**2. Oxidative stress and multiple sclerosis**

OS is a cellular state where the homeostasis of redox reactions is altered when the production of reactive oxygen (ROS) and reactive nitrogen species (RNS) exceed their elimination. These reactive species are generated, among other causes, by oxidative metabolism. Neurons of the CNS are very active in oxidative metabolism, as they are constantly exposed to low‐to‐ moderate levels of ROS, and these species are removed by antioxidants (melatonin, vitamin D, vitamin E, glutathione) and antioxidant enzymes (superoxide dismutase, catalase, gluta‐ thione peroxidase, etc.). In chronic inflammatory diseases, such as MS, antioxidant defenses are overcome, which leads to oxidative stress [3].

Collectively, the ROS are reactive species derived from oxygen that include the superoxide anion (O- ), hydrogen peroxide (H2O2), and the hydroxyl radical (•OH). The RNS are reactive species derived from nitrogen and include nitric oxide (NO**∙**) and peroxynitrite (ONOO- ). The ROS and RNS are extremely unstable and reactive because they have an unpaired electron in their outer orbital. They take electrons from proteins, lipids, carbohydrates, and nucleic acids, causing damage to biological membranes, genetic material, and other macromolecules. The CNS is particularly vulnerable to oxidative damage since it has a very active mitochondrial metabolism, which leads to high levels of intracellular superoxide anions. Moreover, oligo‐ dendrocytes have low levels of antioxidant enzymes and a high concentration of iron. Unsaturated fatty acids are the most vulnerable to free radicals, and because myelin has a high lipid‐to‐protein ratio, it is a preferred target of ROS [4]. The ROS are generated by a number of cellular oxidative and metabolic processes including activity of the enzymes of the mito‐ chondrial respiratory chain, xanthine oxidase, NADPH oxidase, monoamine oxidases, and metabolism of arachidonic acid (AA) mediated by the activity of lipoxygenases (LOX), and ROS are produced primarily by leakage of electrons in the mitochondrial respiratory chain [3].

**Figure 2.** Oxidative stress levels are directly related to the progression of MS. Pathway Builder Online Tool was used to draw the figure [27].

Numerous studies in MS patients have shown an increase in the production of OS markers (such as cholesteryl ester hydroperoxides) and lower levels of uric acid (a ONOO scavenger). These changes are accompanied by significant deficiencies in antioxidant enzymes compared to healthy subjects. The increase in ROS coupled with decreased antioxidant capacity is not enough to entirely explain the pathogenesis of MS [4, 5]. Other reports suggest that the loss of myelin nerve sheath is possible because the immune system participates in combination with defects in the mitochondria, and these defects cause the generation of ROS and RNS. Macro‐ phages and monocytes release mediators of OS that degrade the unsaturated fatty acids. The ROS have also been implicated as a mediator of demyelination of axonal damage in MS and experimental autoimmune encephalomyelitis (EAE) [6]. It is important to mention that in assessing platelets in MS patients, increased activity of free radicals with decreased levels of important antioxidants such as glutathione and alpha‐tocopherol has been reported [7] (**Figure 2**).

moderate levels of ROS, and these species are removed by antioxidants (melatonin, vitamin D, vitamin E, glutathione) and antioxidant enzymes (superoxide dismutase, catalase, gluta‐ thione peroxidase, etc.). In chronic inflammatory diseases, such as MS, antioxidant defenses

Collectively, the ROS are reactive species derived from oxygen that include the superoxide

ROS and RNS are extremely unstable and reactive because they have an unpaired electron in their outer orbital. They take electrons from proteins, lipids, carbohydrates, and nucleic acids, causing damage to biological membranes, genetic material, and other macromolecules. The CNS is particularly vulnerable to oxidative damage since it has a very active mitochondrial metabolism, which leads to high levels of intracellular superoxide anions. Moreover, oligo‐ dendrocytes have low levels of antioxidant enzymes and a high concentration of iron. Unsaturated fatty acids are the most vulnerable to free radicals, and because myelin has a high lipid‐to‐protein ratio, it is a preferred target of ROS [4]. The ROS are generated by a number of cellular oxidative and metabolic processes including activity of the enzymes of the mito‐ chondrial respiratory chain, xanthine oxidase, NADPH oxidase, monoamine oxidases, and metabolism of arachidonic acid (AA) mediated by the activity of lipoxygenases (LOX), and ROS are produced primarily by leakage of electrons in the mitochondrial respiratory chain [3].

**Figure 2.** Oxidative stress levels are directly related to the progression of MS. Pathway Builder Online Tool was used

species derived from nitrogen and include nitric oxide (NO**∙**) and peroxynitrite (ONOO-

), hydrogen peroxide (H2O2), and the hydroxyl radical (•OH). The RNS are reactive

). The

are overcome, which leads to oxidative stress [3].

anion (O-

152 Trending Topics in Multiple Sclerosis

to draw the figure [27].

