**2.2 Filaggrin and altered skin barrier function**

AD is characterized by eczematous skin lesion, dry skin, pruritus, increased TEWL, and enhanced percutaneous penetration of both lipophilic and hydrophilic compounds (Jakasa, et al., 2011, Wollenberg &Bieber, 2000). The skin barrier defect is one of the primary events that initiate disease pathogenesis, allowing the entrance of numerous antigens into the epidermis in patients with AD (Onoue, et al., 2009, Osawa, et al., 2011). The *FLG* mutation carriers have demonstrated elevated TEWL (Jungersted, et al., 2010, Kezic, et al., 2008), basal erythema, skin hydration, increased skin pH (Jungersted, et al., 2010, Nemoto-Hasebe, et al., 2009), SC thickness (Nemoto-Hasebe, et al., 2009), impaired SC integrity upon repeated tape stripping (Angelova-Fischer, et al., 2011), and increased diffusivity of PEG 370 (Jakasa, et al., 2011) compared to healthy donors. Nevertheless, these alterations found in *FLG* mutation carriers are not consistently correlated with AD since AD patients without *FLG* mutation might also share some similar features. (Hubiche, et al., 2007, Jakasa, et al., 2011, Jungersted, et al., 2010, Kezic, et al., 2008). It is, therefore, suggested that other factors besides *FLG* lossof-function mutations modulate skin barrier integrity, especially in AD.

Since the skin barrier is related to intercellular lipid bilayers of the SC, it might be interesting to examine the composition and the organization of intercellular lipids of the SC in AD patients in relation to *FLG* genotype and disease severity (Jakasa, et al., 2011). Carriers of *FLG* mutations showed significantly reduced levels of NMF in the SC (Kezic, et al., 2008). Similar lipid composition of *FLG* mutation carriers and individuals with normal filaggrin was observed (Angelova-Fischer, et al., 2011, Jungersted, et al., 2010), but a lower cermide/cholesterol ratio was detected in the former group (Angelova-Fischer, et al., 2011). Filaggrins proteolytically degraded into a pool of free amino acids including histidine and glutamine which are further converted to, respectively, urocanic acid (UCA) and 2-

Flaky Tail Mouse as a Novel Animal Model of Atopic Dermatitis:

**3.1 Origin of flaky tail mice**

Presland, et al., 2000).

copies of the filaggrin repeat (Fallon, et al., 2009).

two to four weeks (Jarret A, 1957, Searle A.G., 1957).

et al., 2010, Presland, et al., 2000) (Fig.1).

Possible Roles of Filaggrin in the Development of Atopic Dermatitis 7

same result could be seen among HLA-DRB1\*1501 (a HLA class II complex which is immunodominant in individuals with AD (Ardern-Jones, et al., 2007)) positive adult individuals with moderate-to-severe AD and *FLG* mutations (McPherson, et al., 2010).

The above findings indicate the involvement of filaggrin in the development of AD. Therefore, the impact of filaggrin deficiency on cutaneous biological functions *in vivo* should

Flaky tail mice (*Flgft*), first introduced in 1958, are spontaneously mutated mice with smaller ears, tail constrictions, and a flaking tail skin appearance (Lane, 1972). *Flgft* mice were outcrossed onto B6 mice at Jackson Laboratory (Bar Harbor, ME, USA) (Lane, 1972, Presland, et al., 2000) (Note: Although this strain was crossed with B6, it is not a B6 congenic strain but rather a hybrid stock that is probably semi-inbred). Homozygous *Flgft* mice have dry, flaky skin which expresses reduced amounts of profilaggrin mRNA and abnormal profilaggrin protein that is not processed to filaggrin monomers (Fallon, et al., 2009,

Recently, it has been revealed that the gene responsible for the characteristic phenotype of *Flgft* mice is a single nucleotide deletion at position 5303 in exon 3 (5303delA) of the profilaggrin gene, resulting in a frameshift mutation and premature truncation of the predicted protein product. The copy number of the filaggrin repeat contained within this gene varies depending on the background strain. This mutant occurs in an allele with 16

*Flgft* mouse carries double gene mutation, *Flg* and matted (*ma*) in which the locations of the mutated genes are within close linkage to one another (Lane, 1972). The *ma* gene characteristic reported by Searle & Spearman (1957) causes the body-hair of affected mice to be brittle and inflexible, which results in longitudinal splitting and breaking due to friction against the cage and other objects. This mutation is a fully penetrant recessive house-mouse mutant which belongs to the "naked" category (i.e., a house-mouse with baldness resulting from the breaking of hairs or from hereditary hairlessness). This mutation can be identified morphologically by (1) erection of hairs, (2) matting of hair in clumps, (3) a tendency towards baldness, (4) a change from black- to brown-colored melanin in old hairs. The age at which this mutant is first identified based on external appearance varies from between

Recognition of the features of this mouse is more evident between 5 and 14 days of age when constricted, flaking tail skin and thickened short pinna of the ears are observed. In addition, *Flgft* mice are often smaller than their normal siblings at this age. Routine histological sections stained with hematoxylin and eosin showed that the stratum granulosum in *Flgft* mice at 1, 2, 4, and 8 days of age does not contain as many granular layers as that of non-*Flgft* mice (Lane, 1972). Mice of the *Flgft* genotype express an abnormal profilaggrin polypeptide that does not form normal keratohyalin F-granules and is not proteolytically processed to filaggrin. Therefore, filaggrin is absent from the cornified layers in the epidermis of the *Flgft* mouse (Fallon, et al., 2009, Presland, et al., 2000, Scharschmidt, et al., 2009). Consistently, we and others have described that *Flgft* mice express a truncated and smaller profilaggrin protein that is not processed to filaggrin (Fallon, et al., 2009, Moniaga,

**3. Flaky tail mouse as a novel animal model of atopic dermatitis**

be analyzed in detail. To address this issue, animal models are of great value.

pyrrolidone-5-carboxylic acid (PCA). The concentrations of UCA and PCA in SC in the carriers of *FLG* mutations were significantly lower than those in healthy donors (Kezic, et al., 2009). Therefore, filaggrin deficiency is sufficient to impair epidermal barrier formation.

