**3. Antinutritional factors and microbial degradation**

Legumes contain several ANFs, such as raffinose, phytic acid, condensed tannins, saponins, alkaloids, lectins, pyrimidine glycosides, and protease inhibitors [17]. Overall, ANFs decrease the bioaccessibility and bioavailability of other nutrients, and, in some cases, are responsible for adverse reactions to the ingestion.

The content of raffinose-family oligosaccharides (RFOs, raffinose, verbascose, and stachyose) in legumes ranges from 1 to 6% with stachyose as the most abundant compound [18]. While in cereals, it is commonly lower than 1.5%, with raffinose as the sole or the most abundant compound [19, 20]. RFOs are nondigestible oligosaccharides that may result in adverse digestive symptoms when about 15 g/person per day are exceeded [21], a threshold that is readily reached in legume-based diets. Raffinose and RFO are indeed fermented by the intestinal microbiota with abundant gas production, causing discomfort and flatulence.

Phytic acid is the main storage compound for phosphorous and minerals in cereal and legume seeds. In legumes, its concentration can reach 20 g/kg [22, 23]. Phytic acid and divalent minerals (e.g., Ca2+, Zn2+ and iron) form stable complexes (phytates) that are insoluble and not hydrolyzed in the gastrointestinal tract, thus reducing the bioavailability of minerals for the monogastrics. Ca2+ and Zn2+ deficiencies are commonly observed in developing countries, and complexation of dietary minerals by phytates in plant-derived foods contributes to the mineral deficiency [17]. Iron uptake from plant-derived foods is impeded not only by complexation with phytate but also by complexation with condensed tannins [24, 25].

Proanthocyanidins, gallotannins. and ellagitannins, commonly referred to as tannins, are phenolic compounds that occur in a wide variety of plant foods. Their presence in cereals and legumes is dependent on the plant species and the cultivar [26]. Tannins impart bitter taste, reduce protein and starch digestibility by inhibition of pancreatic enzymes, and reduce iron uptake [26, 27]. The presence of tannins reduces the caloric content and the glycemic index of foods [28], but the abundance in diet reduces the supply of macro- and micro-nutrients.

Lectins and specific inhibitors of digestive enzymes (proteases and amylases) further reduce the digestibility of starch and proteins in legumes [26, 29].

Some ANFs are heat-labile (e.g., protease inhibitors and lectins) and easily removed by thermal treatments. Nevertheless, phytic acid, raffinose, tannins, and saponins are rather thermostable. Dehulling, soaking, air classification, extrusion, steaming, and pregelatinization are the main technological options for decreasing

*Fermentation as Strategy for Improving Nutritional, Functional, Technological, and Sensory… DOI: http://dx.doi.org/10.5772/intechopen.102523*

the negative impact of ANF on legume consumption [30–32]. Nevertheless, biological methods such as germination, enzyme treatments, and especially, fermentation seem to be more efficient [30, 31, 33, 34].

Proteolysis, enzyme inhibition due to acidification, acid activation of flour endogenous enzymes (e.g., phytases) and/or microbial enzyme activities (e.g., α-galactosidase, β-glucosidase, phytases, tannases) are responsible for the inactivation of most ANFs.

Raffinose family oligosaccharides are hydrolyzed through the activity of α-galactosidases, levansucrase, and sucrose-phosphorylase activities of lactic acid bacteria [35, 36] or corresponding enzymes of fungal cultures; their removal in legume fermentations has been amply reported [37].

In cereal matrices [22], the phytase activity is often sufficient to degrade phytates, especially in acidic conditions [18, 38]. Therefore, phytate degradation in LAB-fermented matrices spontaneously occurs without microbial enzymes involvement [18]. The optimal pH for the activity of the cereal phytases corresponds to 5.5; nevertheless, phytases are still active at pH levels lower than those commonly reached by sourdough (3.8–4.2) [18]. Sourdough fermentation and other types of traditional bioprocesses involving LAB (e.g., fermentations for production of cereal porridges or beverages) allow the increase of the mineral bioavailability [39]. Compared with that found in cereals, the phytase activity in legumes is poor [22, 40]. Nevertheless, pretreatments and processing conditions including fractionation, germination, soaking, thermal treatments, and fermentation drastically decrease phytate levels in legumes [41]. In many spontaneously fermented legume products, substrate-derived phytases are inactivated, and phytate degradation is achieved by fermentation with bacilli or fungal cultures, for example, *Rhizopus stolonifer* or *Aspergillus oryzae*, which hydrolyze phytate with extracellular enzymes [42, 43].