**Figure 3.** Damage to axons. Pathway Builder Online Tool was used to draw the figure [27].

The molecular mechanisms proposed to explain how ROS could specifically mediate brain damage are the following: (1) The lower levels of antioxidants can promote increased activity of lipoxygenase in CNS stimulating leukotriene production, thereby increasing the immu‐ noinflammatory processes in the cerebral cortex; (2) the damage to myelin can be caused by activation of T cells that may be activated for the presence of free radicals produced by the synthesis route of AA. Then appear the markers of OS associated with reduced activity of superoxide dismutase and the increase in glutamine, followed by increases of ∙OH and the production of peroxides which ultimately has a negative impact on myelin. After that, the evident changes in mitochondrial activity and finally changes in membrane fluidity (particu‐ larly, mitochondrial membranes) appear [8].

**Figure 4.** Effect of nitric oxide in inflammation. Pathway Builder Online Tool was used to draw the figure [27].

Paraclinical studies have shown an increased metabolism of the RNS in serum, lymphocytes, and cerebrospinal fluid of MS patients, which correlate to pathology studies. ONOO is also closely associated with acute inflammatory lesions [9]. Damage to axons is mediated by the following: (1) failure in mitochondrial energy metabolism due to inhibition of the respiratory chain by nitric oxide, which in turn causes a decrease in Na+ /K+ ATPase activity and alters Na + ‐dependent glutamate transporters, (2) over‐expression of glutamate receptors, (3) oligoden‐ droglial excitotoxicity, (4) massive influx of extracellular Ca++, (5) activation of proteases, and (6) impaired axonal transport. These mechanisms produce glutamate excitotoxicity and increased generation of nitric oxide leading to nitrosative stress.  When nitric oxide is combined with the superoxide anion, it generates a potent free radical, the pro‐oxidant peroxynitrite. Glutamate in turn causes neurodegeneration through the AMPA and NMDA receptors in oligodendrocytes and astrocytes (**Figure 3**). It is possible to explain the role of mediators using an experimental model of autoimmune encephalitis: Protection against the experimental disease occurs after administration of a glutamate antagonist [10].

Under physiologic conditions, nitric oxide is produced from L‐arginine by constitutive nitric oxide synthase (cNOS) and participates in a variety of important biological functions such as immunoregulation of inflammatory reactions, the downregulation of tumor necrosis factor (TNF)‐α production, MHC II expression in macrophages, induction of apoptosis in CD4 cells, physiological regulation of the mitochondrial respiratory chain, inhibition of antigen presen‐ tation, and leukocyte adhesion and migration. However, during inflammatory reactions, exposure of macrophages to interferon (IFN)‐γ and TNF‐α results in the activation of the inducible isoenzyme of NOS (iNOS), which increases up to 10 times the levels of nitric oxide. Nitric oxide facilitates the formation of peroxynitrite radicals. Only cells capable of generating a high flow of NO• have the potential for causing nitrosative stress. The role of nitric oxide in MS is therefore complex, and in fact, peroxynitrite is definitely more toxic than nitric oxide [9] (**Figure 4**).


**12.4 Pitting corrosion**

Bifilms can act as invasive pathways for corrodents into the interior of metals. The outside surface of a metal may be tolerably resistant to corrosion, but at the location at which a bifilm emerges, breaking the surface, the ingress of rain or salt water is likely to form an etch pit. The localized corrosion around the bifilm may be enhanced by precipitates of second phases and intermetallics which favor the wetted exterior surface of the bifilm (its wetted exterior surface contrasts with its dry, unbonded inner interfaces). These different compounds with different electrochemical potentials attached to the exterior surface of the bifilm can provide vigorous corrosion couples.

**Figure 8** shows a typical etch pit. Although the conventional explanation of the image would be that the etch pit has initiated the formation of cracks, the reverse is true. The cracks are bifilms, as can be identified from their morphology and precipitates. They have initiated the etch pit.

In the past decade there have been at least three, perhaps four or more, helicopter crashes, some extremely tragic, in which items of the drive train appear to have failed by fatigue initiated from an etch pit. Experts from around the world have been puzzled because an etch pit was far too small to have initiated the fatigue crack. In the case of one main rotor shaft, which appeared to have failed in this way, the shaft was designed with a safety factor of five. It is not conceivable that such a robust shaft could be threatened by an etch pit.