An *in vitro* experiment using filaggrin knocked down human organotypic skin cultures showed enhanced penetration of hydrophilic dye Lucifer yellow, smaller lamellar bodies, and deficiency of their typical lamellae without altered lipid composition (Mildner, et al., 2010). In addition, UCA, one of the filaggrin-derived free amino acids and as an important UV absorbent within SC, was decreased following filaggrin knocked down, leading to increased sensitivity to UVB-induced keratinocyte (KC) damage (Mildner, et al., 2010).

#### **2.3 Filaggrin and altered immunobiology**

The SC serves as a biosensor of the external environment and a link between innate and adaptive immune systems (Vroling, et al., 2008). The critical association between the abnormal barrier in AD and Th2 polarization may in part be explained by the production of the cytokine, thymic stromal lymphopoietin (TSLP) (Ebner, et al., 2007).

TSLP has been shown to activate dendritic cells to drive Th2 polarization, through upregulation of the co-stimulatory molecules CD40, CD80, and OX40L, triggering the differentiation of allergen-specific naïve CD4+ T cells to Th2 cells that produce IL-4, IL-5, and IL-13 (Ebner, et al., 2007, Soumelis, et al., 2002).

Patients with Netherton syndrome (NS), a severe ichthyosis in which affected individuals experience a significant predisposition for AD, have elevated levels of TSLP in their skin. Upregulated kallikrein (KLK) 5 in the skin of NS patients directly activates proteinaseactivated receptor 2 (PAR-2) and induces nuclear factor kappaB-mediated overexpression of TSLP, intercellular adhesion molecule 1, TNF-α, and IL-8. This phenomenon occurs independently of the environment, adaptive immune system and underlying epithelial barrier defect (Briot, et al., 2009, Briot, et al., 2010). *In vitro* study using human keratinocyte cell line HaCaT cells and reconstituted human epidermal layers transfected with filaggrin siRNA showed increased production of TSLP via toll-like receptor (TLR) 3 stimulation (Lee, et al., 2011). These findings suggest that reduced filaggrin levels may influence innate immune response via TLR stimuli and elevate TSLP, leading to AD-like skin lesions.

AD is one of the emerging diseases in which epidermal dysfunction increases allergen and microbial penetration in the skin, with the consequent development of adaptive Th2 immune responses (Kondo, et al., 1998) within regional lymphoid tissue. The resultant Th2 cells may then home back to the skin or lungs, where they recognize allergen in the skin (McPherson, et al., 2010), which leads to local Th2 inflammation, reduced antimicrobial peptide expression (Nomura, et al., 2003), and filaggrin downregulation (Howell, et al., 2007). Indeed, the induction of circulating allergen-specific CD4+ T cells may be an important prerequisite underlying the pathogenesis of the atopic march (O'Regan, et al., 2009). Among moderate-to-severe AD patients, the *FLG* mutation carriers showed a greater number of house dust mite Der p1-specific IL-4 producing CD4+ T cells, suggesting that filaggrin mutations predispose to the development of allergen-specific CD4+ Th2 cells. The same result could be seen among HLA-DRB1\*1501 (a HLA class II complex which is immunodominant in individuals with AD (Ardern-Jones, et al., 2007)) positive adult individuals with moderate-to-severe AD and *FLG* mutations (McPherson, et al., 2010).


**Synthesis, Optical and Electrical Characterization of Amino-alcohol Based Sol-gel Hybrid Materials**

**Bárbara R. Gomes, Rita B. Figueira \* , Susana P. G. Costa , M. Manuela M. Raposo and Carlos J. R. Silva** †

Centro de Química, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal<EMAIL_ADDRESS>(B.R.G<EMAIL_ADDRESS>(S.P.G.C<EMAIL_ADDRESS>(M.M.M.R.)

Received: 22 October 2020; Accepted: 10 November 2020; Published: 12 November 2020

**Abstract:** This manuscript describes the synthesis and characterization of five new organic–inorganic hybrid (OIH) sol-gel materials that were obtained from a functionalized siloxane 3-glycidoxypropyltrimethoxysilane (GPTMS) by the reaction with the new Jeffamine®, namely three different diamines, i.e., EDR-148, RFD-270, and THF-170, a secondary diamine, i.e., SD-2001, and a triamine, i.e., T-403. The OIH sol-gel materials were characterized by UV-visible absorption spectrophotometry, steady-state photoluminescence spectroscopy, and electrochemical impedance spectroscopy. The reported OIH sol-gel materials showed that, with the exception of the samples prepared with Jeffamine® SD-2001, the transmittance values ranged between 61% and 79%. Regarding the capacitance data, the values reported changed between 0.008 and 0.013 nF cm−2 . Due to their optical and electrical properties these new OIH materials show promising properties for applications as support films in an optical sensor area such as fiber sensor devices. Studies to assess the chemical stability of the OIH materials in contact with cement pastes after 7, 14, and 28 days were also performed. The samples prepared with THF–170 and GPTMS, when compared to the samples prepared with RFD-270 and T-403, exhibited improved behavior in the cement paste (alkaline environment), showing promising properties for application as support film in optical fiber sensors in the civil engineering field.

**Keywords:** hybrids; sol-gel; materials; amino-alcohol


**2.1 Dry spinning**

*Science, Technology and Advanced Application of Supercapacitors*

used [4–6].

[17–22].

mentioned properties.

ion diffusion paths [35–37].

tive electrodes.

far higher than that of EDLCs.

operate for extended periods of time, often millions of cycles without losing their energy storage capacities giving them an edge over batteries in how long they can be

Supercapacitors have two main classifications that are based on their charge storage mechanism and the type of electrode materials [4]. The first one, electric double layer capacitor (EDLC), stores charges electrostatically on the electrodeelectrolyte interfaces of the high surface area carbon materials. This process involves physical adsorption of ions at the electrode and electrolyte interface [2]. The second one, pseudocapacitor, on the other hand, stores charges within the electrodes in response to fast surface and near-surface redox reactions [5, 7]. The electrodes are derived from transition metal oxides and conducting polymers. Due to these redox reactions, pseudocapacitors have been reported with energy densities

With the emergence of flexible electronics such as foldable displays [8], soft photo-detectors [9] and bendable field effect transistors [10], flexible supercapacitors have become more popular than ever. They have been found to be suitable in powering portable and flexible electronic devices, and several have been fabricated with lightweight, flexible and possessing high power and energy densities [11–14]. Planar format supercapacitors have been found to have larger volumes and structural limitations which impede their use in lighter, smaller and omnidirectional flexible electronic devices [15, 16]. To solve these problems, lightweight and high energy density fiber-shaped supercapacitors have been explored and fabricated

