its disease, but in average increased in the order: hepatitis < polyarthritis < autoimmune thyroiditis < SLE ≤ MS. Abzymes of patients of SLE and MS patients demonstrate new RNase activity stimulated by Mg2+ ions [39, 59, 65, 87]. In the presence of Mg2 + ions, the abzymes produced products corresponding to new cleavage sites of mutant tRNALys, indicating its local structural or conformational changes compared to tRNALys from mitochondria. Thus, different metal ions play a very important role in the functioning of abzymes with DNase and RNase activities.
\n
\n
\n
8. Dependence of protein-hydrolyzing abzymes on metal ions
\n
For the first time, elevated levels of polyclonal antibodies to myelin basic protein (MBP) and abzymes hydrolyzing MBP were detected in the blood of MS [88–91] and then of SLE [92–95] patients. In the blood of healthy donors, no such abzymes have been detected [88–95]. It is believed that the mechanism of the pathogenesis of MS is associated with the destruction of myelin (including MBP), leading to inflammation processes associated with autoimmune reactions [96]. Some immunological and biochemical indicators of patients with MS and SLE are very similar [45]. First, we have shown that polyclonal IgGmix (a mixture of equimolar IgGs from 10 MS patients) can hydrolyze MBP in the presence better, than in the absence of different metal ions [88–91]. According to TJP-AES data, homogeneous IgG preparations of MS patients contained several intrinsic metal ions; Fe ≥ Ca > Cu ≥ Zn ≥ Mg ≥ Mn ≥ Pb ≥ Co ≥ Ni [90]. Then, a minor Me2+-dependent fraction was obtained by chromatography of one IgG preparation on Chelex-100. This IgG fraction could not hydrolyze MBP in the absence of metal ions but was activated after addition of external Mg2+ > Mn2+ > Cu2+ > Ca2+ [90]. Proteolytic activities of individual IgGs from other MS patients were also activated by Fe2+, Ni2+, Zn2+, Co2+ and Pb2+, and especially Ni2+. Interestingly, specific proteolytic metal-dependent and independent activities of IgMs and IgAs from sera of MS patient were usually higher than those of IgGs [89]. A significant diversity of different fractions of polyclonal MS IgGs in their affinity for MBP and the hydrolysis of MBP at different optimal pHs (3–10.5) was demonstrating [91]. IgGs containing kappa- and lambda-light chains showed comparable RAs in the hydrolysis of MBP. IgGs of all four sub-classes were active, with their different average contribution to the total activity of abzymes in the hydrolysis of MBP: IgG1 (1.5–2.1%) < IgG2 (4.9–12.8%) < IgG3 (14.7–25.0%) < IgG4 (71–78%) [91]. The properties of MS abzymes demonstrating their significant catalytic diversity distinguish them from all known mammalian proteases including metal-dependent ones. These abzymes can attack MBP of the myelin-proteolipid shell of axons and play an important role in MS pathogenesis [88–91].
\n
At the initial stage of SLE development antibodies against DNA, as well as DNA- and RNA-hydrolyzing antibodies are mostly developed [37–46]. A little later, however, similar to MS pathology in the case of SLE patients the production of Abs against MBP and abzymes hydrolyzing this protein is happening [92–95]. The relative content of different metal ions in the preparations of lyophilized sle-IgGmix and ms-IgGmix from sera of patients with MS and SLE estimated by TJP-AES method to some extent comparable (Table 8). Ca2+ was the best activator of SLE-IgGmix and its activity increased in the order: Ca2+ > Mg2+ ≥ Co2+ ≥ Fe2+ ≥ Ni2+ ≥ Cu2+ ≥ Mn2+. Zn2+ inhibits the activity, while Fe2+ cannot activate sle-IgGmix. Ms-IgGmix before dialysis against EDTA showed another order of the activity: Mg2+ > Mn2+ ≥ Cu2+ ≥ Ni2+ ≥ Co2+ ≥ Ca2+, while Fe2+ and Zn2+ slightly inhibit its activity. Thus, on average, patients with MS and SLE develop abzymes hydrolyzing MBP with different dependence on various metal ions. Combinations of Ca2+ + Co2 and Ca2+ + Mg2+ results in a significant increase in the MBP-hydrolyzing activity comparing to Ca2+, Mg2+ and Co2+ or ions taken separately [92]. Lambda-IgGs demonstrated higher RAs in the hydrolysis of MBP than kappa-IgGs [93]. The pH profiles of IgG4, IgG3, IgG2, IgG1 of SLE patients were unique; their RAs increased in the order: IgG4 < IgG2 < IgG3 < IgG1. Thus, the immune systems of SLE similarly to MS patients produce a variety of metal-dependent anti-MBP abzymes, which can hydrolyze MBP of the myelin-proteolipid shell of axons and can play important role in the pathogenesis of MS and SLE patients [45, 46].
