**Chromofungin**

190 Antimicrobial Agents

These new AMPs are integrated in the concept that highlights the key role of the adrenal medulla in the immunity (Sternberg, 2006) as previously reported for adrenaline and neuropeptide Y that regulate immunity systemically once released from the adrenal medulla. Furthermore, the adrenal medulla contains and releases large amounts of IL-6 and TNF-alpha in response to pro-inflammatory stimuli such as LPS, IL-1 alpha and IL-1 beta (Metz-Boutigue et al., 1998). The discovery of the presence of TLRs on the adrenal cortex cells raises the interesting possibility that the adrenal gland might have a direct role in the response to pathogens, activation of innate immune response and clearing of infectious

Several new antimicrobial peptides isolated from the granules of chromaffin cells of the bovine adrenal medulla correspond to CGA-derived peptides (Figure 3). The corresponding sequences are highly conserved in human. Interestingly, the main cleavage site in position 78-79 of bCGA and the subsequent remove of the two basic residues K77 and K78 by the carboxipeptidase H (Metz-Boutigue et al., 1993) produces two antimicrobial fragments: vasostatin-I (VS-I; bCGA1-76) (Lugardon et al., 2000) and prochromacin (Prochrom; bCGA79-431) (Strub et al.,1996b). For these N- and C-terminal domains with antimicrobial activities several shorter active fragments were identified: for VS-I, bCGA1-40 (N CgA; NCA) (Shooshtarizadeh et al., 2010), bCGA47-66 (chromofungin; CHR) and for ProChrom, bCGA173-194 (Chromacin; Chrom) (Strub et al., 1996b) and bCGA344-364 (Catestatin; CAT) (Shooshtarizadeh et al., 2010). The unique disulfide bridge of bCGA is present in VS-I and NCA sequences. Two post-translational modifications are important for the expression of the antibacterial activity of Chrom: the phosphorylation of Y173 and the O-glycosylation of S186 [130] (Strub et al., 1996a). Furthermore, it is important to point out that a dimerization motif GXXXG similar to that reported for Glycophorin A (Brosig & Langosch, 1998) is

Vasostatin-I (VS-I) displays antimicrobial activity against (i) Gram-positive bacteria (*Micrococcus luteus* and *Bacillus megaterium*) with a minimal inhibitory concentration (MIC) in the range 0.1-1 µM; (ii) against filamentous fungi (*Neurospora crassa, Aspergillus fumigatus, Alternaria brassicola, Nectria haematococca, Fusarium culmorum, Fusarium oxysporum*) with a MIC of 0.5-3 µM and (iii) against yeast cells (*Saccharomyces cerevisiae, Candida albicans*) with a MIC of 2 µM (Lugardon et al., 2000). However VS-I is unable to inhibit the growth of *Escherichia coli* SBS363 and *Escherichia coli* D22. VS-I (Figure 3) possesses structural features specific for antimicrobial peptides, such as a global positive charge (+3), an equilibrated number of polar and hydrophobic residues (20:23) and the presence of a helical region CGA40-65 characterized to be a calmodulin-binding sequence (Lugardon et al., 2001; Yoo, 1992). The loss of the antibacterial activity of CGA7-57 suggests that the N- and C-terminal sequences are essential, nevertheless CGA7-57 is less efficient than VS-I against fungi. Besides, the disulfide bridge is essential for the antibacterial, but not the antifungal property. Altogether, these data suggest that antibacterial and antifungal activities of VS-I have different structural requirements (Lugardon et al., 2001). Interestingly, two helix-helix dimerization motifs important for the interaction with membranes such as LXXXXXXL,

agents (Sternberg, 2006).

**2.2 Antimicrobial peptides derived from chromogranin A**

present in the Chrom sequence (G184-G188).

**Vasostatin-I**

When VS-I was treated with the endoprotease Glu-C from *Staphylococcus aureus*, one of the generated peptide, chromofungin (CHR), is the shortest active VS-I-derived peptide with antimicrobial activities (Figure 3). It is well conserved during evolution and displays antifungal activity at 2-15 µM against filamentous fungi (*Neurospora crassa, Aspergillus fumigatus, Alternaria brassicola, Nectria haematococca, Fusarium culmorum, Fusarium oxysporum*) and yeast cells (*Candida albicans, Candida tropicalis, Candida neoformans*) (Lugardon et al., 2001). Since this peptide was generated after digestion of the material present in chromaffin secretory vesicles by the endoprotease Glu-C from *S. aureus*, it may be hypothesized that it is produced during infections by this class of pathogens.