Fiber electrodes for supercapacitors have been made from active materials with

Polyaniline (PANI) is probably the most widely studied of the conductive polymers because of its high electronic conductivity, redox and ion exchange properties, excellent environmental stability and ease of preparation [32–34]. It has, therefore, been extensively explored in energy storage devices fabricated with pseudocapaci-

Bulk PANI, however, due to its low accessible surface area is not ideal for energy storage device electrodes. The workaround to this has been to fabricate nanostructured PANI materials. These structures have typically been made using a carbon template thereby producing materials with a large area to volume ratio and shorter

In this chapter, we report our high energy density fiber supercapacitors based on CNT-PANI fiber composites. A chemical oxidation polymerization technique is employed to deposit PANI on the surface of the CNT fibers. This composite material gives superior performance as supercapacitor electrodes due to the fast redox reactions between the PANI and the electrolytes used. To create our CNT fibers, we employ a technique that involves dry spinning of multi-walled carbon nanotube (MWCNT) fibers from vertically aligned MWCNT arrays grown by chemical vapor deposition (CVD) as described in a previous publication by our research group [38]. This technique is used to spin continuous fiber at industrial rates from MWCNT arrays of 3 cm width and 4.25 cm length, resulting in fibers with diameters of approximately 55 μm and up to 40 m in length. Next, these fibers underwent atmospheric pressure oxygen plasma functionalization to create oxygen plasma functionalized CNT (OPFCNT) fibers as the base structure for the PANI deposition.

nanostructures, such as CNTs [23–25], graphene [17, 26, 27] and metal oxides [28–30]. However, the most widely studied ones have been CNT fiber electrodes and their composites. This is attributed to CNT's inherent flexibility, high surface area and high electrical conductivity [31]. In their fiber formats, they are highly aligned and have excellent mechanical durability while maintaining all their afore-

**40**

CNT fibers are dry spun from vertically aligned CNT arrays. In our work, thin films of Fe and Co were sputtered on a silicon wafer, overlaid with approximately 5 nm Al2O3 as a buffer layer, by means of physical vapor deposition (PVD). The created structure serves as a catalyst for the growth of aligned CNT arrays on the silicon wafer. This surface-treated substrate was then diced up into required pieces and then exposed to a CVD environment in a FirstNano ET3000 reactor. The resulting CNT array was drawable and spinnable and, by means of twisting and pulling. A homemade setup was used to fabricate highly aligned fibers [38], as shown in **Figure 1**. These pristine CNT fibers, when used to form EDLCs, produce quite low energy densities necessitating the deposition of PANI on them to increase the energy density.

## **2.2 Oxygen plasma functionalization of fibers**

After the fibers were spun, they underwent an atmospheric pressure oxygen plasma functionalization process to improve the wettability of the fibers. This is necessary since carbon-based materials are naturally hydrophobic and need improved wettability to increase the deposition of PANI on the surface of the fiber during the oxidative polymerization process [39–42]. In previous publications [43, 44], carbon-based materials were treated with acids to functionalize them and thereby improve the wettability before polymerization. These involve wet chemistry and as such mostly require multi-step reactions and involve strong chemicals, which affect the bulk properties of the CNT structures. The plasma functionalization process employed in this work is continuous, effective and can be used industrially for extensive lengths of fibers.

Oxygen plasma functionalization was generated by systematically pulling the CNT fiber through a plasma head with a chamber for tubular structures (Surfx Atomflo 400 system). The set-up is shown below in **Figure 2**. The pristine CNT fiber was threaded through the plasma head and affixed to the collector bobbin with double-sided tape. The fiber was pulled through the plasma head at a speed of 0.206 cm/s using the collector bobbin on the motor. This processing led to the functionalization of the fiber with the following plasma parameters: 60 W power, 0.1 L/min oxygen and 15 L/min helium. These parameters were chosen since they ensured the fibers to be functionalized had minimum destruction, checked by Raman spectroscopy.

**Figure 1.** *Carbon nanotube fiber spinning process by twisting and pulling.*

**Figure 2.** *Oxygen plasma functionalization set-up for CNT fibers.*

### **2.3 Chemical oxidation polymerization**

The oxidation of aniline in an acidic aqueous medium using ammonium peroxydisulfate (APS) as an oxidant is widely used and reported in the literature [45, 46]. Emeraldine salt (green color) is the form of PANI obtained during this process [32, 47]. PANI can exist in three oxidation states: leucoemeraldine (fully reduced), emeraldine (partially oxidized) and pernigraniline (fully oxidized) [32, 45–47].

At a pH of less than 2.5, the oxidative polymerization of aniline is a chain reaction [48]. The growth of the chains proceeds by the addition of the monomeric aniline molecules to the active chain ends. The chain growth is terminated after at least one of the reactants in the polymerization runs out. If there is an excess of the APS (oxidant), the resulting polymer remains in the pernigraniline form [49], especially at molar ratios of APS to aniline of over 1.5. If the rate of APS to aniline is equal to 1.25 [50] or aniline is in excess, pernigraniline is reduced to emeraldine at the end of the reaction while aniline is oxidized at the same time to emeraldine [48, 51]. We, therefore, ensured in all our tests that we had excess aniline to promote emeraldine growth, the most thermally and environmentally stable form of PANI [52–54].

The oxygen plasma functionalized CNT (OPFCNT) fibers were cut into 7.5 cm portions and affixed to copper tapes with fast drying silver paint (TedPella Inc.). The copper tapes served as the leads used to connect the devices for electrochemical testing. These electrodes were then placed into 10 ml beakers and put into an ice bath. Aniline monomer dissolved in 1 mol/L HCl and the ammonium persulphate (APS) solution also dissolved in 1 mol/L HCl were then put in the various beakers with fibers at different ratios of aniline to APS. The amount of PANI formed on the fibers was controlled by the ratio of aniline to APS used as well as the time the solution was allowed to polymerize. The fibers were taken out after the polymerization time and rinsed in a beaker with deionized water to wash off the excess PANI.

