FTIR analysis data showing various functional groups present in dextran sulfate (DS) and AgNP‐DS complex.
\r\n\tCases of Corrosion in PA industrial equipment and plants are presented and discussed, based on the author's experience and knowledge.
\r\n\r\n\t
\r\n\tA singular application is the manufacture of artificial apatite for coating on stainless steel (SS) orthopedic implants in the human body.
Investigation of nanoparticles by different methods, especially by Fourier transform infrared (FTIR) spectroscopy, is very interesting in last years since they have a wide potential application in different industries [1–5]. Thus, silver nanoparticles (AgNP) of polysaccharide type and other natural products have scientific interest, but practical importance too, because of their application in pharmaceutical and cosmetic products development due to proven antimicrobial and antioxidant activities [6–9]. The nanoparticles are commonly synthesized by silver (or other metals) ions reduction to elementary state. But, reducing agents should also possess stabilizing properties in order to prevent aggregation [1]. Bankura and coworkers described a simple method of AgNP synthesis at room temperature from dextran, as well as their characterization and microbiological activity [6]. Pullulan‐mediated Ag nanoparticles, their synthesis, characterization, and microbiological activities are also reported [4]. Soluble starch, starch‐like polysaccharides, and chitosan are used in AgNP synthesis [5–9]. AgNP chitosan/gelatin bionanocomposites have also been studied [5]. Compounds containing carbonyl group are relatively easy complexed with different metals [10–12]. This functional group contains some polysaccharide derivatives like carboxymethyl cellulose (CMC) or carboxymethyl dextran (CMD), which are obtained by different chemical reactions of proton exchange between OH groups of glucoside moiety and carboxymethyl groups. AgNP‐CMC nanoparticles were prepared in weak alkaline solution by reaction of AgNO3 with CMC as a reducing and capping agent. It has been established that size distribution and morphology of mentioned nanoparticles are depended on Ag: CMC weight ratio, reaction time, temperature, and pH value of the reaction system. FTIR spectrophotometric analysis has shown that interactions between AgNP and polysaccharide have steric character [13]. Studies of composite hydrazine‐CMD and CMD magnetic Fe‐based nanoparticles [14, 15] have shown that solubility of these nanoparticles depends on pH value (NaOH), not on CMD content. This fact indicates on strong interactions of carbonyl group with magnetic nanoparticles on the surface.
\nHence, there are indications that carboxymethyl dextran form nanoparticles with Ag ions. CMD possess COOH group which can react with positively charged Ag ions to form complex compounds, but, it can also reduce Ag ions and stabilize the formed nanoparticles as in the case of hyaluronic acid with AgNP [16]. Since CMD has the ability to form nanoparticles, the intention of this chapter is to contribute in clarification of AgNP synthesis, physico‐chemical characterization by FTIR spectroscopic, diffraction and chromatographic methods, as well as testing of their antimicrobial activity. On the other hand, the biocomplexes based on exopolysaccharides are important in treatment of biometals deficiency in human and veterinary medicine [1, 17]. Polysaccharides, oligosaccharides and their derivatives, as well as simple sugars, may be used as ligands for the synthesis of biocomplexes with different metal ions (Cu, Fe, and Zn). These biocomplexes have an important role in metal ions transporting in organism [18]. Despite a number of studies of this kind of complexes, the investigations of effect of their structure to pharmaco‐biological activity are still interesting [19]. In this respect, the presented chapter offers further progress in the investigation of cobalt complex synthesis with dextran oligosaccharide, spectroscopic characterization, and the spectra‐structure correlation by various FTIR techniques.
\nRecent investigations of nanoparticles synthesis are mostly directed to green synthetic methods development. These methods include nontoxic reagents, synthesis procedures without problematic side products, and especially usage of biodegradable materials. Thus, chemical reduction in silver ions is the most frequently used besides photochemical or electrochemical methods [1]. In order to reduce and stabilize the Ag nanoparticles, the polysaccharides (dextran, starch, pullulan, cellulose) and their derivatives (dextran sulfate-DS, carboxymethyl cellulose, carboxymethyl dextran, chitosan, hyaluronic acid, heparin), and biocomposites AgNP chitosan/gelatin, are developed and improved [2–6, 13, 14, 16, 20–25]. Characterization of these particles has been carried out by UV‐Vis, FTIR spectroscopy, X‐ray diffraction (XRD), and energy‐dispersive X‐ray (EDX) methods. Electronic microscopy techniques [scanning electron microscopy (SEM) and TEM] are used for particle size determination and distribution, as well as shape defining. It is interesting that question of interaction nature between reducing and stabilizing agents with AgNP is still opened. Some authors [13] consider that steric physical interactions are relevant, while others [2, 16] give an explanation via coordination complex of Ag ions with reducing and stabilizing agents which contain suitable functional groups (COOH, NH2, OH, OSO3H), as in the case of Cu(II) ions complexes with carboxymethyl dextran or dextran sulfate (Figure 1).
\nStructural fragment of dextran sulfate sodium salt (DS) and carboxymethyl dextran (CMD) molecule.
Having in mind these facts, it can be assumed that dextran sulfate, which contains one or more sulfo groups in its structure, can be used as reducing and stabilizing agent for the AgNP synthesis. Also, it may be assumed that dextran sulfate forms complexes similar to CMD about what there are no literature data. Therefore, investigations in this chapter are related to the AgNP‐DS and AgNP‐CMD synthesis, their characterization by different methods and antimicrobial activity determination.
\nThe synthesis of AgNP‐DS has been performed in a reactor at temperature of 100°C, during 240 min, at constant pH of 7.5 and continuously stirring. Dextran sulfate has been used as a ligand in the synthesis. The synthesis is performed by DS solution (100 cm3, 0.002 M) adding in 100 cm3 of AgNO3 solution (0.001 M). The complex formation has been monitored via changing of reaction solution color, from white to yellow. The AgNP‐DS complex was precipitated with 96% ethanol after cooling down the reaction mixture to the room temperature. The obtained product has been dried at105°C under vacuum during 180 min. On the other hand, the carboxymethyl dextran has been used in the case of the AgNP‐CMD complex synthesis. The synthesis has been performed by 100 cm3 of AgNO3 (0.001 M) solution adding in 200 cm3 of CMD ligand solution (0.002 M) at constant pH of 7.0 (adjusted by NaOH). The synthesis is performed to the defined M:L ratio (from 1:1 to 1:2) by changing of reagents volume. The complexation has been performed at 100°C and continuously stirring during 120 min. A successful outcome of the AgNP‐CMD complex synthesis has been identified by changing of reaction solution color, from white to yellow. The reaction mixture has kept under reflux 24 h more, after that it has cooled to room temperature, and the complex AgNP‐CMD has precipitated by 96% ethanol. Final product has been dried at 105°C under vacuum during 180 min. The prepared AgNP‐DS and AgNP‐CMD complexes were characterized by different methods (FTIR, UV‐Vis, SEM, XRD, EDX) and by antimicrobial activity.
\nThe FTIR spectra were recorded by BOMEM MB‐100 (Hartmann & Braun, Canada) FTIR spectroscope and by KBr technique, at room temperature with 2 cm-1 resolution. Spectra‐structure correlation has been performed on empirical manner [2, 22], by comparing the spectra of ligands (DS and CMD) with the spectra of their complexes (AgNP‐DS and AgNP‐CMD). The appropriate FTIR spectra are shown in Figures 2 and 3.
\nFTIR spectra of dextran sulfate sodium salt (a) and AgNP‐DS complex (b).
FTIR spectra of dextran (a), CMD (b), and AgNP‐CMD (c).