\n
Phagemid library derived from lymphocytes of peripheral blood of patients with SLE was used for obtaining of MLChs hydrolyzing MBP [97–100]. About 22 of 72 MLChs hydrolyzing only MBP (not other control proteins) having various pH optima in a 5.7–9.0 range and different specificity in the hydrolysis of four various MBP oligopeptides [97]. Eleven MLChs were metalloproteases, while four and three MLChs showed serine-like and thiol-like proteolytic activities, respectively. The activity of three MLChs was suppressed by both PMSF and EDTA, while the other two by EDTA and iodoacetamide and one by EDTA, PMSF and iodoacetamide. The ratio of RAs in the presence of Mg2+, Ca2+, Mn2+, Zn2+, Cu2+, Ni2+ and Co2+ was very specific for all metal-dependent MLChs. For the total preparation of MLChs, the activity decreased in the order: Са2+ ≥ Сo2+ ≈ Мg2++ ≥ Mn2+ ≥ Ni2 ≈ Cu2+ ≈ Zn2+ [97].
\n
In addition to these 22, were isolated other 3 MLChs, which were analyzed in more detailed. NGTA1-Me-pro (MLCh-23) was a typical metalloprotease inhibited only by EDTA [98]. The activity of MLCh-23 in the hydrolysis of MBP was reduced in the presence of ions of seven different metals in the following order: Са2+ > Мg2+ > Ni2+ ≥ Zn2+ ≥ Сo2+ ≥ Mn2+ > Cu2+. MLCh-23 has two active sites into the light chain with very distinct pH optima: pH 6.0 and 8.5 and different affinity for MBP [98]. Specific inhibitors of NGTA2-Me-pro-Tr (MLCh-24) were PMSF (42%) and EDTA (58%): it exhibits properties of a chimeric protease with serine and metal-dependent activities [99]. The addition of ions of different metals led to a decrease in the activity of MLCh-24 in the following order: Са2+ ≥ Mn2+ ≥ Мg2+ ≈ Сo2+ ≈ Ni2+ ≥ Cu2+ ≥ Zn2. NGTA2-Me-pro-Tr is the first example of an MLCh-23 having two combined centers with serine and metalloprotease activities.
\n
It should be noted that all recombinant MLChs were obtained by affinity chromatography of phage particles on MBP-Sepharose. Taking this into account, a very unexpected result was obtained from analysis of NGTA3-pro-DNase (MLCh-25) [100]. Only 1 MLCh-25 of 25 recombinant MLChs effectively hydrolyzed not only MBP but also DNA. Preincubation of MLCh-25 with both PMSF (67%) and EDTA (36%) resulted in suppression of its protease activity. Ions of different metals activated MLCh-25 in the following order: Са2+ ≥ Ni2+ > Сo2+ ≈ Mn2+ ≥ Cu2+ ≈ Zn2+ ≥ Mg2+ [100]. The affinity of MLCh-25 metal-dependent and serine-like active centers for BMP was different. The DNase activity of MLCh-25 decreases in the following order: Mn2+ ≈ Сo2+ ≥ Мn2+ > Сu2+ ≈ Ni2+ ≥ Са2+ > Zn2+, which completely distinguishes MLCh-25 from canonical DNases [72]. Metal-dependent casein hydrolyzing sIgA antibodies from human milk were described [101]. The RA of sIgAs after removal of intrinsic metal ions increase their activity in the presence of external Fe2+ > Ca2+ > Co2+ ≥ Ni2+ and especially combinations of metals: Co2++Ca2+ < Mg2+ + Ca2+ < Ca2+ + Zn2+ < Fe2+ + Zn2+ < Fe2+ + Co2+ < Fe2+ + Ca2+ [101].
\n
\n
\n
9. Catalytic activities of antibodies of HIV-infected patients
\n
Metal-dependent IgGs and/or IgMs from the blood of HIV-infected patients hydrolyzing DNA [69], viral reverse transcriptase [102] and integrase [103–105], and all histones [106] were described. Average activities of anti-IN IgGs in the hydrolysis of IN decreased in the order Mn2+ > Mg2+ ≈ Cu2+ > Co2+ while for IgMs in another order Cu2+ > Mn2+ > Co2+ ≫ Mg2+. Our findings show that active centers of anti-IN polyclonal abzymes of AIDS patients can contain amino acid residues providing thiol, serine, acidic and metal-dependent proteases. But the ratio of these abzymes activities may be individual for every HIV-infected patient.
\n
In addition, IgGs from sera of HIV-infected patients hydrolyze all human histones [106]. The RAs of IgGs in the hydrolysis of histones (H4, H3, H2a, H2b and H1) varied significantly for Abs of different patients. The effects of different external metal ions on the dialyzed polyclonal IgGs in the hydrolysis of five individual histones were very different. For example, maximal activation of one IgG preparation was observed in the hydrolysis of H4 by Zn2+ and Ni2+, H3 by Cu2+ and Ni2+, H2a by Cu2+, H2b by Co2+ and Ni2+, H1 by Cu2+ and Mn2+. Such an exceptional diversity in activation by different metals ions was observed for all 32 IgGs [106]. Importantly, mammalian immune system theoretically can produce up to 106 variants of Abs against one antigenic determinant and all of these Abs may be different.