#### **2.4 Electrode and device fabrication**

Fiber supercapacitors were created using poly (vinyl alcohol) and sulfuric acid (PVA-H2SO4), as well as polyvinylidene fluoride-co-hexafluoropropylene and 1-ethyl-3-ethylimidazolium (PVDF-EMIMBF4) gel electrolytes. The PVA-H2SO4 was made with 10 ml DI water, 2 ml H2SO4 and 1 g PVA. The PVDF-EMIMBF4 gel electrolyte was prepared with 15 ml acetone, 1.5 g PVDF, and 3 ml EMIMBF4. The PVA-H2SO4 was operated at a 1 V window, while the PVDF-EMIMBF4 was operated at a 3.2 V window. The larger voltage window the PVDF-EMIMBF4 allowed enabled

**43**

*Fiber Supercapacitors Based on Carbon Nanotube-PANI Composites*

us to reach larger energy densities. Devices were made from these fibers by coating them with the gel electrolyte (PVA-H2SO4 or PVDF-EMIMBF4). The fibers were then placed parallel to each other on a weighing sheet, with more electrolyte and

Electrochemical measurements were carried out with an electrochemical workstation (Gamry, Interface 1000) at room temperature. The electrochemical characteristics of the electrodes and devices were evaluated by cyclic voltammetry at various scan rates, galvanostatic charge-discharge tests, and electrochemical

amplitude of 10 mV at the open circuit potential. In a three-electrode configuration test, Ag/AgCl was used as the reference electrode, platinum served as the counter

The capacitance (*C*) of the electrodes and fiber supercapacitors was calculated from the galvanostatic discharge curves at different current densities by using the equation: *C=IΔt/ΔV.* The gravimetric capacitance (*Cm*) and areal capacitance (*CA*) were calculated by the following formula: *Cm = C/m* and *CA = C/A,* respectively. The gravimetric energy density (*Em*) and power density (*Pm*) were calculated by the

expressions: *Em = 1/2(Cm(ΔV)2)/3.6* and *Pm = 3600Em/t*.*Pm* = 3600\_\_\_

and *A* refer to the mass and volume of the device, respectively [40, 55].

copy (Renishaw inVia, 514 nm Ar-ion laser with a laser spot of ~1μm2

2 ∗ *Cv* (*V*)2 \_\_\_\_\_\_\_

taken for each sample on at least two different locations.

0.776 to 1.195, signifying the destruction of the carbon sp2

energy density (*EA*) and power density (*PA*) were calculated by the expressions:

the discharge current, *t* is the discharge time, *ΔV* is the operating voltage window, *m*

Scanning electron microscopy (SEM) (FEI XL30, 5 kV) and Raman spectros-

characterize the CNT-PANI. The masses of the fibers were taken on a Sartorius SE2 ultra-microbalance. X-ray photoelectron spectroscopy (XPS) data were obtained using a VG Thermo-Scientific MultiLab 3000 ultra-high vacuum surface analysis

energy. The high-resolution scans for carbon and low-resolution survey scans were

The plasma functionalization of the fiber was confirmed by Raman data **Figure 3a** and XPS data **Figure 3b–d**. From the Raman spectra in **Figure 3a**, we observe an increment in the ratio of intensities between the D and G peak, from

functionalization. In **Figure 3b**, there is a documented increase in the atomic weight percent of oxygen from 9.1% in the pristine state to 28.17% for the oxygen plasma functionalized thread. **Figure 3c** and **d** is deconvoluted high-resolution C1s and O1s peaks from the XPS data, showing the various oxygen functional groups found on the surface of the fiber which is in close agreement with data

PANI-CNT composite fibers were created from four ratios of aniline to APS

(1:1, 2:1, 5:1 and 10:1). The OPFCNT fibers were placed with the chemicals as they polymerized for an hour. From our electrochemical half-cell tests, we observed that a 2:1 aniline to APS ratio gave the best specific capacitance, as seen in **Figure 4a**. Further testing of OPFCNT fibers with varying durations (10 minutes to 6 hours) of polymerization revealed that the composite fibers that

to 10−1

3.6 and *PA = 3600EA***/t***, PV* = 3600\_\_\_

Torr base pressure using an Al Kα source of 1486.6 eV excitation

Hz using sinusoidal voltage

*Em*

*EV*

bonds during plasma

*t* The areal

*t* where *I* is

) were used to

*DOI: http://dx.doi.org/10.5772/intechopen.80487*

**2.5 Electrode and device characterization**

impedance spectroscopy measurements from 106

electrode and the experiments were run in 1 M Na2SO4.

sealed with Kapton tape.

*EA = 1/2(CA(ΔV)2)/3.6 EV* = \_1

system, with ~10−9

**3. Results and discussion**

reported in the literature [39, 56].

us to reach larger energy densities. Devices were made from these fibers by coating them with the gel electrolyte (PVA-H2SO4 or PVDF-EMIMBF4). The fibers were then placed parallel to each other on a weighing sheet, with more electrolyte and sealed with Kapton tape.


**6. Quantitative research: method**

At the start of the study 46 advanced pianists (aged 18–26 years and of mixed gender) from three higher educational institutions were randomly assigned to a therapy or control group. The therapy groups received two interventions of either CH or EMDR during a two-week period between two concerts. A quantitative assessment of the state component of cognitive anxiety of all participants was obtained from the Spielberger State-Trait Anxiety Inventory (STAI-Y1) taken at baseline and prior to both performances. The findings reported here are on state anxiety; for the full findings of the author's research, including trait anxiety, somatic/physiological symptoms of anxiety and the behavioural aspects of performance anxiety see Brooker [3].

#### **6.1 Results**

At the end of data collection to establish whether anxiety levels decreased more in the intervention groups than the Control, an ANCOVA was calculated comparing levels of state anxiety across the three groups at the first and second performances (before and after treatment). There was a main effect of condition (F (2, 42) = 4.92, *p* = .012) such that participants in the two treatment conditions, CH and EMDR, showed significantly lower cognitive anxiety than the Control group at the second performance, post-intervention and that both treatment groups were significantly effective in achieving this (Helmert contrasts, .005 significance level).

This suggests that the therapies applied between the two performances significantly lowered anxiety in both the CH and EMDR groups prior to the second performance and this required two sessions only. This effect was not demonstrated in the Control group.

**Figure 1** below shows the standard error of the mean score of state anxiety at the two performances calculated from the STAI Y-1 questionnaire, where 80 represents the highest level of anxiety and 20 the lowest. It illustrates the decrease in state anxiety at the second performance across the three groups.