Metabolism of tannins or other polyphenols by LAB was deeply characterized only in a few fermented plant-derived matrices [44, 45]. *Lactiplantibacillus plantarum*, *Lactiplantibacillus paraplantarum*, *Lactiplantibacillus pentosus* have been identified, among the LAB, as the species that could decrease the tannins concentration through their tannases (tannin acyl hydrolase, EC <IP_ADDRESS>) [46–48]. However, characterization of fermented cassava allowed identifying uncommon tannase producers such as *Weissella cibaria* and *Leuconostoc mesenteroides* ssp. *mesenteroides* [49]. Most of tannase producers were found in fermented vegetables but also in human feces. In *L. plantarum*, tannase is very well characterized. Its activity was demonstrated and characterized by Rodríguez et al. [47], and genetic analysis showed that it constitutes a novel family of tannases [50]. LAB tannases are intracellular. Genes involved in tannins degradation are regulated in a coordinated way and are inducible by tannin and other phenolic compounds [51].

The lactic fermentation of grass pea (*Lathyrus sativus*) with *L. plantarum* lowered the levels of phytic acid and trypsin inhibitory activity [52]. Selected strains of *L. plantarum* and *Levilactobacillus brevis* decreased the content of raffinose up to circa 64% during sourdough fermentation of different legume flours. Sourdoughs made with different legume flours (bean, lentil, pea, grass-pea, chickpea) contained an increased phytase activity compared with the unfermented controls [34]. The combination of legume sprouting and sourdough fermentation decreased the content of phytic acid, condensed tannins and raffinose, and trypsin inhibitory activity [53, 54].

Besides the abovementioned ANFs, faba bean is rich in two glucosidic aminopyrimidine derivatives, vicine and convicine, which, upon hydrolysis of the β-glucosidic bond, generate the aglycones divicine (2,6-diamino-4,5-dihydroxypyrimidine) and isouramil (6-amino-2,4,5-trihydroxypyrimidine), respectively [55]. Divicine and isouramil trigger favism disease in susceptible individuals. Technological processes (air classification, roasting, and boiling) and selection of cultivars with low content of such compounds seemed to be only in part effective [55, 56]. On the contrary, β-glucosidase from LAB effectively degraded the pyrimidine glycosides from faba bean suspension and flour [30]. When used as starter to ferment fava bean flour, *L. plantarum* expressed β-glucosidase activity and decreased the content of vicine and convicine by more than 90%. The degradation was complete after 48 h of fermentation, and aglycone derivatives were not detectable [57]. Similar results were obtained when flours from different faba bean accessions collected from the Mediterranean area were subjected to the LAB fermentation [58]. *Ex-vivo* hemolysis assays on human blood confirmed the lack of toxicity of the fermented fava bean [57].


**5. The nano alpha-X instrument for MUSES-C mission**

In the year of 2000, NASA, with collaboration with Japanese JAXA, were getting ready for the MUSES-C mission to bring back to Earth a sample from an asteroid. On that mission, it was also proposed to study in-situ the surface characteristics of that asteroid with a nanorover populated with several analytical instruments, including an Alpha-X Spectrometer (AXS) [22] to obtain the chemical composition of the asteroid surface material. For that mission, a further miniaturization of the AXS was required to fit it inside the Jet Propulsion Laboratory's nanorover with the overall dimensions of 14x15x6 cm. A massive hybridization and elimination of the proton mode resulted in a miniaturized Alpha-X Spectrometer shown in Fig. 12 and with specifications listed in Table 3. The AXS, although drastically miniaturized, is a complete spectrometer capable of providing highly accurate analytical results, similar to other APXS instruments.