\n
\n
\n
10. Conclusion
\n
Using the TJP-AES method, we have estimated the relative contents of various trace elements, including metals in various organs, tissues and biological fluids of humans and animals, as well as in immunoglobulins from these sources., the maximal RAs of abzymes with different catalytic activities are most often achieved not in the presence of metal ions, which are contained in biological sources and antibodies in maximum quantities. Some specific abzymes show maximum activity in the presence of metal ions, which are minor elements of different organs and biological fluids. The question is why there are so many abzymes with very different properties including metal-dependent ones against the same protein. First, mammalian immune system theoretically can produce up to 106 variants of Abs against one antigenic determinant and all of these Abs may be different. In addition, proteins and nucleic acids can adsorb ions of various metals including traces elements on their surfaces. Therefore, some specific antibodies (and abzymes) can be against fragments (antigenic determinants) of DNA and proteins containing no metal ions. Some other specific metal-dependent abzymes with nuclease and protease activities can be antibodies against sequences associated with one or more metal ions. In addition, not only antibodies against substrates imitating transition states of chemical reactions can possess catalytic activities, but also anti-idiotypic Abs against active centers of various enzymes. The activity of many various enzymes depends on the ions of different metals. Since secondary—anti-idiotypic antibodies against such active sites should contain all the structural components of an enzyme active center including amino acid residues for binding metal ions, they can be metal-dependent abzymes. In this chapter, we have analyzed not only the relative content of different metal ions in various biological substances but also analyzed a possible function of metal ions in the catalysis by autoantibodies of different chemical reactions.
\n
\n
Acknowledgments
\n
This research was possible due to grant from the Russian Science Foundation (No. 16-15-10,103) to G.A. Nevinsky).
\n
\n',keywords:"two-jet plasma atomic emission spectrometry analysis, minor and trace elements, functional role of metals",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/61016.pdf",chapterXML:"https://mts.intechopen.com/source/xml/61016.xml",downloadPdfUrl:"/chapter/pdf-download/61016",previewPdfUrl:"/chapter/pdf-preview/61016",totalDownloads:1153,totalViews:157,totalCrossrefCites:0,totalDimensionsCites:0,totalAltmetricsMentions:0,impactScore:0,impactScorePercentile:10,impactScoreQuartile:1,hasAltmetrics:0,dateSubmitted:"November 30th 2017",dateReviewed:"February 25th 2018",datePrePublished:null,datePublished:"September 5th 2018",dateFinished:"April 25th 2018",readingETA:"0",abstract:"Microelements play different important roles in many physiological processes in all biological systems in both normal physiological and pathological conditions. They take part in the transport of nutrients and gases, support temperature, acid-base balance, homeostasis of the human organisms, maternal and child mental health, the functioning of enzymes, protein and DNA syntheses, cytoskeleton activation, etc. We have performed simultaneous determination of a number of minor and trace elements in whole blood and tissues of mammals by two-jet plasma atomic emission spectrometry (TJP-AES). TJP-AES allows direct analysis of powders without wet acid digestion and can be used for analysis of both large and small amount of the sample, which is important for biomedical investigations with humans and experimental animals. In addition, a content of different elements in preparations of human immunoglobulins was estimated by TJP-AES as well as using different physicochemical methods, the functional role of metal ions in antibodies functioning was analyzed. The analysis of the relative activity of antibodies with catalytic activity (abzymes) in the hydrolysis of DNA, RNA, proteins, peptides and oxidation-reduction reactions and the role of metal ions in the catalysis of these reactions by abzymes were carried out.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/61016",risUrl:"/chapter/ris/61016",book:{id:"6728",slug:"trace-elements-human-health-and-environment"},signatures:"Natalia P. Zaksas and Georgy A. Nevinsky",authors:[{id:"47119",title:"Dr.",name:"Georgy",middleName:null,surname:"Nevinsky",fullName:"Georgy Nevinsky",slug:"georgy-nevinsky",email:"nevinsky@niboch.nsc.ru",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"52213",title:"Dr.",name:"Natalia",middleName:null,surname:"Zaksas",fullName:"Natalia Zaksas",slug:"natalia-zaksas",email:"zak@niic.nsc.ru",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Two-jet plasma atomic emission spectrometry",level:"1"},{id:"sec_2_2",title:"2.1. A two-jet plasmatron",level:"2"},{id:"sec_3_2",title:"2.2. Analytical regions",level:"2"},{id:"sec_4_2",title:"2.3. Excitation mechanisms",level:"2"},{id:"sec_6",title:"3. Application of TJP-AES",level:"1"},{id:"sec_6_2",title:"3.1. Analysis of high purity substances",level:"2"},{id:"sec_7_2",title:"3.2. Analysis