#### **7. The qualitative research: case study**

Qualitative information on music performance anxiety (MPA) is documented here in the form of a case study, taken from the author's private practice [47]. A personal account of performance experience is given which allows for more detail, sensitivity and insight into the understanding of MPA which could not be obtained from quantitative research alone. It gives insight into the process of therapy by

#### **Figure 1.**

*The standard error of the means of state anxiety from the STAI Y-1 questionnaire taken 15 min prior to performances 1 and 2: error bars show 95% CI of mean.*

exploring the internal thoughts, feelings and experience of the individual. This case study begins with the individual's own words (the narrative). It then continues with an abbreviated description of the progression of treatment and documents the therapeutic outcome, as well as reflecting on the suitability of the treatment administered from a research standpoint and comparisons of possible treatment effects of other treatments. It is the documentation of an individual who received one therapy of EMDR and one of CH for the treatment of MPA, (the name is anonymised).

#### **Identifying information**

Name: Rebecca.

Music performance anxiety: Voice.

Age: 41.

Occupation: Student with the Open University studying English and Music (first instrument, voice).

#### **7.1 Case history: Rebecca's narrative**

*I had always felt very nervous when performing and always seemed to have the image of my father in the background; I felt that he was judging and criticising me and that I was letting myself down. I was having singing lessons at this time and giving small informal recitals but I was never happy with the outcome; I always felt I fell short of my*

*Cognitive Hypnotherapy and EMDR: Two Effective Psychodynamic Therapies for the Rapid… DOI: http://dx.doi.org/10.5772/intechopen.101770*

*true potential. I took Grade 8 ABRSM Singing examination but failed quite badly and this felt like the ultimate disaster. Although I was confident practising on my own, I found singing lessons 'nerve racking' and at the examination I just 'fell apart'.*

*After I moved here and restarted singing lessons I was very interested in taking part in the Christmas concert as your research was close to my heart as I always get very nervous in performance. I was singing in the second half of the concert but felt nervous in the first half and this got worse, and as soon as I started singing my throat felt tense and I had saliva in my mouth which I had to keep swallowing before the long runs, my heart was beating quickly and I could feel my cheeks flushing. Because of all this I felt that I had not connected with the audience and was disappointed with my performance. I did enjoy taking part however and being part of the research into music performance anxiety and found it somewhat reassuring that others taking part were also feeling nervous, I wasn't the only one.*

#### **7.2 Case formulation**

*Therapist's summary and interpretation of Rebecca's narrative.*

Important features in this narrative indicate that the domineering personality and the physically abusive behaviour of her father, who was against her musical ambitions, have had far-reaching consequences on Rebecca's singing performances both as a teenager and as an adult. These emotions are so strong in present-day performances that she imagines the spectre of her father while she is performing. The negative criticism that she experienced from her father has affected her selfesteem and self-worth as a musician in spite of her belief that she has talent as a singer.  She is confident practising on her own where she perceives that she is not being judged; however she experiences both cognitive anxiety and distressing physiological and somatic symptoms of anxiety in a performance or examination situation where she feels she 'falls apart'.

#### **7.3 Critical analysis: therapist**

An analysis of Rebecca's narrative suggests that she is suffering from social phobia in situations where she feels threatened and under scrutiny, such as a singing examination or a live concert performance. Social phobia, rooted in social anxiety, has been summarised [84].

Rebecca experiences cognitive anxiety as well as physiological and somatic symptoms of anxiety in a performance situation. However, although she experiences destructive and crippling anxiety when performing, this phobic reaction does not occur in other areas of her life. The therapist purports that this is as a result of the criticism and physical abuse that she received from her father during her teenage years regarding her ambitions as a singer ('my father literally tried to knock it out of me') and this I believe has had a profound psychological effect on her selfesteem and confidence in her music performances.

In therapy Rebecca presented with the following negative schemas regarding her anxiety (direct quotes):

'I know I'll screw it up'

'I can't control my thoughts'

'I'm hopeless'

'I can't control my emotions'

'I can't control my body'

'I can't control my nerves'

Physiological and somatic symptoms of anxiety were:

rapid heartbeat;

shaking/trembling;

tension in throat;

an excess of saliva.

#### **7.4 Treatment plan: EMDR and CH**

#### *7.4.1 First treatment (EMDR): 30 May 2010*

Rebecca's MPA is complex as it is not related to a single incident but the systematic criticism that she received as a teenager, undermining her self-belief in the possibility of her pursuing a professional career in music as an adult. The primary aspect of the overall treatment would be targeting the trauma experienced at this time which in the opinion of the therapist is best addressed initially through EMDR by systematic desensitisation of these experiences.

In therapy the negative criticism and upsetting experiences revealed in Rebecca's narrative regarding her father were the main targets, beginning with the most upsetting incident which she rated as 9/10 on the subjective unit of disturbance scale (SUD) (10 being the highest level of anxiety). This rating indicates that significant trauma had been experienced at this time. The most painful negative emotions when recalling this incident were of fear and anger; the strongest physical sensation was tension throughout her whole body which was accompanied by heightened breathing. After fifty minutes of EMDR her rating on the SUD scale decreased to 0, indicating that the negative memories had been desensitised. The negative schemas that she had presented with at the start of treatment had now changed. Where previously she had six negative self-perceptions (four beginning with 'I can't … '; see above), post-treatment these had changed into positive perceptions of 'I can … '). She no longer thought of herself as being hopeless or that she would 'screw it up'.

Her rating on the validity of cognition scale (VOC scale was 6/7 (7 being the highest level of positivity): negative schemas cited earlier had been reprocessed. Her bodily sensations which she had experienced at the start of therapy when recalling the traumatic memories had now gone completely, her breathing had normalised and she had no tension anywhere in her body. Having targeted and desensitised the most traumatic memories first, the lesser memories of trauma regarding performance when reviewed were more difficult to hold, and no longer caused Rebecca

*Cognitive Hypnotherapy and EMDR: Two Effective Psychodynamic Therapies for the Rapid… DOI: http://dx.doi.org/10.5772/intechopen.101770*

the former anguish or physiological/somatic symptoms of anxiety. If the past has been one of negativity or trauma regarding aspects that are important to the individual, the subjective behavioural response to a similar present-day experience will be consistent with the negative affective responses of the past [56]. An adult may experience feelings of fear and being out of control, and will react emotionally and display negative behaviour accordingly.