The results of the complex AgNP‐DS spectral analysis are shown in Table 1. They show the position and assignations of bands that come from vibrations of all types of sulfo groups in DS and AgPN‐DS, as well as bands of deformation CH vibrations outside of the plane of glucopyranose unit which are characteristic for its conformation determining. As it can be seen in Table 1, there is a difference in the position of the νas(S–O) band which is shifted ~22 cm-1 toward lower frequencies in the AgPN‐DS spectrum (Figure 2), as well as the band from νas(O–S–O) which is shifted ~8 cm-1 toward higher frequencies. This difference in the bands position indicates the formation of coordination complexes between Ag ions and DS, where there is a change in conformation of sulfo groups from Eq to Ax position. The appearance of the spectrum in the area of C–CH out‐of‐plane deformational vibrations surface coupled with C–C–O, O–C–O and C–O–C vibrations depends on glucopyranose unit conformation [25–27]. When it is 4C1 conformation, the bands at ~915 cm-1 (weak), 850 cm-1 (shoulder), and 752 cm-1 are expected in the spectrum. The results from Table 1 show that starting DS retains the same conformation of glucopyranose unit during complex with Ag ions formation. There is a sharp intensive band at 1384 cm-1 in the FTIR spectrum of AgNP‐DS (Figure 2) observed by other authors who have investigated similar complexes and given the explanation of its origin [4, 5, 13, 22]. So, in the case of AgNP‐CMD complex (Figure 3c), some authors consider that this band is a result of νs(O–N–O) at O=NO2\n− radicals which are formed from AgNO3 agents participating in the formation of nanoparticles through the surface interactions [13].
\nAssignation | \nDS (cm-1) | \nAgNP‐DS (cm-1) | \nΔν (cm-1) | \n
---|---|---|---|
νas (S-O) | \n1261 | \n1239 | \n22 | \n
νs (S-O) | \n988 | \n1060 | \n72 | \n
νas (O-S-O) | \n824 | \n832 | \n8 | \n
νs (O-S-O) | \n585 | \n588 | \n3 | \n
4C1 conformation of the α‐ | \n915 | \n915 | \n– | \n
850 | \n850 | \n– | \n|
745 | \n752 | \n7 | \n
FTIR analysis data showing various functional groups present in dextran sulfate (DS) and AgNP‐DS complex.
The infrared spectra of AgNP‐CMD products and starting CMD agent are compared with literature data and dextran spectrum because of the major bands assignment (Figure 3). In the CMD spectrum, vibrations of carboxymethyl groups: ν(C–O) around 1740 cm-1; deformation vibration δ(C–OH) which appears around 1250 cm-1; ν(C–O) vibration around 1150 cm-1; and deformation δ(C–O) vibration around 680 cm-1 are expected to oppose starting dextran for CMD synthesis. Stretching ν(C–O) vibration has been found in similar carboxymethyl polysaccharides; after carboxymethylation of the k‐carrageenan, ν(C–O) has found at 1737 cm-1 [28], for carboxymethylated glucan at 1736 cm-1 [29], as well as at 1750 cm-1 in the spectrum of CMD [14]. As it can be seen from Figure 3b, the CMD spectra possess bands at 1740, 1244, 1139, and 682 cm-1 (which are marked by arrows) from CO carboxymethyl vibration of all types. The aforementioned bands are not in the range of dextran (Figure 3a) as it is expected. Changes in the position of the above‐mentioned bands can be a good indicator of bonds type that is eventually formed by interaction with Ag+ ions [16]. Also, the changes in the area of deformation vibration of C–OH are expected. In the case of AgNP‐CMD coordination complexes formation, the frequency ν(C–O) band should be lower, or, if both O atoms of COOH groups participate in the coordination, the frequency of ν(C–O) vibration should be higher because of electron delocalization, as well as the absence of δ (C–OH) bands.
\nSpeaking about carboxylate anion, delocalization of electrons causes the order of two CO bonds to be the same, so two bands (at 1600 and 1400 cm-1) are expected in CO stretching vibration region, which are ascribed to asymmetric and symmetric C–O vibration, as it is indicated in the literature [13, 16, 28]. The similar situation is in AgNP‐CMD complex (Figure 3c) in the CO groups vibration area. In fact, in this area of the spectrum, there are two intensive bands (at 1603 and 1420 cm-1) which, according to its position and intensity should be attributed, νas(C–O) and νs(C–O) vibration, indicating coordination of Ag+ ions with a COOH group. In support of this is the absence of ν(C–O) and δ(C–OH) vibration bands. The appearance of spectrum in the area of 1000–700 cm-1, in all three tested compounds (Figure 3), is very similar and according to the literature data [30, 31] suggests 4C1 conformation of the glucopyranose unit.
\nAbsorption spectra of starting ligand compounds (DSi CMD) as well as of final complexes (AgNP‐DS and AgNP‐CMD) are obtained by UV‐Vis spectrophotometer (Varian Cary‐100 Conc.). Spectrophotometric analysis was carried out in the range of 200–800 nm using original Cary UV‐Conc. (Varian) software. The obtained UV‐Vis spectra are presented in Figures 4 and 5.
\nUV‐Vis spectra of dextran sulfate (DS) and AgNP‐DS complex in function of the synthesis time.
UV‐Vis spectra of CMD ligand, AgNP‐CMD = 1:1 complex (A), AgNP‐CMD = 1:2 complex (B), and AgNP‐CMD complex after 3 months (C).
Change in color from yellowish to brown, as well as careful interpretation of UV‐Vis spectra, is used for estimating of AgNP synthesis [2–10]. A strong absorption band, called SPR band (surface plasmon resonance), is expected in 370-450 nm region in the UV‐Vis spectrum of AgNP [32]. Its exact position depends on numerous factors (the most on AgNP size), while intensity depends on their concentration [4, 21, 33]. Changes in this band position are used as a criterion of the AgNP stability, that is, aggregation of the nanoparticles during the time. As it can be seen from Figure 4, the existence of SPR band at 410 nm indicates the AgNP formation. However, its position is changed during synthesis with time, but after 2 h remains constant at 420 nm. The estimated particle size of AgNP‐DS based on the UV data [34] is approximately 40 nm. A change in color from yellowish to brown during the synthesis has been observed in the case of AgNP‐CMD formation. SPR band for this complex (Figure 5), synthesized at different molar ratio, is located at 420 nm, which is not present in the starting CMD. It is characteristic that intensity of this band is proportional to the amount of AgPN‐CMD particles; it increases with increasing amounts of CMD, or during staying of the reaction mixture for 3 months (Figure 5C), which is similar to other studies [21]. Unchanged position of SPR band speaks in favor of good aggregation stability of synthesized particles. UV area below 300 nm was not investigated in the literature. However, in the UV spectra of tested compounds (Figure 5), there is an intense band of the formed complex at 215 nm (π→π* transition of the carboxyl group [35]) indicating red shift effect compared to CMD. This phenomenon is an indicator of Ag ions interaction with CMD and AgNP‐CMD complex formation.
\nCrystal structure of AgNP‐DS and AgNP‐CMD nanoparticles was determined and confirmed by X‐ray diffraction (XRD) technique. The samples were prepared by press and pull method in top‐loading specimen plate [36]. The diffractogram was measured in Bragg‐Brentano θ: 2θ geometry by a conventional powder diffractometer, Seifert V‐14, using Cu Kα radiation (λCu Kα1 = 1.5406 Å, Ni filter, generator settings: 30 kV, 30 mA). As an external standard for peak position calibration and instrumental peak broadening determination, LaB6 was used. XRD data were collected over the 2θrange of 5–90°with a step size of 0.02°, and an exposition time of 2 s per step. The obtained diffractograms are shown in Figures 6 and 7.
\nXRD diffraction patterns of AgNP‐DS complex.
XRD diffraction patterns of CMD and AgNP‐CMD complex.