The 3-D structure of CHR has been determined in water-trifluoroethanol (50:50) by using 1H-NMR spectroscopy. This analysis revealed the amphipathic helical structure of the sequence 53-56, whereas the segment 48-52 confers hydrophobic character (Lugardon et al., 2001).The importance of the amphipathic sequence for antifungal activity was demonstrated by the loss of such activity against *N. crassa* when two proline residues were substituted for L61 and L64, disrupting the helical structure, the amphipathic character and the dimerization motif helix-helix L57-L64 (Lugardon et al., 2001).

### **Catestatin**

Two CGA-derived fragments bCGA333-364 and bCGA343-362 were characterized after the extensive processing of bCGA by prohormone convertases (PC 1/3 or 2) in chromaffin granules (Taylor et al., 2000). More recently, it was shown that cathepsin L colocalizes with CGA in chromaffin granules*. In vitro* it is able to generate after digestion of recombinant hCGA, a catestatin (CAT)–derived fragment hCGA360-373 (Biswas et al., 2009). In addition to the inhibitory effect of CAT on catecholamine release from chromaffin cells (Mahata et al., 1997), we have shown for this peptide and its shorter active sequence bCGA344-358 (cateslytin, CTL), (Figure 3) a potent antimicrobial activity with a MIC in the lowmicromolar range against Gram-positive bacteria (*Micrococcus luteus, Bacillus megaterium* at concentration of 0.8 µM), Gram-negative bacteria (*Escherichia coli D22* at concentration of 8 µM), filamentous fungi (*Neurospora crassa, Aspergillus fumigatus, Nectria haematococca* at concentration of 0.2-10 µM) and yeasts (*Candida albicans, Candida tropicalis, Candida glabrata, Candida neoformans* at concentration of 1.2-8 µM). The sequence of CAT (Figure 3) has been highly conserved during evolution (Briolat et al., 2005). The two human variants P370L and

The Natural Antimicrobial Chromogranins/Secretogranins-Derived

bacteria.

Peptides – Production, Lytic Activity and Processing by Bacterial Proteases 193

of bacterial proteases on the isolated AMPs derived from CGs, we have tested the effects of *Staphylococcus aureus* V8 protease Glu-C and several supernatants from *S aureus*, *Salmonella enteretica*, *Klebsiella oxytoca*, *Shigella sonnei* and *Vibrio cholera*. By using biochemical methods we have analyzed the degradation of the peptide in presence of

*Interaction of antimicrobial CGs-derived peptides with proteases from diarrheogenic bacteria* Bacteria were isolated from patients of the Strasbourg Civil Hospital by the Bacteriology Institute, University of Strasbourg, (EA-4438). The four strains have a clinical interest

Thus, *Klebsiella* was involved in the occurrence of post-antibiotic diarrheas (Gorkiewicz 2009). Many studies show that *Klebsiella oxytoca* is also involved in nosocomial infections for newborns or adults (Biran et al., 2010) *Klebsiella* infections may also be commensal (Tsakris et al., 2011). *Klebsiella oxytoca* has also been associated with hemorrhagic colitis (Hoffmann et al., 2010) and intercurrent colitis in Crohn's disease (Plessier et al. 2002). *Salmonella* destroys infected cells and the infection continues through blood (sepsis) or through lymphatic vessel (typhoid fever). Salmonella cause also gastrointestinal infections. *Shigella sonnei* and *Vibrio cholera* non O1 cause inflammation of the intestinal

*Klebsiella oxytoca, Salmonella enterica, Shigella sonnei, and Vibrio cholera* develop phenomena of antibiotic resistance. Thus, *Salmonella* was reported to be resistant for the action of

Concerning CgA, we have tested bovine, rat and human CAT corresponding to the sequences bCgA344-364, rCgA6344-364 and hCgA352-372, bovine CTL located at bCgA344- 358, two short fragments hCgA360-372 and the conserved tetrapeptide LSFR (bCgA348-351). In addition, we have tested a scrambled peptide relative to the sequence of bovine CAT and

We have found antimicrobial activities only for the bovine CAT and CTL, showing that CTL is the shorter active fragment and that it corresponds to the active domain of CAT. Procatestatin was inactive in similar experimental conditions. Bovine CAT and CTL were active against *Klebsiella oxytoca*, *Salmonella enterica* and *Vibrio cholera* at 100 µM and 50 µM respectively and against *Shigella Sonnei* at 50 µM and 25 µM. In addition, CHR and the Cterminal fragment (CgA387-431) were inactive for concentration up to 100 µM. In contrast,