Fig. 12. A photograph of AXS laboratory instrument for MUSES-C mission.

In the year of 2000, NASA, with collaboration with Japanese JAXA, were getting ready for the MUSES-C mission to bring back to Earth a sample from an asteroid. On that mission, it was also proposed to study in-situ the surface characteristics of that asteroid with a nanorover populated with several analytical instruments, including an Alpha-X Spectrometer (AXS) [22] to obtain the chemical composition of the asteroid surface material. For that mission, a further miniaturization of the AXS was required to fit it inside the Jet Propulsion Laboratory's nanorover with the overall dimensions of 14x15x6 cm. A massive hybridization and elimination of the proton mode resulted in a miniaturized Alpha-X Spectrometer shown in Fig. 12 and with specifications listed in Table 3. The AXS, although drastically miniaturized, is a complete spectrometer capable of providing highly accurate

**5. The nano alpha-X instrument for MUSES-C mission**

analytical results, similar to other APXS instruments.

Fig. 12. A photograph of AXS laboratory instrument for MUSES-C mission.

Table 3. Characteristics of the AXS instrument for MUSES-C mission.

History of Applications of Radioactive Sources in Analytical Instruments for Planetary Exploration 171

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[3] Patterson, J.H, A. L. Turkevich and E. J. Franzgrote, Chemical analysis of surfaces using

[4] Thanasis E. Economou, Anthony L. Turkevich, Keneth P. Sowinski, James H. Patterson

[5] Thanasis E. Economou,Anthony L. Turkevich and James H. Patterson, An Alpha Particle

[6] Patterson, J.H, Ernest J. Franzgrote,A. L. Turkevich, W.A. Anderson, T.E. Economou,

[7] James H. Patterson, Harry E. Griffin, E. Philip Horwitz, and Carol A. Bloomquist,

[8] Thanasis E. Economou and Anthony L. Turkevich. "An Alpha Particle Instrument with

[9] Hovestadt, D., et al., PHOBOS, Proceedings of the International Workshop, Moscow,

[10] Economou, T.E, J.S. Iwanczyk and R. Rieder, A HgI2 X-ray Instrument for the Soviet

[11] Linkin, V., et al., "A Sophisticated Lander for Scientific Exploration of Mars :

[12] Rieder, R., H. Wänke, T. Economou and A. Turkevich; "Determination of the Chemical

[13] Rieder, R., T. Economou, J. Brückner, G. Dreibus, H. Wänke and A. Turkevich, First

[14] Rieder, R., T. Economou, H. Wänke, A. Turkevich, J. Crisp, J. Brückner, G.

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Crotch, J. H. Patterson and K. P. Sowinski. "The Alpha-Scattering Chemical Analysis Experiment on the Surveyor Lunar Missions". Jet Propulsion Laboratory

and Ernest J. Franzgrote, The Alpha-Scattering Technique of Chemical Analysis, J.

Experiment for Chemical Analysis of the Martian Surface and Atmosphere, J.

H.E. Griffin, S.L. Groach and K.P. Sowinski, Alpha Scattering Experiment on Surveyor 7: Comparison with Surveyor 5 and 6, J. Geophys. Res., 74, 6120,

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Alpha, Proton and X-ray Modes for Planetary Chemical Analyses". Nucl. Instr. &

Scientific Objective and Implementation", Planetary Space Science Journal, Vol. 46,

Composition of Martian Soil and Rocks: The Alpha-Proton-X-Ray Spectrometer", J.

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Dreibus,H.Y. McSween, Jr., The Chemical Composition of Martian Soil and Rocks Returned by the Mobile Alpha Proton X-ray Spectrometer: Preliminary Results

charged particles", Science, 134, 672, 1961.

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**7. References**

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1986, p 302

The MUSES-C mission suffered several long delays and NASA eventually cancelled its participation on this mission. The Japanese, however, went ahead with the mission that was renamed Hayabusa after the launch in 2003 and succeeded in bringing back a small amount of asteroid material from asteroid Itokawa in June, 2010. Despite the cancellation, the AXS was designed, built and fully flight qualified. It is now available for some potential future mission.