of biological samples",level:"2"},{id:"sec_7_3",title:"Table 2.",level:"3"},{id:"sec_8_3",title:"Table 4.",level:"3"},{id:"sec_9_3",title:"Table 5.",level:"3"},{id:"sec_12",title:"4. Analysis of trace element changes in mice treated with CoCl2",level:"1"},{id:"sec_13",title:"5. Abzymes with oxidation-reduction activities",level:"1"},{id:"sec_14",title:"6. Dependence of DNA-hydrolyzing abzymes on metal ions",level:"1"},{id:"sec_15",title:"7. Dependence of RNA-hydrolyzing abzymes on metal ions",level:"1"},{id:"sec_16",title:"8. Dependence of protein-hydrolyzing abzymes on metal ions",level:"1"},{id:"sec_17",title:"9. Catalytic activities of antibodies of HIV-infected patients",level:"1"},{id:"sec_18",title:"10. Conclusion",level:"1"},{id:"sec_19",title:"Acknowledgments",level:"1"}],chapterReferences:[{id:"B1",body:'Taylor A, Day MP, Hill S, Marshall J, Patriarca M, White M. 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Understanding HIV/AIDS Management and Care—Pandemic Approaches the 21st Century. Rijeka, Croatia: InTech; 2011. pp. 151-192\n'},{id:"B45",body:'Nevinsky GA. Autoimmune processes in multiple sclerosis: Production of harmful catalytic antibodies associated with significant changes in the hematopoietic stem cell differentiation and proliferation. In: Conzalez-Quevedo A, editor. Multiple Sclerosis. Rijeka, Croatia: InTech; 2016. pp. 100-147\n'},{id:"B46",body:'Nevinsky GA. Catalytic antibodies in norm and systemic lupus erythematosus. In: Khan WA, editor. Lupus. Rijeka, Croatia: InTech; 2017. pp. 41-101. DOI: 10.5772/67790\n'},{id:"B47",body:'Ames BN. Dietary carcinogens and anticarcinogens. Oxygen radicals and degenerative diseases. Science. 1983;221:1256-1264. PMID: 6351251\n'},{id:"B48",body:'Cutler RG. Antioxidants and aging. The American Journal of Clinical Nutrition. 1991;53:373S-3379. PMID: 1985414\n'},{id:"B49",body:'Beckman KB, Ames BN. The free radical theory of aging matures. Physiological Reviews. 1998;78:547-581. PMID: 9562038\n'},{id:"B50",body:'Feuers RJ, Weindruch R, Hart RW. Caloric restriction, aging, and antioxidant enzymes. Mutation Research. 1993;295:191-200. PMID: 7507557\n'},{id:"B51",body:'Allen RG. Free Radicals in Aging. Boca Raton, FL: CPC Press; 1993. pp. 12-23\n'},{id:"B52",body:'Mates JM, Perez-Gomez C, Nunez de Castro I. Antioxidant enzymes and human diseases. Clinical Biochemistry. 1999;32:595-603. PMID: 10638941\n'},{id:"B53",body:'Mates JM, Sanchez-Jimenez F. Antioxidant enzymes and their implications in pathophysiologic processes. Frontiers in Bioscience. 1999;4:D339-D345. PMID: 10077544\n'},{id:"B54",body:'Zenkov NK, Lankin VZ, Men\'shikova EB. Oxidative Stress. Biochemical and Patho-physiological Aspects. Moscow: MAIK, Nauka/Interperiodica; 2001. pp. 3-343\n'},{id:"B55",body:'Dhaunsi GS, Gulati S, Singh AK, Orak JK, Asayama K, Singh I. Demonstration of Cu-Zn superoxide dismutase in rat liver peroxisomes. 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Systemic lupus erythematosus: Molecular cloning and analysis of recombinant monoclonal kappa light chain NGTA2-Me-pro-Tr possessing two different activities-trypsin-like and metalloprotease. International Immunology. 2015;27:633-645. DOI: 10.1093/intimm/dxv042\n'},{id:"B100",body:'Timofeeva AM, Buneva VN, Nevinsky GA. SLE: Unusual recombinant monoclonal light chain ngta3-pro-dnase possessing three different activities trypsin-like, metalloprotease and DNase. Lupus: Open Access. 2017;2(2):1-12\n'},{id:"B101",body:'Odintsova ES, Zaksas NP, Buneva VN, Nevinsky GA. Metal dependent hydrolysis of β-casein by sIgA antibodies from human milk. Journal of Molecular Recognition. 2011;24:45-59. DOI: 10.1002/jmr.1022\n'},{id:"B102",body:'Odintsova ES, Kharitonova MA, Baranovskii AG, Sizyakina LP, Buneva VN, Nevinsky GA. Proteolytic activity of IgG antibodies from blood of acquired immunodeficiency syndrome patients. Biochemistry (Moscow). 2006;71:251-261. 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Institute of Chemical Biology and Fundamental Medicine, Siberian Division, Russian Academy of Sciences, Russia
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1. Introduction
Studies on heat transfer in porous media have been augmented rapidly. The porous media likely filled with metallic foams with open cells, metallic sintered fiber felts, or metallic lattice frame structures have high thermal conductivity and have much importance in different engineering areas such as heat transport enhancement, thermal storage, solar thermal utilization, modeling of biological tissues, etc. There are two main models, the local thermal equilibrium (LTE) and the local thermal non-equilibrium (LTNE), which can be used to represent heat transfer phenomena in a porous medium. The LTE model can be used effectively to examine the heat transfer in porous media when the temperature difference between the fluid phase and that of the solid phase is small. But not in all situations the differences between temperatures are ignored. In these cases, the influence of the interfacial surface and interstitial convective heat transfer coefficient became major factors of heat exchange among the fluid and solid phases. In such cases, LTNE model needs to be utilized.