It is easily appreciated, that the etch pit is merely the witness to the presence of a bifilm crack. Furthermore, the bifilm could have been extensive, such as possibly extending over a major portion of the shaft. The shaft was formed, of course, from VAR steel, so that the probability of its being pre-cracked is virtually certain. The crack

**Figure 8.** *Etch pit in a steel turbine blade. Courtesy Metallurgical Associates Inc.*

would have evaded detection because, being formed by oxidation in vacuum, its oxide films would have been extremely thin. Also, as a universal feature of castings, and heat-treated products, especially if quenched, the interior is in tension, but the exterior surfaces are in compression. The crack on the outside of the shaft would therefore have been tightly closed.

Attempts to find bifilms by nondestructive testing (NDT) has proven to be tragically unreliable. As always in such difficulties, the clear way forward is to use only those processes which do not generate bifilms and which are therefore intrinsically reliable.

#### **12.5 Stress corrosion cracking (SCC)**

This dangerous failure mode involves almost no loss of metal by corrosion but can generate deep cracks by a time-dependent advance, often under only low stress. A metal can be sensitized to SCC by heat treatments.

There seems to be good evidence that SCC is a bifilm phenomenon, whereby the corrodent is simply moving through the 'air gaps' of the bifilms, linking bifilms by corrosion, driven by the stress concentration at the bifilm linkages [2].

The action of certain heat treatments to enhance SCC susceptibility is here proposed to arise from the precipitation of second phases on the bifilm. The favored formation of precipitates on bifilms seems to be the result of the reduction in the strain energy of formation, because the volume change and shape change of the new arrival can be more easily accommodated by the 'air gap' of the bifilm. The movement of part of the new phase into the air gap is likely to assist the forcing open of the air gap, so that percolation of the corrodent is facilitated [2].

#### **12.6 Hydrogen embrittlement (HE)**

There are numerous theories which have attempted to explain HE, but the phenomenon cannot yet be claimed to be clearly understood. In practice, the ingress of hydrogen into a stressed steel can result in gradual loss of ductility, and final fracture. The process has been identified as the slow progress of a crack until the final fracture when there is insufficient area to support the load. Hydrogen enters the steel as a proton released from certain corrosion mechanisms. For research purposes, hydrogen is introduced by electrochemical processes. Significantly, researchers complain about the interference of blistering which upsets their experiments during the charging processes, and report they are at a loss to know how the blisters can nucleate [2].

Once again, the bifilm seems more than adequate to explain all these observed characteristics of HE. The blisters are the observation of bifilms, inflated by hydrogen, near to the surface of the metal. Clearly, bifilms in the interior of the metal will also be experiencing the pressurization of hydrogen gas. Bifilms will almost certainly aid the progress of the gas into the interior of the metal, greatly accelerating the apparent rate of diffusion. However, it is probably mainly those isolated bifilms whose internal pressurization is leading to internal stress build up which is countering the ability of the metal to withstand tension.

There has been much interest in the attempts to desensitize a metal to HE by providing sinks for hydrogen. The sinks have been generally thought to be dislocations, and stress fields around carbide precipitates. This author has proposed that bifilms are probably significantly more capacious sinks, and the action of carbides is to precipitate onto bifilms and to prize them open, enabling them to accommodate even more hydrogen. He suggests that in an increasing supply of hydrogen, the bifilm would act as a temporary reduction in the deleterious effect of hydrogen, but

**17**

**Author details**

John Campbell

Birmingham, UK

Department of Metallurgy and Materials, Faculty of Engineering, The University of

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

\*Address all correspondence to<EMAIL_ADDRESS>
provided the original work is properly cited.

greatly valued by the engineering world.

*Perspective Chapter: A Personal Overview of Casting Processes*

this benefit would only exist at low hydrogen levels. When the hydrogen pressure in the bifilms equalled and the exceeded the yield stress, the damaging effects of HE

1.The entrainment of the oxide film on the liquid metal during casting processes leads to widespread damage to metals. Pre-cracking by a poor casting technique is central to the loss of properties, and to numerous failure modes, including those during solidification, during cooling to room temperature,

3.Casting processes using gravity pouring can be designed to yield significant benefits in bifilm reduction and are recommended if counter-gravity cannot

4.Ultimately, counter-gravity casting is strongly recommended to be the new

5.The current use of VAR steels in all critical applications (especially such applications as helicopter drive chains) appears to be dangerously unreliable.

6.The reliable secondary remelting process could be ESR if combined with a reliable electrode. The implementation of this process combination would be

7.Both primary and secondary casting processes can now be made to deliver

during metal processing, and during service conditions.

2.Casting processes involving top pouring are especially damaging.

casting norm, capable of delivering defect-free cast products.

economic metals which cannot fail; metals we can trust.

*DOI: http://dx.doi.org/10.5772/intechopen.93739*

would resume unchanged [2].

be provided [3].

**13. Conclusions**

this benefit would only exist at low hydrogen levels. When the hydrogen pressure in the bifilms equalled and the exceeded the yield stress, the damaging effects of HE would resume unchanged [2].