#### *7.4.2 Second treatment (CH): 6 June 2010*

The second treatment session (1 h) was shorter than the first (90 min), the important groundwork having been accomplished in the first session. As the disparate memories had been desensitised and reprocessed in the first session CH should now be beneficial in supporting the reprocessed cognitive perceptions. It should also enhance the positivity achieved in the previous session. Rebecca had experienced hypnotherapy some years previously but it had not been particularly effective. However it was explained that it should enhance the EMDR treatment and the combination of the two therapies would strengthen treatment effects. The most important aspects of her singing performance were discussed; she wanted to feel confident, calm and in control in performance, to connect with her audience and feel eager to do more. Her key words, which were 'anchored' on her dominant wrist during hypnotherapy, were *confident*, *calm* and *in control*; these she felt were the words that would enable her to give her optimum performance. The therapy focused on enhancement of performance and included visualisation of her perfect performance.

On completion of hypnotherapy Rebecca was given the therapist's *Self-Confidence for Musicians* CD and advised to listen to this as often as possible, and especially on the day/evening prior to a performance; this would further relax her and add to her confidence. She left feeling happy and relaxed looking forward to her next singing lesson and her next performance.

#### **7.5 Rebecca's self-assessment of treatment**

*My singing in front of audiences can be adversely affected by pre-performance nerves, which seem to stem from my first solo concerts when I suffered with acute stage fright. The EMDR treatment brought my worst singing nightmares to the surface and I was initially sceptical that anything could be done to help my anxiety when performing. However I came away from the first treatment feeling unburdened as if a weight that I had been carrying around for years had lifted. It was like coming out from underneath a dark cloud. The CH treatment in the second session focused my mind on enjoying singing so that when I performed I was in control and relaxed. It appeared to reinforce everything so that I am now looking forward to performing instead of dreading it.*

*(Email message, June 2010)*

### **7.6 Therapist's assessment of treatment and reflections from a research standpoint**

This case study supports current research into anxiety which suggests that negative affect and beliefs from the past control the individual in the present; however they can be healed quickly, effectively and profoundly when past negative-rooted traumas are changed [58, 64]. The negative cognitions and previous perceptions of subjective performance that this patient held were successfully desensitised and reprocessed in two treatments.

#### *Complementary Therapies*

There can be no doubt that the treatment was effective; however reflecting on this as a researcher there are other possible explanations for the resolution of the problems presented here which need to be explored. These outcomes may be explained by a number of different factors. For instance some individuals improve because they have entered therapy, regardless of the specific treatment: a variant of the placebo effect. Fascinating research has been conducted into the well-known phenomenon of the placebo effect with various medical conditions: headaches, pain reduction and even the visual effects of packaging in headache tablets [85, 86].

It could further be suggested that 'narrative smoothing' plays a part in resolving psychological issues, and many psychotherapists support this view [87–89]. It is believed that the process of reconstruction of the initial narration gives more control over the story and can change the patient's perception into something more positive; this underpins the central goals of therapy [87]. Rebecca's narrative was highly charged with specific negative experiences and relating this allowed a different subjective perception.

CBT might also be effective as a therapy in this instance as it similarly uses narration in therapy but treats the presenting symptoms rather than the cause. CBT has similarities with psychodynamic therapies but there are a number of different elements which are distinctive. CBT focuses on the way individuals think and act in specific circumstances and how emotional and behavioural problems may be overcome [90]. However, although CBT appears to be the preferred treatment for anxiety-based conditions, no theory/therapeutic action is without flaws, and a number of issues have been identified with this approach [34]:

In fact there is increasing concern regarding the relapse rate at follow-up sessions for those patients who have undergone symptom-based CBT [19, 90]. It treats the symptoms rather than the cause and as such this may only provide a short-term solution to the problem [16, 34].

#### **7.7 Longitudinal outcome**

Since Rebecca's therapy treatments in 2010 she has completed her Open University degree and now holds a Masters in Professional Voice Practice. She still enjoys performing and has regular engagements. Her dream of becoming a professional musician has been realised.

#### **8. Conclusion**

The aim of this chapter was to give more exposure to and greater understanding of two highly effective therapies for the rapid reduction of cognitive anxiety: CH and EMDR. The complexities of cognitive anxiety were discussed and the role that

### *Cognitive Hypnotherapy and EMDR: Two Effective Psychodynamic Therapies for the Rapid… DOI: http://dx.doi.org/10.5772/intechopen.101770*

dysfunctional memories, particularly implicit memories no longer consciously perceived, can exert on present-day experiences. The chapter reviewed CBT and the benefits of hypnosis as an adjunct to this therapy, now called CH, as well as comorbid conditions where CH has been an effective treatment. The background and theory of EMDR was documented discussing the role of disparate memories in EMDR practice. The procedures and protocols used in EMDR were highlighted before reviewing the clinical studies that have adopted EMDR as a treatment. The final section of the chapter documented the author's research using CH and EMDR, giving both quantitative and qualitative findings. Both the quantitative and qualitative research documented above (pp. 8–13) gave evidence for the effectiveness and rapid results of CH and EMDR in comparison with other complementary treatments where the main drawback appears to be the length of time required to effect positive change. This chapter further highlighted the important contribution that CH and EMDR have made to current research and in doing so has shown the need for further scientifically-based research into the complexities of anxiety and the role that implicit memories play in maintaining this.

In the UK the treatments of choice being advocated by the National Health Service (NHS) for PTSD and anxiety disorders still tend to be medication or CBT and as such there is no financial cost incurred by the patient. Where individuals experience no positive effects, or are dissatisfied with treatment, they are guided towards private therapists at their own financial cost. The medical profession are aware of CH and EMDR but as yet these treatments are not widely available to the general population, although EMDR is available for PTSD in the military domain. This state of affairs needs to be remedied given the effective and dynamic results which are being seen from both CH and EMDR.