A number of researchers investigated heat transfer through LTE model in porous media that are made of low-conductive materials. Poulikakos and Kazmierczak [1] present a fully developed convective flow in a channel with partially filled porous matrix. They modeled the problem of heat and mass flow in the porous medium by using the Brinkman-LTE model to see the effect of parameters on the flow field and on the heat. Vafai and Kim [2] investigated the forced convection flow in a porous medium channel through Forchheimer-LTE model and found a significant increase in the rate of heat transfer as the inertia parameter increases especially for high- to medium-permeability porous media. Gong et al. [3] studied the influence of the thermal conductivity, permeability, and the porous material’s thickness on heat transfer by using a Brinkman-LTE model in annular duct. Their obtained results display that heat transfer is improved by increasing either the thermal conductivity or the permeability. Cheng and Hsu [4] studied the heat and mass flow in an annulus enclosed with porous media by using Brinkman-LTE model under the influence of permeability and porosity. Mitrovic and Maletic [5] got the results for heat and mass flow in a parallel-plate porous channel by using the LTE model with respect to asymmetrical conditions. Sheikholeslami et al. [6] investigated the free convection flow in permeable enclosure by using Darcy-LTE model. In another study, he used the Darcy-LTE model for free convection flow in porous cavity [7].
In highly thermal conductive porous media, solid has usually higher thermal conductivity three to five times more than the fluid. In this case, the LTE model no longer satisfies the requirements of modeling. Then two-energy equation LTNE model is used to measure accurate temperature distribution in fluid and solid phases. Kuznetsov [8] got the results of heat transport phenomena in narrow annulus by using Darcy-LTNE model. Xu et al. [9] studied the Forchheimer-LTE/Forchheimer-LTNE models for heat transfer in pore channel having different thermal conductivities of fluid and solid phases and found the maximum heat distribution in the case of LTE model as compared to LTNE model. Lu et al. [10] described the force convection flow in cylinder fill with pore spume by using Brinkman-LTNE models. Zhao et al. [11] discussed the heat and mass flow through porous media in annulus by using Brinkman-LTNE. Ouyang et al. [12] briefly studied the heat transport phenomena in equidistant-plate channel fill with porous matrix. Xu et al. [13] investigated the mass flow by Brinkman model and LTNE model for heat transfer in equidistant plates filled with mini-spume. Shaikh and Memon [14] provided the numerical results for heat transport in round pipe with or without pore medium by using Darcy-Brinkman-Forchheimer models along LTNE model. Sheikholeslami and Houman [15] reported the transportation of fluid inside a porous cavity through LTNE model.
An important heat transfer fluid is nanofluid that is used in industries because of high rates of heat transfer. The main purpose of nanofluids is to achieve great enhancement in thermal or rheological properties. In a continuation of achieving better thermal conductivity and thermal performance of nano-material, many studies have been done [16, 17, 18, 19, 20]. Recently, Sheikholeslami [21] demonstrated the nanofluid flow in a porous enclosure by Darcy law model. In another study, he investigated the nanofluid flow in a porous media through non-Darcy law model [22]. In current study, keeping in mind these thermal properties of nanofluid, its fully developed convective flow in a parallel-plate channel filled with highly porous media is investigated. For thermal transport in porous media, LTE and LTNE models are utilized with Darcy and Brinkman models. In addition, the temperature profiles under nanoparticle concentration for both different approaches of heat transfer with constant heat flux at wall are calculated and compared.
2. Modeling and formulation
Consider the fully developed forced convection flow of nanofluid through a parallel-plate porous channel filled with open-celled metallic foams. The schematic diagram of the channel is shown in Figure 1.
Figure 1.
Geometry of problem.
Two infinite plates with height 2H are exposed by constant heat flux qw. In this problem, Darcy-LTE, Brinkman-LTE, Darcy-LTNE, and Brinkman-LTNE models are employed for mass flow and heat transfer process in porous media with hydraulically and thermally fully developed conditions.
For fully developed fluid flow, momentum equation with Brinkman term is
0=−dpdx+μeεd2udy2−μnfKu.E1
Here, u is the velocity, μe is the effective viscosity, μnf is the viscosity of nanofluid, K is the permeability, and ε is the porosity.
Temperature distribution in porous media owns two basic models LTE and LTNE. The LTE model containing one energy equation that treats the same value of temperature for fluid and solid phases is given in Eq. (2), while the LTNE model having two-energy equations that treats the different values of the temperatures for solid and fluid phases is shown in Eqs. (3) and (4):
ρCpeu∂Tf∂x=ke∂2Tf∂y2,E2
0=kse∂2Ts∂y2−hATs−Tf,E3
ρCpeu∂Tf∂x=kfe∂2Tf∂y2+hATs−Tf.E4
In the above equation, Tf is the temperature of fluid phase, Ts is the temperature of solid phase, h is the heat transfer coefficient, A is the specific surface area, ρCp is the heat capacity, ke is the effective thermal conductivity, kfe is the fluid phase’s thermal conductivity, and kse is the solid phase’s thermal conductivity.