From the presented X‐ray diffraction patterns of AgNP‐DS (Figure 6) can be noticed the XRD peaks at 38.24, 44.32, 64.58, 77.59,and 81.79°. Based on literature data [4, 6], the characteristic XRD peaks could be determined as next crystallographic planes: 111, 200, 220, 311, and 222. These planes are specific for the face‐centered cubic silver crystals. This statement, along with the specified values, indicates the presence of silver nanoparticles in the synthesized AgNP‐DS complex. Similar to the previous study, the crystal structure of Ag nanoparticles was determined with complex AgNP‐CMD. Based on X‐ray diffraction patterns (Figure 7) and the presence of XRD peaks at 38.02, 44.50, and 64.51°, a particular crystallographic planes are as follows: 111, 200, and 220, which are specific for the cubic silver crystals. According to literature [4, 6], the XRD peak at 29.01° is characteristic of the CMD ligand.
\nThe calculation of average AgNP size has been done from the width of reflection in the X‐ray diffraction pattern according to the Scherrer\'s equation (1):
\nwhere D is the mean size of metal nanoparticles (nm); K is Scherrer constant (it\'s chosen 0.9—roughly spherical particles); λ is wavelength of X‐ray radiation (nm);θ is angle of diffraction (°); and FWS\n is specimen broadening of single peak (in radians). FWS\nis obtained according to the Eq. (2):
\nwhere FWHM is full width at half maximum of the peak; FWI\nis instrumental broadening gained from LaB6 diffractogram at the similar 2θangles; and d is parameter of deconvolution (here d is chosen as 1.5 which means that shape is partly Gaussian and partly Lorentzian). According to Scherrer\'s equation (1) and XRD peak at 38.24° 2θ from diffraction patterns (Figure 7), it is concluded that AgNP have mean crystallite size of 40 ± 4 nm.
\nThe size and shape of AgNP‐DS and AgNP‐CMD complexes were further characterized by scanning electron microscopy (SEM) on JEOL JSM 5300 scanning electron microscope. Scanning micrographs were transformed into a PC format in order to further analyze the particles morphology. The samples for SEM analysis have been prepared by thin layer of the complex suspension overnight air drying at room temperature. Dried samples have been coated with 10‐nm‐thick film of gold in JPC JEOL‐1100 apparatus. Electron beam of 30 keV has been used. The SEM micrographs of AgNP‐DS (Figure 8A) showed both individual particles, but a number of aggregates, too. Size of 10–60 nm is predominant for individual spherical particles. Images have also indicated that obtained nanoparticles are stable, and they are not in a mutual contact. This can be ascribed to stabilization of the nanoparticles by DS as a capping agent. Aggregates of nanoparticles with poorly defined morphology and irregular structure have also been found (Figure 8A).
\nSEM images of AgNP‐DS showing the existence of individual nanoparticle and large aggregates (A) and EDX spectrum of individual AgNP‐DS (B.)
Energy‐dispersive X‐ray (EDX) spectral analysis has been performed by LINK Analytical 2000 QX microprobe assembled on a JEOL JSM 5300 scanning electron microscope. Samples prepared for SEM analyses have been used for EDX spectra measuring. EDX spectroscopy can be used for qualitative as well as quantitative assessment of silver used for the AgNP production [36]. EDX spectrum of AgNP‐DS is shown in Figure 8B. Strong signal comes from elemental silver, while weaker signals come from S, O, and Na (from Na salt of DS), confirming that AgNP are formed as a part of AgNP‐DS. This is consistent with an optical absorption peak appearance at approximately 3 eV (410 nm), which originates from SPR, and it is characteristic for metallic silver nanocrystals [6].
\nSimilar to the previous complex, the SEM micrographs of AgNP‐CMD (Figure 9A) show single particles, but a number of aggregates as well. Particle size of 10–60 nm is dominant for individual spherical particles. SEM images showed that obtained nanoparticles are stable and not in direct contact with each other. This can be explained as stabilization effect of CMD, as a capping agent, on produced nanoparticles. But, aggregated nanoparticles with larger irregular structure and no well‐defined morphology were also found (Figure 9B).
\nSEM images of individual spherical particles (A) and aggregated nanoparticles of AgNP‐CMD (B).
Agar disk diffusion method has been used for measuring of antibacterial and antifungal activity of AgNP stabilized by DS. One fungal strain (Candida albicans ATTC 2091) and nine bacterial strains such as Gram‐positive (Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 11778, Bacillus luteusin haus strain, Bacillus subtilis ATTC 6633, and Listeria monocytogenes ATCC 15313) and Gram‐negative (Escherichia coli ATTC 25922, Pseudomonas aeruginosa ATTC 27853, Klebsiella pneumoniae ATTC 700603, and Proteus vulgaris ATTC 8427) were used as an indicator strain for this analysis. Preparation of suspension was performed by already described method [37]. Direct colony method has been used for bacterial and yeast suspensions preparation, and the colonies have been taken directly from the plate and suspended in 5 cm3 of sterile 0.85% saline. Turbidity of the initial suspension has been adjusted comparing with 0.5 McFarland\'s [38]. After this adjustment, the bacterium and yeast suspensions contained close to 108 and 106 colony‐forming units (CFU)/cm3, respectively. Initial suspension has been additionally prepared by tenfold dilution into sterile 0.85% saline. Inoculation of bacterial cell suspensions has been done to the trypton soya agar plates, while the yeast suspension to the Sabouraud maltose agar plates. Standard sterile cellulose disks of 9 mm diameter have been impregnated with different AgNP‐DS concentrations (0.25, 0.5, 1.0 mg cm-3) and putted on surface of the inoculated plates. The plates have been incubated at 37°C for 24 h. Inhibition zones were evaluated by measuring the diameter of the zones growth (Table 2).
\nMicrobes | \nAgNP‐DS concentration | \n|||
---|---|---|---|---|
0.25 mg cm-3 | \n0.5 mg cm-3 | \n1.0 mg cm-3 | \n||
Fungi | \nC. albicans | \n– | \n16 | \n– | \n
Bacteria G+ | \nL. monocytogenes | \n16 | \n17 | \n18 | \n
B. cereus | \n16 | \n18 | \n19 | \n|
B. subtilis | \n16 | \n17 | \n19 | \n|
S. aureus | \n17 | \n18 | \n19 | \n|
B. luteus haus strain | \n20 | \n21 | \n24 | \n|
BacteriaG- | \nP. vulgaris | \n13 | \n14 | \n15 | \n
K. pneumoniae | \n16 | \n18 | \n19 | \n|
E. coli | \n17 | \n18 | \n21 | \n|
P. aeruginosa | \n23 | \n24 | \n26 | \n
Antimicrobial activity of AgNP‐DS, radial diameter of inhibition zones (mm) for tested bacterial and fungal strains.
The investigated AgNP‐DS solution has shown antibacterial activity against S. aureus, B. cereus, B. luteus in haus strain, B. subtilis, L. monocytogenes, E. coli, P. aeruginosa, K. pneumoniae, and P. vulgaris bacteria, which is proved by clear inhibition zones of the bacteria growth around the disks (Table 2). Inhibition has been observed for all analyzed bacterial strains in the 0.25 mg cm-3 concentration of AgNP‐DS, indicating relatively low minimal inhibitory concentration against these microorganisms. For example, Dhand and coworkers [39] stated that minimal inhibitory concentrations for E. coli and S. aureuss were around 0.26 mg cm-3. The highest inhibition zones were observed against P. aeruginosa and B. luteusin haus strain, and inhibition zones of AgNP‐DS against these microorganisms in 1.0 mg cm-3 concentration were 26 and 24 mm, respectively. P. vulgaris was the least sensitive to the AgNP‐DS (1.0 mg cm-3) activity with zone of 15 mm. Investigation of AgNP‐DS activity in different concentrations against other bacterial strains has shown similar results with inhibition zones of 16–17 mm, 18–19 mm, and 18–21 mm for the AgNP‐DS concentration of 0.25, 0.5, and 1.0 mg cm-3, respectively. The results for K. pneumoniae, B. luteus in haus strain, and P. aeruginosa are higher compared to data for AgNP‐CMD (Table 3). Antifungal activity against C. albicans was observed only in the concentration of 0.5 mg cm-3 AgNP‐DS. Low antimicrobial activity of AgNP against C. albicans has been estimated for AgNP stabilized with CMD. The mechanism of AgNP antimicrobial activity can be related to silver accumulation in the membranes of bacteria, which cause cell death [40]. Silver cation can react with thiol groups and proteins in the cells; nonetheless, it can inactivate enzymes essential for the normal cell metabolism [41]. The investigated AgNP‐DS particles, in the concentration of 1.0 mg cm-3, have shown a number of specificity concerning its antimicrobial activity. It is important that higher concentration of silver is harmful for consumer and for microbes as well, so the lower concentrations are much more applicable. The effective concentrations of AgNP, which have effect in organisms different from the control, are in the range from a few ng dm-3 to 10 mg dm-3; this effective concentration is depended on the organism itself as well as many other factors [42]. Having in mind these results, it can be concluded that this design of silver nanoparticles synthesis has a great potential because of their antimicrobial activity.