*4.10. In Vivo Mouse Study for Screening of Antitumor Activity and Toxicity* 4.10.1. Mice Preparation and Ethical Approval

Thirty male Swiss albino mice (weight range equaled 25–30 g, 6–8 weeks of age) were purchased from VACSERA (Helwan, Egypt) and placed in groups of six in plastic cages. The experiment was done in the institutional animal house at the Faculty of Pharmacy, Suez Canal University, at a temperature range equal to 23 ± 5 ◦C, and the animals had free access to their normal diet and drinking water. The protocol of this study obtained approval from the institutional research ethics committee (#202004MA1).

## 4.10.2. Induction of Skin Lesions

A 2 × 2 cm2 dorsal skin area was shaved on all animals using a hair clipper 48 h prior to the experiment. To induce skin lesions in mice, one dose of DMBA, which acts as an initiator for skin tumors (100 µg in 200 µL acetone) [63], was injected subcutaneously into each mice [28]. After one week, there was an increase in the number of epidermal lesions (the lesions > 1 mm in diameter for each mouse [63]. The skin lesions were assessed first by skin morphology (lesion width and lesion length) and, also, by histological methods (thickness of the epidermis).

4.10.3. Regimen of Applying Metformin-Loaded Ethosomes

Each group contained 6 animals, and the selected optimal formula was applied topically on the dorsal region of the skin (10 mg/cm2 of the affected area) per week for a total of 4 weeks [27,48]. The experimental groups are shown in Table 4.

**Table 4.** Experimental groups for the in vivo mouse study.

The topical empty gel contained distilled water and carbopol 974 p only. On the other hand, the topical empty ethosome gel contained all the ethosome components (distilled water, ethanol, isopropyl alcohol, lecithin and cholesterol) and carbopol 974 p without metformin.

The measurements of the diameters of the skin cancer lesions were standardized for evaluating the efficacy of the selected optimal gel formula. Each mouse lesion more than 1 mm in diameter was measured weekly until the end of the study. At the end of the study protocol, the final lesion diameters were determined, and then, the mice were anaesthetized and slaughtered [64]. Blood samples were taken by cardiac puncture and settled for 30 min before centrifugation and separation of the serum samples.

#### 4.10.4. Histopathological Methodology and Examination

Extracted skin specimens were fixed in 10% neutral-buffered formalin, then embedded in paraffin wax and sectioned by a microtome (at 4 µm) and processed for hematoxylin and eosin (H&E) staining (the sections were deparaffinized, rehydrated in alcohol, stained in Harris hematoxylin, rinsed in 95% ethanol, counterstained with eosin solution, dehydrated through 95% alcohol and cleared in xylene, followed by mounting). Light microscopy was used to examine the skin sections by an experienced pathologist (the nuclei, nucleolus and nuclear membrane were stained blue, and the cytoplasm and connective tissue were stained pink) [30]. Histopathological investigation for skin tumor specimens was performed to assess the efficacy of the different drug/vehicle formulations [63]. Thicknesses of six regularly spaced skin parts were measured using ImageJ software (NIH, Bethesda, MD, USA). The average of the measured parts was calculated for every tissue specimen.

#### 4.10.5. Toxicological Screening

For testing any possibility of hepatic or renal toxicity due to the systemic absorption of the gel formula, a histopathological investigation was done for the liver and kidney specimens. The tissue samples were fixed in neutral-buffered formalin and processed for H&E staining and examination under light microscopy by an experienced blinded pathologist. In addition to the histopathological examination, the serum samples were directed for estimation of the liver enzymes (ALT and AST) and serum creatinine, urea and albumin.

#### *4.11. Statistical Analysis*

GraphPad prism was used to apply the statistical tests to the current data. Data were quantitative in nature and demonstrated in the form of the mean ± SD and analyzed using one-way ANOVA, as one factor (treatment regimen) was influencing the study groups. Bonferroni's test for multiple comparison analysis was at *p* < 0.05.

#### **5. Conclusions**

The topical application of ethosomal gel of metformin has a significant effect on treating chemically induced skin cancer in mice. This was shown by using the Box–Behnken design of "a three-level three-factor" to present a high percent of the EE%, minimum vesicle size, maximum ZP and high DR%.

Adding isopropyl alcohol with ethanol to form ethosomes increased the ability of ethanol to solubilize lecithin, which led to an increased stability and effectiveness of the ethosomes vesicles. Isopropyl alcohol also decreased the particle size of the vesicles, which increased the EE% and allowed the metformin to be released for an extended period. This is the goal of our research. Finally, a lower lecithin, high ethanol and isopropyl alcohol and moderate cholesterol obtained an enhancement of the permeation rate. Hence, the current findings may open up an avenue for future formulations for metformin as a therapeutic tool in fighting skin cancer.

**Author Contributions:** Conceptualization: O.M.S., T.M.H. and S.G.; Data Curation: I.A.M., T.M.H., S.G., S.A.Z., M.E.-S. and O.M.S.; Formal Analysis: I.A.M., S.A.Z. and O.M.S.; Software: I.A.M., T.M.H. and S.G.; Methodology: I.A.M. and O.M.S.; Resources: I.A.M. and O.M.S.; Validation: I.A.M. and O.M.S.; Visualization: T.M.H., S.G., S.A.Z., M.E.-S. and O.M.S.; Writing the Original Draft: I.A.M. and Writing, Review and Editing: T.M.H., S.G., S.A.Z., M.E.-S. and O.M.S. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research was funded partly by the AlMaarefa University researchers supporting program, AlMaarefa University, Riyadh, Saudi Arabia, grant number MA-006.

**Institutional Review Board Statement:** The animal study protocol was approved by the Ethics Committee of Faculty of Pharmacy, Suez Canal University (protocol code 202004MA1 on 12 April 2020).

**Informed Consent Statement:** Not applicable.

**Data Availability Statement:** Data is contained within the article.

**Acknowledgments:** The authors are thankful to Medical Union Pharmaceuticals (Abu Sultan, Egypt) for kindly providing the metformin hydrochloride powder, and also, we thank Future Pharmaceutical Company for providing the carbopol 974p powder.

**Conflicts of Interest:** The authors declare no conflict of interest.

#### **References**


**5. Conclusions**

 Medicinal and aromatic plants still have unknown and uncountable potential, though there is a long history for use since prehistoric times. To loose genetic resources leads a weighty loss in the future; therefore, the coexistence balance among plants, animal, and human should not be upset. The role of medicinal and aromatic plants is changing continuously in accord to a period and the role expands such as cure of disease to prevention of disease. The accumulated massive knowledge, information, and materials should be shared in the whole world and go down to generation to generation. The blessings of medicinal and aromatic plants are treasures that belong to all lives.