For corresponding boundary conditions, the temperature of the solid and the fluid at the wall interface will be the same, and velocity is considered to be zero:
y=H:u=0,Tf=Ts=Tw,y=−H:u=0,Tf=Ts=Tw,E5
where Tw is the temperature at interface.
The total heat flux qw is shared among the solid and fluid phases’ subject to their temperature gradients and effective conductivities at the wall:
In this case, Darcy model for fluid flow and LTNE model for temperature distribution in solid and fluid phases are utilized. In the Darcy model, the velocity distribution is taken to be uniformed. So, the energy equations for solid and fluid phases are dimensionless as
kseksd2θsdY2−Dθs−θf=0,E9
Ckfekfd2θfdY2+Dθs−θf=1.E10
2.2 Darcy-LTE model
In this case, the energy equation for the LTE model is normalized as
Ckekfd2θfdY2=1E11
2.3 Brinkman-LTNE model
In this part, the Brinkman and LTNE models for heat and mass transfer are utilized. The dimensionless governing equations are obtained as
1εμeμfd2UdY2−s2μnfμfU+P=1,E12
kseksd2θsdY2−Dθs−θf=0,E13
Ckfekfd2θfdY2+Dθs−θf=U.E14
2.4 Brinkman-LTE model
For the Brinkman and LTE models, governing equations in dimensionless form are
In the above equations, the effective viscosity μe is defined as
μe=1+2.5εμnf,E18
where
μnf=μf1−ϕ2.5.E19
In the above equation, ϕ is the nanoparticle volume fraction and μf is the viscosity of base fluid.
Since the heat is transferred via nanofluid in porous media, the effective thermal conductivity is as follows:
ke=kmknfεkm+1−εknf,E20
kfe=εknf,E21
kse=1−εknf.E22
In the above equation, knf is given as
knf=kp+2kf+2kp−kfϕkp+2kf−kp−kfϕkf,E23
where kf is the thermal conductivity of fluid, kp is the thermal conductivity of nanoparticle, and km is the thermal conductivity of medium.
3. Results and discussion
The behavior of nanoparticle concentration on temperature distributions in solid and fluid phases is displayed in section. For fluid phase, consider the nanofluid which is repaired by water and alumina oxide nanoparticles. The porous medium is taken as solid phase that is made by open-celled copper metallic foams. The governing equations for physical problem are demonstrated by taking Brinkman-LTNE/Brinkman-LTE and Darcy-LTNE/Darcy-LTE models and converted into non-dimensionless form to find its exact solution. The exact solutions of these equations are obtained by using computational software Mathematica 9. To see the effects of nanoparticle concentration, the values of other embedding parameters are taken to be fixed as ε=0.7, s=0.64, C=0.0015, and D=1.
The influence of nanoparticle concentration on the temperature profiles of fluid and solid phases for Brinkman-LTNE and Darcy-LTNE models is shown in Figure 2. It shows that the temperature profiles of fluid and solid are increased due to improvement in thermal physical properties of fluids especially thermal conductivity through increasing the concentrations of nanoparticles. Here negative sign shows that fluid transfers the heat to the wall. In this regard, temperature profiles of solid phase in both models are increased because of increasing the temperature of fluid phase by nanoparticle concentrations. The temperature variation for Brinkman-LTNE and Darcy-LTNE models has similar trend, but the temperature profile of Brinkman-LTNE models is found maximum as compared to the Darcy-LTNE models.
Figure 2.
Effect of nanoparticle concentrations on the temperature profiles of Brinkman-LTNE and Darcy-LTNE models.
The temperature profiles for Brinkman-LTE and Darcy-LTE models with effects of nanoparticle concentration are displayed in Figure 3. In Figure 3, it is seen that the temperature profiles for both models are amplified by increasing the nanoparticle concentrations. In comparison of models, it is noted that the effects of nanoparticle concentrations are dominant in the case of LTE as compared to LTNE. Moreover, the temperature at wall for LTE and LTNE models is the same but with maximum boost in the case of LTNE models at center. Moreover, the heat transfer with LTNE approach is smaller as compared to LTE approach because of thermal resistance due to solid phase.
Figure 3.
Effect of nanoparticle concentrations on the temperature profile of Brinkman-LTE and Darcy-LTE models.
4. Conclusions
A fully developed heat and mass flow of Al2O3-water nanofluid in a parallel-plate channel filled with porous media of Cu material is investigated by using Darcy-LTE/Darcy-LTNE and Brinkman-LTE/Brinkman-LTNE models. It is found that the distribution of temperature is improved in both approaches of heat transfer by using nanofluid. But temperature distribution is overestimated in LTE approach as compared to LTNE approach. This overestimate results are due to neglecting the difference between thermal conductivities of fluid and solid phase.