\nMicrobes | \nAgNP‐CMD concentration | \n|||
---|---|---|---|---|
0.25 mg cm-3 | \n0.5 mg cm-3 | \n1.0 mg cm-3 | \n||
Bacteria G+ | \nB. lutea | \n11 | \n13 | \n20 | \n
\n | B. cereus | \n11 | \n12 | \n14 | \n
\n | B. aureus | \n12 | \n18 | \n21 | \n
Bacteria G- | \nE. fecalis | \n– | \n– | \n11 | \n
\n | P. aeruginosa | \n– | \n– | \n12 | \n
\n | K. pneumoniae | \n13 | \n14 | \n15 | \n
Fungi | \nAspergillus spp. | \n– | \n– | \n12 | \n
\n | Penicillium spp. | \n13 | \n20 | \n20 | \n
\n | C. albicans | \n– | \n– | \n11 | \n
Antimicrobial activity of AgNP‐CMD, radial diameter of inhibitionzones (mm) for tested bacterial and fungal strains.
In order to compare antimicrobial activity of similar complexes, the results of AgNP‐CMD antimicrobial activity (radial diameter of inhibition zones) are presented in Table 3. The AgNP‐CMD solution exhibited antibacterial activity against bacteria B. lutea, B. aureus, B. cereus, E. fecalis, P. aeruginosa, and K. pneumoniae showing clear inhibition zones of the bacteria growth around the disk. AgNP‐CMD in the concentration of 1.0 mg cm-3 have shown a number of specificity concerning its antimicrobial activity. The antifungal activity of the AgNP‐CMD has been analyzed by agar disk diffusion method. Aspergillus spp., Penicillium spp., and C. albicans were inhibited in a concentration‐dependent manner. The radial growth inhibition zones increased with the AgNP‐CMD concentration increasing from 0.25 to 1.0 mg cm-3. The fungus Penicillium spp. was more sensitive to the AgNP‐CMD comparing to the other two fungal strains.
\nA lot of investigations in the field of coordination chemistry are based on synthesis and characterizations of different biocomplexes present in the biological systems. Synthetic ligands, which can serve as model molecules for complex biomolecular structures, are also investigated [19]. Bioligands or synthetic ligands are mostly natural macromolecular compounds. These products of special importance mostly represent complexes of different metals (Fe, Co, Cu, Zn) with ligands of polysaccharide type (such as pullulan, inulin, dextran) [43–45]. However, the native polysaccharide possessing antigen characteristics wherefore is not of pharmaceutical importance [18]. Depolymerization of raw polysaccharides, trying to get products with adequate molar masses distribution for commercial purposes, has been done. Dextran, is a well‐known, extracellular, water‐soluble neutral polysaccharide with α‐(1–6)‐linked
The cobalt complexes with reduced low‐molar dextran as ligand (Co(II)‐RLMD) were synthesized in water solutions, at different pH values (7.5-13.5) and different temperatures (298–373 K), using CoCl2 × 6H2O and RLMD (5000 g/mol). The details of synthesis have been described [54, 55]. The complexes were isolated in the solid state. For further structural examination, the samples of Co(II)–RLMD were deuterated (D2O, Merck) for 2 h, at room temperature, in vacuum.
\nKBr pastille method has been used for sample preparation. The FTIR spectra have been recorded at room temperature, as an average value of 40 scans (resolution of 2 cm-1) on a Bomem MB‐100 FTIR spectrometer (Hartmann & Braun, Canada) coupled with a DTGS/KBr detector. Spectra‐structural correlation of dextran by FTIR spectroscopy has been the subject of attention of many researchers [30, 31, 56–58]. It was shown that by studying the individual spectral areas, the information on linearity [content of α‐(1–6) bond], crystallinity, conformation, conformational transitions, and changes in the structure of differently treated dextrans can be obtained. The FTIR spectrum of reduced low‐molar dextran is shown in Figure 11a. Bands at 765 and 916 cm-1 are indicating the presence of the α‐(1–6) glycosidic bonds, and the estimated content of these bonds is greater than 96% that indicates a high linearity of polysaccharide. The presence of these bands as those at the 845 cm-1 indicates a C1 conformation of glucopyranose units (eq‐ax‐ax‐ax‐ax arrangement of adjacent C–H groups). There is an intense broad band whose centroid is at about 3400 cm-1, in the area of stretching OH vibrations. Summary intensity of this band comes from the ν(O-H) vibrations of hydroxyl groups involved in the formation of several by the strength of hydrogen bonds, but also from the H2O molecule whose presence is confirmed by the band at 1640 cm-1, which is result of δ(HOH) vibrations [54].
\nThe FTIR spectra of RLMD (a) and Co(II)‐RLMD complexes synthesized on the boiling temperature and different pH: 7.5 (b) and 13.0 (c).
FTIR spectral segments of dextran (a), partially deuterated (b), and fullydeuterated (c) RLMD analogs in ν(O-H), ν(O-D), and δ(HOH) vibrations.
The FTIR spectra of synthesized Co(II)‐RLMD complex, which were obtained under various reaction conditions, are presented in Figure 10. The FTIR spectra of RLMD and its Co(II)‐RLMD complex are basically similar. In the FTIR spectra of synthesized complex, there are the differences in the area of O–H vibrations. In this area, there is a large, complex band approximately 3390 cm-1 of ν(O–H), which is likely due to the stretching vibrations of polysaccharide OH groups. The characteristic IR band of δ(HOH) at about 1645 cm-1 in the spectra of synthesized complexes, as well as in the spectrum of RLMD, as noted above, indicates the presence of crystal water in the structure [54, 55]. By analyzing the low‐frequency part of the RLMD spectrum (γ(C-H) vibrations, Figure 10a), the FTIR spectra of Co(II)‐RLMD complex (Figure 10b and c), and the presence of bands at about 915 and 845 cm-1, 4C1 conformation of glucopyranose units, which indicates that the complexation with Co(II) ions does not lead to conformational changes in glucopyranose units, can be determined. In accordance with this is the change in the intensity of IR band in the area approximately 1350 cm-1 that originates from δ(C-H) and δ(O-H) vibrations. With an increase in the pH synthesis (from 7.5 to 13.5), the band intensity of ν(O-H) vibration increases, and the frequency of ν(O-H) vibration band at lower pH (7.5–8.5) stays almost unchanged and then increases with increasing pH (11–13.5). If the complexation with Co(II) ions takes place via OH groups at the C–2, C–3, or C–4 carbon atoms of dextran glucopyranose units (involved in the formation of various by the strength of hydrogen bonds in dextran), hydrogen bonds disappear by complexation, so the bands are expected at the higher frequencies. In complexes with the highest metal content (10.07% Co), which were synthesized at pH 12, a set of IR bands in this area is close to that at starting RLMD. In the complex which was synthesized at pH 13 with a minimum cobalt content of 1.89% in the IR spectrum (Figure 10c), there are intense bands at 3400 cm-1 to the binder of low‐frequency side in this region. This could indicate that the structure of this complex differs slightly from the structure of other Co(II)‐RLMD complex, which were synthesized under different reaction conditions. In the low‐frequency area (<800 cm-1) of the FTIR spectra of RLMD and Co(II)‐RLMD complex, there are some differences. In this region of the IR spectrum, in addition to the band of ν(Co-O), the bands of deformation γ(O-H) vibrations of polysaccharide as well as the deformation vibrations of glucopyranose ring are expected (Figure 10). Wide band of medium intensity in the FTIR spectra of Co(II)‐RLMD complex at about 450 cm-1 shows a fine structure.