*Pharmacognosy - Medicinal Plants*


**2. Background**

Raw material demand has steadily increased on a global scale as a result of demographic and economic developments. If current trends in raw material use intensify, Industrial technologies will be unable to meet this growing demand. As a result, it is necessary to highlight that raw material (CDs present case) production must be supported as a strategic necessity, requiring the development of new technologies that can help meet this upcoming raw material demand. An additional challenge for the mining industry is that, as the world's mineral reserves are exhausted and demand for metallic raw materials rises, it will be designed to process ever greater amounts of low-quality mined material to manufacture concentrates in adequate quantities to meet current and potential demand. As a result of these requirements, minerals trapped in tailings ponds have started to receive interest as a potential source of raw materials, as many existing ore bodies near depletion. As a result, the mineral processing industry was interested in finding new solutions for the treatment of tailings to reprocess. It's worth remembering that the global amount of tailings was enormous, and if an available processing method could be developed, this could translate into a massive feed stream for the metals industry [11]. One of the most commonly used enrichment methods in mineral processing is froth flotation separation. The flotation separation is used in the processing of metals like copper, gold, and platinum to produce concentrates that can be refined economically. Efforts are currently being made to better understand the various phenomenon like frother phenomenon, adsorption etc. and how to regulate it, but no genuinely innovative frother method has been proposed, as most studies are focusing on CDs [12, 13]. As soon as synthetic polymers (CDs) were introduced into mineral processing, combinations with cellulosic materials became available and the importance of CDs came to light.

Selective depressants were essential components of any flotation reagent scheme that aims to separate various minerals selectively. Inorganic depressants have been used often. Many of these depressants, especially those used in differential sulphide flotation, are highly toxic and unsuitable for use in the environment. Sodium cyanide, sodium dichromate, sulfur dioxide, arsenic trioxide, phosphorous

*Application of Cellulose Derivatives in Mineral Processing DOI: http://dx.doi.org/10.5772/intechopen.97127*

pentasulfide, and other depressants were examples. Since some of these inorganic depressants were reducing agents, they can be oxidized in aerated flotation pulps. The use of a lot of reagents was normally the result. Polysaccharides, on the other hand, were non-toxic and biodegradable natural organic polymers. They're much less expensive and less prone to oxidation than inorganic depressants. These characteristics not only make them perfect flotation reagents, but they have also shown promise as selective depressants in a variety of differential mineral flotation systems. For nearly 70 years, cellulose derivatives (CDs) and polysaccharides/CDs have been used in the mineral industry as depressants for iron oxides, naturally hydrophobic minerals, and rock-forming gangue minerals. They've also been reported to be selective in sulphide mineral differential flotation [14].


Laser

In the stereolithography apparatus (SLA), a source of high-power laser solidifies a liquid resin. In a bath full of resin, the light source can produce the desired pattern [20,53]. Superior printability and cell encapsulation capacity have been reported in the biodegradable hydrogel construct in the study of Elomaa et al. [54]. Gaebel et al. [55] introduced cardiac patches for the treatment of myocardial infarction, where cardiac patch was seeded with human umbilical vein endothelial cells and human MSC, improved wound healing and functional preservation.

The main advantage of this method is the fidelity of the achieved geometries. Complex patterns can be manufactured with high resolution, like vascular networks with wide range scales (50–250 μm) [14,56]. The contactless procedure of laser-based bioprinting prevent cells from facing mechanical stresses; thus, high cell viability is expected [53].

The disadvantage of the method is the necessity of photosensitive materials, and hence the selection of bioniks is limited [14,23]. Furthermore, the prerequisite of incorporating a cell type into a hydrogel restricts other possible bioinks [23]. In addition, the cost of the laser diodes is usually higher than its counterparts (nozzles). Moreover, future research could reveal the side effects in cells after the laser exposure during the manufacturing procedure [53].

**Table 1.** Advantages and limitations of 3D bioprinting techniques for CVD treatment.

#### 2.2.2. Biomaterials and Inks


**3. Results**

### **3.1 Von Mises stress and the first principal stress**

**Figure 4** shows the results of von Mises stress in each computation. While the shape of the placenta was largely deformed because of the myometrium contraction and elongation, von Mises stress in the placenta was kept much lower than that in the chorionic and basal plates, marginal region, and myometrium. Also, von Mises stress at these layers fluctuated largely. **Figure 5** shows the results of the first principal stress. As the results of von Mises stress, the first principal stress in the placenta

#### **Figure 4.**

*von Mises stress in the computational model of the human placenta and surroundings, caused by the myometrium elongation and contraction (Mode 1, Mode 2, and Mode 3 (***Table 3***)). The stress distribution is shown by the entire image (right), and the values along each line, indicated by* **Figure 3** *(left).*

*Histological Properties of the Chorionic and Basal Plates and Maintenance of the Mechanical… DOI: http://dx.doi.org/10.5772/intechopen.112935*

#### **Figure 5.**

*The first principal stress in the computational model of the human placenta and surroundings, caused by the myometrium elongation and contraction (Mode 1, Mode 2, and Mode 3 (***Table 3***)). The distribution of the stress is shown by the entire image (right), and the values along each line, indicated by* **Figure 4** *(left).*

was kept lower. The chorionic and basal plates, and marginal region, whose elastic moduli were much higher than the placenta, would prevent changes in the mechanical environment of the placenta. The stable condition in von Mises stress and the first principal stress would promote the effective usage of the perivascular contractile cells in the villous tree and chorionic plate for the fetal and maternal blood circulations.

#### **3.2 Displacement and the first principal strain**

**Figure 5** shows the result of displacement in each computation. The deformation in the placenta at Mode 1 was much larger than that in other cases. The elongation and contraction the myometrium would cause various deformation patterns in the placenta so that the fetal and maternal blood could be stirred. As **Figure 6** shows, the first principal strain showed the same tendency (**Figure 7**).

#### **Figure 6.**

*Displacement in the computational model of the human placenta and surroundings, caused by the myometrium elongation and contraction (Mode 1, Mode 2, and Mode 3 (***Table 3***)). The distribution of the displacement is shown by the entire image (right), and the values along each line, indicated by* **Figure 3** *(left).*