Nomenclature
u
velocity
μe
effective viscosity
μnf
viscosity of nanofluid
μf
viscosity of base fluid
ε
porosity
Tf
temperature of fluid
Ts
temperature of solid phase
Tw
temperature at interface
ρCp
heat capacity
ϕ
nanoparticle volume fraction
K
permeability
ke
effective thermal conductivity
km
thermal conductivity of medium
kfe
thermal conductivity of fluid phase
kse
thermal conductivity of solid phase
kp
thermal conductivity of nanoparticle
kf
thermal conductivity of base fluid
qw
heat flux
h
heat transfer coefficient
H
height
A
specific surface area
\n',keywords:"nanofluid, porous media, Darcy-LTE, Brinkman-LTE, Darcy-LTNE, Brinkman-LTNE models",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/65045.pdf",chapterXML:"https://mts.intechopen.com/source/xml/65045.xml",downloadPdfUrl:"/chapter/pdf-download/65045",previewPdfUrl:"/chapter/pdf-preview/65045",totalDownloads:756,totalViews:0,totalCrossrefCites:0,dateSubmitted:"September 10th 2018",dateReviewed:"December 11th 2018",datePrePublished:"September 27th 2019",datePublished:"August 19th 2020",dateFinished:"January 3rd 2019",readingETA:"0",abstract:"In current study, fully developed flow of Al2O3-water nanofluid with forced convection heat transfer in channel is investigated with different models of heat and mass distribution. The channel is filled with porous media of open-celled Cu metal foams. The Darcy and Brinkman models are used for the mass flow; however, the heat transfer distribution is examined through the local thermal equilibrium (LTE) and the local thermal non-equilibrium (LTNE) models. Exact solutions for Darcy-LTE, Brinkman-LTE, Darcy-LTNE, and Brinkman-LTNE models are obtained. Temperature profiles by these different models are discussed under effect of nanoparticle concentration and compare the profiles with each other.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/65045",risUrl:"/chapter/ris/65045",signatures:"Mohsan Hassan",book:{id:"7774",type:"book",title:"Nanofluid Flow in Porous Media",subtitle:null,fullTitle:"Nanofluid Flow in Porous Media",slug:"nanofluid-flow-in-porous-media",publishedDate:"August 19th 2020",bookSignature:"Mohsen Sheikholeslami Kandelousi, Sadia Ameen, M. Shaheer Akhtar and Hyung-Shik Shin",coverURL:"https://cdn.intechopen.com/books/images_new/7774.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",isbn:"978-1-78923-838-9",printIsbn:"978-1-78923-837-2",pdfIsbn:"978-1-83968-342-8",isAvailableForWebshopOrdering:!0,editors:[{id:"185811",title:"Dr.",name:"Mohsen",middleName:null,surname:"Sheikholeslami Kandelousi",slug:"mohsen-sheikholeslami-kandelousi",fullName:"Mohsen Sheikholeslami Kandelousi"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:null,sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Modeling and formulation",level:"1"},{id:"sec_2_2",title:"2.1 Darcy-LTNE model",level:"2"},{id:"sec_3_2",title:"2.2 Darcy-LTE model",level:"2"},{id:"sec_4_2",title:"2.3 Brinkman-LTNE model",level:"2"},{id:"sec_5_2",title:"2.4 Brinkman-LTE model",level:"2"},{id:"sec_6_2",title:"2.5 Physical properties",level:"2"},{id:"sec_8",title:"3. Results and discussion",level:"1"},{id:"sec_9",title:"4. Conclusions",level:"1"},{id:"sec_10",title:"Nomenclature",level:"1"}],chapterReferences:[{id:"B1",body:'Poulikakos D, Kazmierczak M. Forced convection in a duct partially filled with a porous material. Journal of Heat Transfer. 1987;109:653-662'},{id:"B2",body:'Vafai K, Kim SJ. Forced convection in a channel filled with a porous medium: An exact solution. Journal of Heat Transfer. 1989;111:1103-1106'},{id:"B3",body:'Gong W, Han J, Cheng F. Theoretical study of heat transfer enhancement in pipe with porous body. Journal of Hydrodynamics, Series B. 2001;13:111-116'},{id:"B4",body:'Cheng P, Hsu CT. Fully-developed, forced convective flow through an annular packed-sphere bed with wall effects. International Journal of Heat and Mass Transfer. 1986;29:1843-1853'},{id:"B5",body:'Mitrovic J, Maletic B. Effect of thermal asymmetry on laminar forced convection heat transfer in a porous annular channel. Chemical Engineering and Technology. 2006;29:750-760'},{id:"B6",body:'Sheikholeslami M, Shahzad SA, Li Z, Ahmad S. Numerical modeling for alumina nanofluid magnetohydrodynamic convective heat transfer in a permeable medium using Darcy law. International Journal of Heat and Mass Transfer. 2018;127:614-622'},{id:"B7",body:'Sheikholeslami M. CuO-water nanofluid free convection in a porous cavity considering Darcy law. European Physical Journal Plus. 2017;132:55-66'},{id:"B8",body:'Kuznetsov AV. Analysis of a non-thermal equilibrium fluid flow in a concentric tube annulus filled with a porous medium. International Communications in Heat and Mass Transfer. 