\nIsotopic substitution of hydrogen atoms by deuterium, connected with FTIR spectroscopy, has an important role in determining the structure of dextran. Isotopic exchange results indicate that dextran and its Co(II)‐RLMD complex are crystal hydrates (probably one type of water molecules) [59]. Structural changes in the process have been detected by absorption bands in the area of 3600–3000 cm-1, caused by ν(O-H) vibrations. In the case of isotopic exchanges of O-H to O-D group, the frequency of stretching vibration is reduced to √2, and it is located in the area of 2700–2300 cm-1. Deuteration is a very sensitive method to assess the environment of OH groups, which is associated with the intensity generated by hydrogen bonds. The degree of crystallinity of the polysaccharide can be determined by FTIR spectroscopy method with deuteration. Crystallinity is a part of the regulated saccharide area in which the macromolecules are connected with parallel hydrogen bonds. In processing the sample with D2O, usually OH groups in less regulated or amorphous regions were rapidly converted into OD groups. Conversion of OH groups in the crystal areas is very slow. Thus, the degree of crystallinity has been determined by the change in intensity of asymmetrical ν(O-H) band vibrations and by the appearance of new bands of ν(O-D) vibration. The relations of band intensity at 1429 and 893 cm-1 were taken as empirical indicators of the degree of crystallinity of samples. With decrease in the crystallinity, the band at about 1430 cm-1 disappears and comes to an increase in the intensity of the band at approximately 900 cm-1, typical for the amorphousness. Even better relationship can be seen at the band at approximately 1370 and 2900 cm-1. Namely, in the spectrum of partially deuterated analogs of dextran (Figure 11) in the ν(O-D) area of the vibration of HDO molecules, there is a single band at about 2495 cm-1. Partners of these vibrations would be expected at about 3400 cm-1 in the ν(O–H) areas (taking into account the displacement factor of 1.35).
\nThe FTIR spectra of Co(II)‐RLMD complex (a) and its deuterated analog (b), which was synthesized at pH 13, are shown in Figure 12. In the FTIR spectrum of Co(II)‐RLMD complex (Figure 12b), in the area of ν(O-D) vibrations of HDO molecules, there is a single band at about 2483 cm-1 in the corresponding complexes with crossfold on the high‐frequency side. Partners of these vibrations would be expected at about 3400 cm-1 in ν(O-H) area. Results of partial deuteration indicate that the band at about 3400 cm-1 is sensitive to isotopic substitution, in both cases (RLMD and Co(II)‐RLMD complexes). Reducing the intensity of this band by deuteration demonstrates that the ν(O-H) vibrations of water molecules are its part. This fact indicates that both compounds contain crystal water in their structure. Confirmation of this conclusion is that in the spectra of deuterated analogs of both compounds (Figures 11 and 12), an intense band near 1645 cm-1 is also highly sensitive to isotopic substitution and is to be attributed to the HOH deformation vibration of the crystal water.
\nFTIR spectra of Co(II)‐RLMD complex (a) and its deuterated analog (b) synthesized at pH 13.
As known, Seidl et al. [60] proposed criteria according to which, based on the study of spectra of protonated, partially and fully deuterated hydrate, it is possible to determine the number of types of H2O molecules (n) and the number of nonequivalent OH groups (m). By the spectra appearances, in the stretching OD area of HDO molecules, and the appearance of a band, whose intensity increases monotonically with increasing degree of deuteration when the frequency does not change, it can be concluded from the above criteria [60] that in the structure of dextran and its complexes with Co(II) ions is present one crystallographic type of water molecule (n = 1). On the basis of Berglund correlation [61], from equation (3), Ow…O distances are estimated at 283.1 pm for dextran and 281.8 pm for Co(II)‐RLMD complex:
\nWater protons are involved in the formation of relatively weak hydrogen bonds (m = 1). In bending area of HDO and D2O, in the spectrum of deuterated analogs of the complex, there are bands around 1315 and 1070 cm-1, which confirm the previously disclosed consideration of the water binding. From FTIR spectrum shown in Figure 12b, decrease in the intensity of the band around 1430 cm-1 and an increase in the band intensity at approximately 910 cm-1 can be observed, which is a characteristic for amorphous character. An even better relationship can be observed in the FTIR spectrum of Figure 12, with the band of about 1370 and 2900 cm-1. Based on the results of FTIR spectroscopy, an amorphous structure of the synthesized Co(II)‐RLMD complex can be assumed.
\nThe ATR‐FTIR spectral analysis has been performed by microspectroscopy ATR‐FTIR system (Bruker, Tensor‐27). Within this system, FTIR spectroscope is connected to a microscope (15× objective) (Bruker, Hyperion‐1000/2000) and a computer system capable of microanalysis by using a liquid‐nitrogen‐cooled (250 μm) MCT detector (GMBH, Germany). The ATR‐FTIR spectra (Kubelka‐Munk option) have been recorded in the range of 4000–400 cm-1, with 4 cm-1 resolution and 260 scans. The newly formed FTIR vibrational microspectroscopy can provide information on the sample at the molecular level, with high spatial resolution at the microscopic level. Small sample can be analyzed by both nondestructive vibrational spectroscopic techniques (Raman, IR) [62–67]. Spectra can be recorded continuously in different parts of the microsample in order to obtain appropriate databases. Figures 13 and 14 show the absorption ATR‐FTIR spectra of Co(II)‐RLMD complex, which were obtained under various reaction conditions.
\nATR‐FTIR spectra of Co(II)‐RLMD complex synthesizedat the boiling temperature and pH values in the range of 7–11.
ATR‐FTIR spectra of Co(II)‐RLMD complex synthesized at the boiling temperature and pH values in the range of 12–14.
The wavenumber values of characteristic IR bands in the ATR‐FTIR spectra of Co(II)‐RLMD complex are given in Table 4.
\nWavenumber (cm-1) | \nBand assignation | \nIntensity | \nComment | \n
---|---|---|---|
3400 | \nν(O-H) | \nVery strong, complex | \nCH-OH glucopyranose, H2O | \n
2930 | \nνas(C-H) i νsy(C-H) | \nMedium | \nCH | \n
1640 | \nδ(HOH) | \nMedium | \nH2O | \n
1450–1345 | \nδ(C-H) | \nMedium | \nCH | \n
1420 | \nδ(O-H) | \nMedium | \nOH | \n
1150, 1110, 1070, 1040, and 1010 | \nνas(C-O), (C-O-C), νas(C-C-C) i (C-C-O) | \nVery strong, strong | \nGlucopyranose | \n
1000–700 | \nγ(C-H) | \nMedium | \nConfiguration | \n
Assignment of characteristic IR bands of RLMD and the synthesized Co(II)‐RLMD complexes.
Absorption bands corresponding to the specific chemical components can be represented as a map. ATR‐FTIR spectra, presented in Figures 13 and 14, correspond to the different parts of the sample of Co(II)‐RLMD complex, which show a homogeneity of the samples. A new way of visualization shows the capability of visualization not only of heterogeneous region of the samples, but also at the same time provides microspectroscopic spatial information. The visualization of different concentrations of components and presentation as 3D maps is also enabled. Application of ATR‐FTIR microscopy to Co(II)‐RLMD complex, which were synthesized under different reaction conditions, is shown in Figure 15. The changes in color contours at certain parts of the image indicate the content and distribution of cobalt and polysaccharides in Co(II)‐RLMD samples. ATR‐FTIR microspectroscopic data show a high homogeneity of the samples, and the presence of Co(II) ions (the results obtained by other spectroscopic techniques) has been confirmed by the color of Co(II)‐RLMD complex.