**3. The pathophysiological mechanisms of viral myocarditis**

The current understanding of the immunopathologcial phases of myocarditis derives largely from enterovirus-induced myocarditis in murine models (Andréoletti et al, 1997; Cooper et al., 2007; Liu & Mason, 2001; Yajima & Knowlton, 2009). During the enterovirus (virus Coxsackie B) natural course of infection, the data obtained from experimental murine

Common human pathogenic DNA or RNA viruses can be responsible for acute or chronic endomyocardial tissue infection (Andréoletti et al., 1995; 2009; Kühl et al., 2005a). The detection of components (DNA or RNA genomes or viral proteins) of these viral agents by molecular techniques such as polymerase chain reaction (PCR) and/or immunohistochemical techniques demonstrating the viral protein expression in the cytoplasm of myocytes, was associated with an inflammation of the myocardium (acute myocarditis), arrhythmias, loss of contractility (Andréoletti et al., 2009; Cooper, 2009; Dennert et al., 2008). Moreover in serial of human cases, the presence of viral persistence was evidenced in the myocardium and associated with a left ventricular systolic dysfunction in relation to a reduction in the contractile function of the myocytes (Andréoletti et al., 2000; Badorff et al., 2001). Additional proofs of concept were that: (I) a longitudinal clinical investigation showed that the type of virus, the clinical presentation and the type of histological damage appeared to be related to the clinical course of the cardiac disease; (II) longitudinal clinical studies indicated that the immune clearance of the virus during or after the acute phase was correlated with an improvement of the left ventricular ejection fraction [LVEF] (Kühl et al. 2005b); (III) acute and chronic myocarditis as well as dilated cardiomyopathy (DCM) were reproduced in immunocompetent animal models following experimental viral infections (Andréoletti et al., 1997; Huber, 1993; Matsumori & Kawai, 1982). Altogether these clinical and experimental data supported the pathophysiological role of several human viruses in the genesis and the evolution of myocarditis and the DCM. Although up to 90% of people will be infected by at least one or more of these viruses in their life without getting their heart injured with associated clinical signs, only a few number will develop clinical symptoms. Therefore, it is highly suspected that a certain genetic background either related to immune alterations or to an improved susceptibility to viral cardiac infection (viral receptor or co-receptors polymorphisms) may be a prerequisite to develop clinical symptoms of myocarditis and/or progression to DCM following virus heart tissue infection. In addition, it has been shown that dystrophin mutations may make easier the development and progression of myocarditis and cardiac failure during coxsackievirus B3 infection, whereas dystrophin and/or sarcoglycan disruption by viral proteases account for myocardial injury (Andréoletti et al., 2007; Badorff et al., 2001; Lee et al., 2000). Human genetic studies of patients with myocarditis are rare and only 2 reports show an association between myocarditis and genetic factors such as HLA-DQ locus and CD45 polymorphism (Yajima & Knowlton, 2009). Future clinical investigations should therefore focus on the understanding of the underlying genetic susceptibility and related immune responses that explain why some patients are susceptible to develop clinical symptoms of acute myocarditis following viral infection, whereas other subjects clinically recover or progress to an 'idiopathic' DCM after the initial phase of the viral cardiac

**2. Human viral cardiac infection: proofs of concept and clinical relevance**

infection.

**3. The pathophysiological mechanisms of viral myocarditis**

The current understanding of the immunopathologcial phases of myocarditis derives largely from enterovirus-induced myocarditis in murine models (Andréoletti et al, 1997; Cooper et al., 2007; Liu & Mason, 2001; Yajima & Knowlton, 2009). During the enterovirus (virus Coxsackie B) natural course of infection, the data obtained from experimental murine models allowed the identification of three distinct pathophysiological phases defined as acute, sub-acute and chronic myocarditis (Cooper, 2003; Feldman & McNamara, 2000; Yajima & Knowlton, 2009) (Figure 1).

Fig. 1. Immunopathological phases of viral myocarditis.