1996;23:929-938'},{id:"B9",body:'Xu HJ, Gong L, Zhao CY, Yang YH, Xu ZG. Analytical considerations of local thermal non-equilibrium conditions for thermal transport in metal foams. International Journal of Thermal Sciences. 2015;95:73-87'},{id:"B10",body:'Lu W, Zhao CY, Tassou SA. Thermal analysis on metal-foam filled heat exchangers. Part I: Metal-foam filled pipes. International Journal of Heat and Mass Transfer. 2006;49:2751-2761'},{id:"B11",body:'Zhao CY, Lu W, Tassou SA. Thermal analysis on metal-foam filled heat exchangers. Part II: Tube heat exchangers. International Journal of Heat and Mass Transfer. 2006;49:2762-2770'},{id:"B12",body:'Ouyang XL, Vafai K, Jiang PX. Analysis of thermally developing flow in porous media under local thermal non-equilibrium conditions. International Journal of Heat and Mass Transfer. 2013;67:768-775'},{id:"B13",body:'Xu HJ, Zhao CY, Xu ZG. Analytical considerations of slip flow and heat transfer through microfoams in mini/microchannels with asymmetric wall heat fluxes. Applied Thermal Engineering. 2016;93:15-26'},{id:"B14",body:'Shaikh AW, Memon GQ. Analytical and numerical solutions of fluid flow filled with and without porous media in circular pipes. Applied Mathematics and Computation. 2014;232:983-999'},{id:"B15",body:'Sheikholeslami M, Houman BR. CVFEM for effect of Lorentz forces on nanofluid flow in a porous complex shaped enclosure by means of non-equilibrium model. Journal of Molecular Liquids. 2018;254:446-462'},{id:"B16",body:'Hassan M. Impact of iron oxide particles concentration under a highly oscillating magnetic field on ferrofluid flow. European Physical Journal Plus. 2018;133:230-244'},{id:"B17",body:'Hassan M, Marin M, Ellahi R, Alamri SZ. Exploration of convective heat transfer and flow characteristics synthesis by Cu-Ag/Water hybrid-nanofluids. Heat Transfer Research. 2018;49:1837-1848'},{id:"B18",body:'Hassan M, Marin M, Alsharif A, Ellahi R. Convective heat transfer flow of nanofluid in a porous medium over wavy surface. Physics Letters A. 2018;382:2749-2753'},{id:"B19",body:'Sheikholeslami M, Ellahi R, Hassan M, Soleimani A. A study of natural convection heat transfer in a nanofluid filled enclosure with elliptic inner cylinder. International Journal of Numerical Methods for Heat and Fluid Flow. 2014;24:1906-1927'},{id:"B20",body:'Sheikholeslami M, Ganji DD. Application of Nanofluid for Heat Transfer Enhancement. Elsevier Science; 2017. DOI: org/10.1016/B978-0-08-102172-9.00016-2'},{id:"B21",body:'Sheikholeslami M. Numerical approach for MHD Al2O3-water nanofluid transportation inside a permeable medium using innovative computer method. Computer Methods in Applied Mechanics and Engineering. 2019;344:306-318'},{id:"B22",body:'Sheikholeslami M. New computational approach for exergy and entropy analysis of nanofluid under the impact of Lorentz force through a porous media. Computer Methods in Applied Mechanics and Engineering. 2019;344:319-333'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"Mohsan Hassan",address:"mohsan.hassan@cuilahore.edu.pk",affiliation:'
Department of Mathematics, CUI, Lahore, Pakistan
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With the usual ball-and-socket design, patients obtain faster pain relief, stronger grip function, and shorter convalescence than with trapeziectomy. However, prostheses currently used have led to various early complications, especially in active and young patients. Revisions are most often due to the loosening of the trapezium cup and of the metacarpal stem and to the luxation of the first metacarpal bone. The short lifespan of these devices suggests the difficulty of designing a prosthesis that respects the complex anatomy and motions of the TMC joint. Early implant failure may reflect the fact that the current devices do not exactly replicate the true kinematics. The aim of this chapter will be to shed light mechanical explanations for TMC prosthesis failures in regard to the complex kinematics of the joint.",book:{id:"6843",slug:"biomechanics",title:"Biomechanics",fullTitle:"Biomechanics"},signatures:"Victoria Spartacus",authors:[{id:"256419",title:"Dr.",name:"Victoria",middleName:null,surname:"Spartacus",slug:"victoria-spartacus",fullName:"Victoria Spartacus"}]},{id:"19665",title:"Biomechanics of Competitive Swimming Strokes",slug:"biomechanics-of-competitive-swimming-strokes",totalDownloads:12997,totalCrossrefCites:0,totalDimensionsCites:16,abstract:null,book:{id:"917",slug:"biomechanics-in-applications",title:"Biomechanics in Applications",fullTitle:"Biomechanics in Applications"},signatures:"Tiago M. Barbosa, Daniel A. Marinho, Mário J. Costa and António J. 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