\nDifferent FTIR microscopic profiles (A–C) (300 μm × 250 μm) for ligand (RLMD) and Co(II)‐RLMD complexes at different pH values.
The modern Fourier transform infrared spectroscopic techniques (linear scan, reflection, transmission, mapping, video analysis) in combination with diffraction (XRD), energy‐dispersive X‐ray (EDX), spectrophotometric (UV‐Vis), and electronic microscopy (SEM) methods are applied in the structure analysis of synthesized green nanoparticles and polysaccharide complexes, as well as for the confirmation of suggested types of complexes structure and for the testing of samples homogeneity. In this respect, silver nanoparticles were prepared with dextran sulfate or carboxymethyl dextran as a reducing and capping agent, while cobalt biocomplexes were synthesized with reduced low‐molar dextran as ligand. Comparison of FTIR spectra of initial exopolysaccharide compounds (DS, CMD, RLMD) and final products (AgNP‐DS, AgNP‐CMD, Co(II)‐RLMD complexes), in the specific region of characteristic functional group vibrations, has indicated on coordination complexes forming as a part of complex structure. FTIR spectroscopic analysis has shown that interactions between metal ions and specific polysaccharide functional groups have steric character and suggest 4C1 conformation of the glucopyranose unit. The existence of nanoparticles (in range of 10–60 nm) has been confirmed by SPR band in the UV‐Vis spectra, by SEM microscopy, and XRD methods. AgNP size was determined on the Bragg reflection at 38.24°2θ, yielding mean crystallite size of 40 ± 4 nm. It has been found that crystalline structures of silver complexes are face‐centered cubic type by XRD method. Morphological SEM analysis has been shown that formed nanoparticles are spherical and inclined to aggregation. It has been established that size distribution and morphology of mentioned nanoparticles (by SEM and FTIR microspectroscopy methods), as well as the structural form of the complexes (by FTIR, UV‐Vis, XRD), are depended on ligand properties (such as constitution, degree of amorphousness or crystallinity, molar mass, units conformation, chain linearity) and on the reaction conditions (such as metal‐ligand weight ratio, reaction time, temperature, and pH values). Also, antimicrobial and antifungal activities of synthesized AgNP have been determined. The highest inhibition zones were observed against P. aeruginosa and B. luteusin haus strain, while P. vulgaris was the least sensitive to the nanoparticles. The fungus Penicillium spp. was more sensitive to the AgNP comparing to the other two fungal strains. Having in mind these results, it can be concluded that this design of silver nanoparticles synthesis has a great potential because of their antimicrobial activity.
\nThis study is the result of the project TR‐34012, funded by the Ministry of Education, Science and Technological Development, of the Republic of Serbia.
\nEpilepsy is a global health challenge, one that is responsible for a social and economic burden worldwide, it is estimated to be twice as common in low-income countries than that in the high-income countries, especially in a poor country like Sudan, resulting in unfair treatment, prejudice and stigma [1], and overwhelming decrease in quality of life [2]. People with epilepsy (PWE) in Sudan suffer from a collapsing and deficient health care system, and a community falling behind and lacking enough understanding towards their affliction, with a cultural heritage and misconceptions, and an educational system contributing to make it only that much more difficult for (PWE) to live a normal life, sometimes weighing them down and preventing them from seeking professional medical help altogether. The resultant treatment gab causes a mortality rate dwarfing that of first world countries [3]. On top of that, Sudan is lacking sufficient research and infrastructure to develop satisfying estimates about the situation on the ground, and recent data are scarce [3].
Sudan is the third largest country in Africa that occupies almost 728,000 square miles of northeast Africa. It sits along the sub-Saharan crossroads and along the cost of the red sea that runs through its east-northern borders. In addition to Egypt, Sudan shares borders with six other countries, which are Ethiopia, chad, Libya, Eritrea, Central African Republic, and lastly South Sudan that had its secession from Sudan by July 2011. Sudan is mainly formed of flat plains interspersed by mountain ranges, and due to its immense area, Sudan has different climates and several rivers coursing through the country, mainly the blue and white Niles that join together to form the river Nile in Khartoum the capital city of Sudan.
Although it’s an enormously sized country, it is sparsely populated compared to some of the African countries as it has an estimated census of 43 million people, the majority of which are rural in comparison to the urban population that is mainly centered in the capital.
Sudan is vastly enriched with different races, cultures and a blend of Arabic tribes that form the majority of the population and various African tribes and ethnicities, this enrichment may be contributing to its ever astounding cultural diversities and perhaps the fuel to political differences and the rather devastating civil wars that have crushed the country for ages, viciously affecting Sudan in every aspect possible. Sudanese people are still facing major challenges in everyday aspect of life duet to this overwhelming political instability through its history.
As a low middle-income country, Sudan is confronted with many brutal challenges, especially in health sector. Some of the challenges encountered are the poor assessment and execution of policies, lack of firm health informatics system, inadequate financial spending, centralized medical services and facilities in Khartoum and urban cities, and insufficient training for postgraduate doctors. To add more to the burden on medical care is the deficiency of preventive medicine application, poor referral system, problematic diaspora of physicians, lack of communities’ awareness leading to the fixed stigma and spiritual misconceptions of diseases that are causative of delayed medical seeking behaviors and use of folk medicine. These difficulties robustly affect the quality of health care and specifically the management of chronic diseases as epilepsy.
Neurology practice in Sudan is affected by the weakened health care system, Adult and child neurology is confronted by extreme challenges affecting people with neurological diseases. Up to the year 2005 there were only three practicing neurologists that were delivering medical care for an unconceivable ratio of one neurologist to 12 million people [4]. In addition to the enlarging population, this ratio could be attributed to lack of neurology training programs for postgraduate doctors which has begun in the past 10 years, in addition the shortage of neurology clinics in Sudan as today there are 3 tertiary neurology centers that provide adult neurology services, all of which are located in the capital which only aggravates the problem of the ability to seek neurology consultations and follow-ups especially for patients living outside Khartoum. Other major setbacks are the shortage of neuro-physiologists, neuro-imaging facilities and neuro-radiologists and the desperate need for neurology nursing and rehabilitation centers.
There is a lot of stigma and misconceptions that befall (PWE) in Sudan, where epilepsy is perceived as demonic possession, Satanic rituals, spells and witchcraft [3], some cultures have superstitions similar to that of Saudi tribes where they consider (PWE) as a presentiment of evil, a manifestation of envy and “Evil Eye” [5], while in some cultures (PWE) are considered a grace and bringers of god-bliss to their families [6]. However, others believe PWE are infectious, mentally ill, impotent and should neither get married nor have a job. Some people consider epilepsy an incurable disease, while others think the condition will pass on its’ own so they completely dismiss the therapeutic process as a futile endeavor. Some religious followers would resort to special forms of prayer involving rigorous movements to help alleviate the condition. Such beliefs direct people toward seeking traditional methods and healers, who antagonize demons, introduce herbs, ointments, cautery and prayers as standalone treatment for epilepsy.
A cross sectional study done in Sudan to evaluate the impact of spiritual and traditional believes of care givers on the management of children with epilepsy, it established that 80% of them were educated, one third of study population attributed epilepsy to supernatural causes. More than two thirds acknowledged use of both traditional and spiritual medicine, more than half used different religious methods to treat epilepsy. Almost half of participants believed that religious and or traditional treatment were truly effective in the management of epilepsy, and one third used herbs in the treatment of epilepsy [7].
In Sudan the number of centers where appropriate investigation tools has increased in the recent years, more cities are constructing new centers for neurology (like Madani neurology center, Aljazeera State), but it’s in no way comparable to the increase in patients and the services that need to be provided [8].
Despite the increase in number of medical faculties and doctors, the number of physicians with specialty training in neurology remains lacking. The overall condition of freedom and civil rights in the country along with the increased costs of living, which are all factors contributing to the mass immigration of doctors and other healthcare professionals to seek a respected income that enables them to live a decent life. It is worth mentioning that some doctors in Sudan live off salaries ranging anything from the equivalent of 15 to 300 dollars per month.
Currently, there are more than 25 licensed AEDs in clinical practice in the developed world, compared to few registered AEDS in Sudan, most of which are old generation AEDs, although older generation medications are still effective even in comparison to newer generation AEDs, the newer generation have less side effects and are more tolerable [9]. Tolerability and adverse effects are a major influence on compliance, and discontinuation of therapy, therefore increasing morbidity and mortality in people with epilepsy.
The use of AEDs is influenced by the pre-existing belief system that pushes people towards traditional herbals and local healers [3], with some believing the medication is useless while others consider s it to be undermining of the more trusted traditional methods. However, among those who would have access to proper medical help, and those who appreciate the need for medication, other factors further affect the treatment gap and challenge adherence to medication. Patients who are seizure free for a long duration or those taking more than one medication may fail to adhere to therapy or omit doses.
Descriptive analysis of cost-benefit for some patients indicates that their concern about the high price of the medication greatly outweighs the need for the drug, and would as a result seek free samples provided by charity organizations, while some patients fail to obtain the drug [10]. Antiepileptic drugs represent a tremendous economic burden on families of patients with epilepsy. The yearly cost of AEDs alone falls not less than 276 US dollars per patient per year, while visitations and consultations along with investigation could reach 51 dollars. Other indirect costs can include travel, for those who live far from the capital, reaching up to 90 dollars. Insurance rarely helps and patients find themselves forced to sell valuable assets like one’s cow or shop to cover the expenses, and many find themselves in debt. All of these factors need to be accounted for by the patient and caregivers and affect adherence negatively [11].
Access to AEDs like other medications in Sudan was subject to variations related to inflation and other complex geopolitical factors, resulting in fluctuating prices in the period from 2009 to 2013 (6 times change in pricing). And while the general market dynamics in the country were somewhat fluctuant, the general indicators of regional macroeconomics have been declining steadily (e.g. GDP in dollars) following factors like change in market policies, conflicts in the south leading to loss of big fractions of the country’s’ resources, up to the more recent financial crisis in the country in the period 2018-2020, where cash was virtually inaccessible to the public, making all medications into a luxury, and culminating in an event of pharmaceutical scarcity of drugs, despite the government’s best efforts to mitigate the impact of the economic situation [12]. Some policies had a relatively positive effect, like price liberalization privatization of the sector. And while reports and studies are yet to fully estimate the on-going catastrophe, the global status of lock-down and quarantine due to the COVID-19 pandemic certainly made it more challenging to get access to medical care or self-management for (PWE) in such a collapsing healthcare system [13].
Stigma is the social outspoken or perceived labeling of an individual or a group of people according to true or presumed different characteristics attributed to specific health related and non-health related conditions, rendering these individuals incapable of leading equal lives to their peers in society [14, 15].
Components of stigma include behavioral, emotional and cognitive elements that are portrayed in patients responses or attitudes and their interaction with society [16]. The burden of stigma unfolds in both active and passive manners, those who discriminate and those facing discrimination can inflict stigma after being subjected to it. This gives rise to the different entities of stigma and its effects on different life attributes of stigmatized individuals in society [15].
The manifestations and impact of stigma in the attitude form further branches it into perceived, anticipated, and internalized stigmas, while the social form of stigma includes the enacted or experienced stigma. Perceived stigma describes one’s thoughts or self-image perceived through the eyes of those surrounding one’s life regarding an acknowledged distinguishing characteristic [16]. Anticipated stigma refers to a presumed inappropriate response in the form of an act of discrimination or labeling in a social setting to one’s condition by others. Internalized stigma denotes self-inflicted discrediting and undermining due to awareness and acknowledgment of one’s difference. Experienced or felt stigma refers to consequences of an act of labeling or discrimination that was made intentionally to point out a stigmatizing characteristic [14, 16, 17].
Stigma is a major social determinant of health, attributing to disease morbidity, mortality and to the successfulness of healthcare services [18].
Elements that articulate the complex process of health conditions related stigma include illness nature, it’s course, and characteristics that represent origins of stigma; population related elements; treatment modalities and healthcare providers sought for consultation; reactions as well as coping mechanisms of stigmatized individuals to social acts of discrimination that may take a toll on their identity, social life, and economic thriving [17, 19, 20].
What is not so clearly defined however, is the relationship between stigma and healthcare outcomes, attributing to stigma being an entity that while having similar grounds in most health related conditions, its effects can be as illness specific as exclusive features of that illness, often referred to as the hidden burden of an illness, and this is an area that is deficient in research data [21, 22].
Health related stigma, can be visualized more clearly in communities where compensation of one’s health condition related disability is lacking. These compensations aim towards minimizing the gap between individuals with disabling health conditions and their peers in community. Communities where efforts to minimize this gap are lacking are mostly those of low-income economical index [20].
Stigma adversely affects individual health outcomes as well as related life chances, including educational opportunities, employment, housing, and social relationships. It has also been shown to negatively affect help- and treatment-seeking behaviors, compromising the ability to treat and prevent stigmatized health conditions. Masking of research on illness specific stigma under the generalization of its nature has limited the ability to understand the overall impact of stigma on individual wellbeing and the overall disease burden, restricting the ability to develop interventions addressing stigma, and this masking is amplified especially in low-income countries, because of the lacking resources available to healthcare research and services in general [20, 23].
Stigma affects caregivers of individuals being stigmatized, be it their families, relatives or close companions. Caregivers of patients in low-income countries suffer a heavier burden due to lacking national health agencies support, which widens the gap between illness-limited individuals and their peers in society, further enforcing stigma as well as worsening the financial burden. All these elements associated with stigma in low income countries develop a synergistic effect, in which each element contributes to the vicious cycle of further reducing the quality of life of stigmatized individuals [23].
The weight and burden associated with epilepsy in terms of stigma manifests with variable intensities and forms across different age groups and communities [6, 24].
Developmental aspects of one’s life including physical, mental and social development, and their bases of parenting by one’s family, education and an uninterrupted social learning experience, are affected differently with various onset age groups of epilepsy. For example, having a child with epilepsy puts tremendous pressure on the family and caregivers, especially in a low-income country where taking care of an illness free child can be troublesome. This leads to stressful parenting, creating many obstacles for a child who has epilepsy to develop at a normal rate. A child with epilepsy has a higher chance of academic underachievement, which would setback building of self-esteem and eventually in conjunction with other epilepsy related elements leads to enforcement of stigma and further disability and unsuccessful treatment, in contrast to adolescence onset of epilepsy which would have a different impact on their quality of life and would manifest in different aspects like social withdrawal despite being in a functional social and economic status. Adulthood onset of epilepsy and the manifestation of stigma associated with it could be less severe than childhood and adolescence onset and would affect one’s ability to be involved in certain elements of society, but could also be devastating in certain low-income regions with plummeting education and awareness levels, for example not being able to have a spouse in a low-income community where having epilepsy is thought to be of demonic possession [1, 6, 24].
Epilepsy in Sudan accounts for 1.6 annual mortality rates and 238.7 disability adjusted life years per 100,000. It is associated with notable stigma and social burdens. Patients with epilepsy suffer a tremendous burden of social discrimination adversely affecting their quality of life [6]. These patients are subject to being denied equal chances to a dignified life following neglect, isolation and lack of national healthcare support.
As studies in Sudan regarding epilepsy are primarily focused on clinical presentation of epilepsy, no in depth illustration or correlation between stigma of epilepsy and the outcome of epilepsy healthcare have been conducted.
However, some of the magnitude of epilepsy stigma in the Sudanese population has been captured across the different age groups of patients with epilepsy in urban and rural areas.
A study conducted by Taha et al. to identify epilepsy related stigma in the Sudanese community and to find correlation between penetrance of the type of stigma on patients through stigma degree scoring, have detected that approximately 16% of both men and women with epilepsy suffer from highly precipitated felt stigma. 12.5% of remaining patients of epilepsy who did not suffer from felt stigma have noted the common belief in their communities of the contagious nature of epilepsy while 56.2% declared their communities believed epilepsy was of demonic possession, 13% mentioned people were afraid from them when they were having seizures in public and hence they do not help them. The Sudanese community surrounding patients with epilepsy also showed poor respect to patients’ privacy evident with 77.4% of patients stating that despite not disclosing their condition, it was publicly known. Where expected least, Sudanese communities showed an alarming response to children with epilepsy from their teachers and mentors, as 22% of patients at primary school age mentioned that their teachers treated them badly. Two out of three patients with epilepsy were found to have either courtesy or coaching stigma, which represent enacted stigma of parents and guardians of patients with epilepsy, and this translates into a boosting effect for all forms of stigma being enforced in epileptic patients having their caregivers constantly reminding them of their condition. Patients who stated that their disease hindered their progress in life and those who expressed frustration and stress were found to be more than those who could cope with their condition, and this was significantly associated with a high seizure frequency. This shows that poor control of seizures inevitably diminishes the ability of patients to conceal their condition, leading to more discrimination and exacerbation of stigma [1].
An important implication of living in a resource-limited country is deficiencies that could be noted across all social services especially healthcare services. Muwada Bashir et al. portrayed a brilliant scope in their study of detecting the quality of life of Sudanese patients with epilepsy under the burden of inequalities of healthcare services, which showed that stigmatization, social discrimination and inadequate health services are major problems that Sudanese patients with epilepsy and their families confront in their daily life. The study concluded that stigma among other factors associated with epilepsy is worsening the burden on both patients and caregivers by crippling their healthcare services accessibility and by increasing efforts of coping with the disease in a society with a culture that is shaped by a low economic status [6].
Children constitute the main domain of people with epilepsy; this subpopulation faces many challenges. These challenges begin with the different etiologies of epilepsy in Sudan and Africa, of these etiologies central nervous system infectious agents (malaria, onchocerciasis), and perinatal insults constitute the main causes of epilepsy. Such causes could explain why the majority of people with epilepsy are in Africa. In addition, these causes along with other factors contribute to the poor outcome of epilepsy in the developing world.
Children with epilepsy have comorbidities including autism, intellectual disability that could be caused by perinatal insults and cerebral palsy; they are also more vulnerable to physical and sexual abuse. Studies from Sudan demonstrated that 10% of children with epilepsy have associated attention deficit hyperactivity disorder (ADHD) [25], one third had learning disabilities, and 10% had motor disabilities [26], theses comorbidities represent the difficulty in the management of these children, as a multidisciplinary approach is required in management, which is usually unavailable in Sudan and the developing world.
Since the 1950s, children with neurological disorders were seen in adult neurology clinics, as pediatric neurology training program in Sudan has recently been initiated, with a few pediatric neurologists available.
Currently there is one pediatric neurology tertiary center and four specialized child neurology clinics in Sudan, 3 of them are located in the capital, these 4 clinics serve the whole of Sudan, as well as referred patients from neighboring countries including: Chad, Eretria, and South Sudan where facilities for neurological investigations are limited. The shortage of pediatric neurologists and pediatric neurology centers and their location mainly in the capital, along with the high cost of transportation to the center, long waiting lists till evaluation by a specialist, further complicate the management of children with epilepsy [8].
It is important to review epilepsy status in school settings where children spend most of their time. Schools in Sudan rarely have dedicated clinics to accommodate children’s health needs, and while school teachers should act as caregivers, most of them are usually ill-informed or lacking appropriate knowledge about epilepsy, and none of them have had any sort of training to help in case of a seizure, so a considerable proportion does not know what to do when a child develops a seizure [27, 28]. Many teachers fall as victims of the communities’ misconceptions and could even play a passive role in the stigma, contributing to the child’s anxiety. Many had no idea about possible causes of epilepsy and guessed that parents would not sign up their children with epilepsy to school due to suspected mental sub-normality, stigma, or fear of unattended falls or attacks. On the other hand, figures demonstrated a significant amount of children ditch school altogether because of the illness. Other students do not mind having a classmate with epilepsy at school but they share their teachers’ beliefs and misconceptions, and would sometimes, as a result, engage in bullying and discriminatory behaviors against them. The condition is barely touched in school curriculums and students do not undergo any sort of training to help them act properly around their peers who have epilepsy.
Globally, 50% of women and girls with epilepsy are in the reproductive age range [29]. Epilepsy in the developing countries has a slight male predominance; this is likely due to underreporting of epilepsy in women due to negative attitudes and stigma facing them, that include difficulties in getting married, increased divorce rates, having children or even being abandoned by their families because of their illness, and harder chances of being employed. This underreporting of epilepsy in women leads to deficits in health care seeking behavior, hence contributing to the epilepsy treatment gap in women.
Apart from the aforementioned social difficulties, women with epilepsy are challenged with many issues that include the effect of epilepsy and AEDs on their sexual function, contraception, pregnancy, fetal abnormalities, childbirth, and breastfeeding [30, 31, 32, 33].
Due to the shortage of neurologists in Sudan, the majority of women with epilepsy are managed and counseled by non-specialized doctors. A study conducted in Sudan to assess doctors’ knowledge of women issues and epilepsy using standardized knowledge of women issues and epilepsy (KOWIE II) questionnaire concluded that the majority of Sudanese doctors’ knowledge was unsatisfactory. They were unaware of sexual dysfunction among women with epilepsy, that women with epilepsy should continue taking their AEDs when they are pregnant, and that women can safely breastfeed while taking AEDs [34].
Sudan has been a victim of war, poverty, substandard infrastructure, and a failing healthcare system. These factors along with epilepsy stigma, misconceptions and false believes represent major challenges in epilepsy management in Sudan.
All these challenges must be approached systematically to ensure the best management for patients with epilepsy. Such approaches include the need for a mass movement against epilepsy headed by individuals experienced in the field, and fundamental governmental partnership and aid to provide organizational efforts and funding for instituting and decentralizing neurology facilities outside Khartoum, and ensuring the availability and affordability of investigations and medications especially the new generation AEDs. Epidemiologic studies are needed to outline the treatment gap of epilepsy and guide nationwide strategies and efforts to increase the awareness of communities about epilepsy are needed especially in the rural areas to fight disease stigma, Special groups need further attention such as making efforts for prevention of infections leading to epilepsy in children, the involvement of other healthcare providers such as social workers, speech and language therapists, nutritionists, and special teachers in the management of children with epilepsy can never be overemphasized. Lastly, telemedicine should be implemented in the management of epilepsy in Sudan.
The authors declare no conflict of interest.
Ismat Babiker wrote the following sections: children with epilepsy, women with epilepsy, co-wrote AEDs in Sudan, and contributed in chapter editing.
Awab Saad wrote Sudan: background and population, healthcare system in Sudan, Neurology in Sudan, co-wrote epilepsy misconceptions in Sudan, and contributed in chapter editing.
Basil Ibrahim wrote stigma, health related stigma, manifestation of stigma in high vs. low-income countries, stigma in low-income countries and in Sudan, and contributed in chapter editing.
Mohamed Abdelsadig wrote the collapse of the healthcare system in Sudan, epilepsy in schools, scarce personnel and trained physicians, co-wrote AEDs in Sudan, epilepsy misconceptions in Sudan, and contributed in chapter editing.
Supporting women in scientific research and encouraging more women to pursue careers in STEM fields has been an issue on the global agenda for many years. But there is still much to be done. And IntechOpen wants to help.
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