Three CgB-derived peptides (CgB58-62, CgB279-291, and CgB547-560) and secretoneurin corresponding to SgII189-254 were examined against the four strains in order to analyse their degradation by bacterial proteases. By using HPLC we have compared the profiles of

These experiments show that except CTL all the peptides are completely degraded by the bacteria. To illustrate these data, we present on Figure 4, the profiles relative to CAT and CTL in presence of buffer with *Salmonella enterica*. The complete peptide and the processed

because apart from inducing diarrhea, they may cause other infections.

Ciprofloxacine (Medalla et al., 2011) and Ceftriaxone (Su et al., 2011).

mucosa by producing the Shiga toxin.

the procatestatin fragment bCgA332-364.

CTL is active at 30 µM against the four pathogens.

the peptide alone and the peptide with the inoculated medium.

form are analysed by sequencing and mass spectrometry (MALDI-TOF).

G364S display antibacterial activity against *M. luteus* with a MIC of 2 and 1 µM, respectively, and against *E. coli* with a MIC of 20 and 10 µM, respectively (Briolat et al., 2005). However, the most active peptide corresponds to the bovine sequence. Bovine CTL, a cationic sequence with a global net charge of +5 (R344, R347, R351, R353, R358) and five hydrophobic residues (M346, L348, F360, Y355, F357) (Figure 3), is able to completely kill bacteria at concentration lower than 10 µM even in the presence of NaCl (0-150 mM) (Briolat et al., 2005). The C-terminal sequence bCGA352-358 is inactive, whereas the N-terminal sequences bCGA344-351 and bCGA 348-358 are antibacterial at 20 µM.


**Association of Electronegative LDL with Macrophage Foam Cell Formation and CD11c Expression in Rheumatoid Arthritis Patients**

**Ching-Kun Chang 1,2, Po-Ku Chen 1,2,3, Joung-Liang Lan 2,3,4, Shih-Hsin Chang 2,3,5, Tsu-Yi Hsieh 6, Pei-Jyuan Liao 1,2, Chu-Huang Chen 7,8 and Der-Yuan Chen 1,2,3,5,\***

Received: 20 July 2020; Accepted: 14 August 2020; Published: 16 August 2020

**Abstract:** L5, the most negatively charged subfraction of low-density lipoprotein (LDL), is implicated in atherogenesis, but the pathogenic association is relatively unexplored in patients with rheumatoid arthritis (RA). We examined the role of L5 LDL in macrophage foam cell formation and the association of L5 with CD11c expression in THP-1 cells and RA patients. Using quantitative real-time PCR, we determined mRNA expression levels of *ITGAX*, the gene for CD11c, a marker associated with vascular plaque formation and M1 macrophages in atherogenesis, in 93 RA patients. We also examined CD11c expression on THP-1 cells treated with L5 by flow cytometry analysis and the plasma levels of inflammatory mediators using a magnetic bead array. We found a dose-dependent upregulation of foam cell formation of macrophages after L5 treatment (mean ± SEM, 12.05 ± 2.35% in L5 (10 μg/mL); 50.13 ± 3.9% in L5 (25 μg/mL); 90.69 ± 1.82% in L5 (50 μg/mL), *p* < 0.01). Significantly higher levels of CD11c expression were observed in 30 patients with a high percentage of L5 in LDL (L5%) (0.0752 ± 0.0139-fold) compared to 63 patients with normal L5% (0.0446 ± 0.0054-fold, *p* < 0.05). CD11c expression levels were increased in the L5-treated group (30.00 ± 3.13% in L5 (10 μg/mL); 41.46 ± 2.77% in L5 (50 μg/mL), *p* < 0.05) and were positively correlated with plasma levels of interleukin (IL)-6 and IL-8. L5 augmented the expression of IL-6, IL-8, and tumor necrosis factor-α (TNF-α) on monocytes and macrophages. Our findings suggest that L5 may promote atherogenesis by augmenting macrophage foam cell formation, upregulating CD11c expression, and enhancing the expression levels of atherosclerosis-related mediators.

**Keywords:** L5; macrophage foam cell; CD11c expression; atherosclerosis; rheumatoid arthritis (RA)


2. Governing equations

The equation of continuity and momentum of MHD flow passing through porous space is given by [7]

$$\nabla \cdot V = 0,\\ \rho \left(\frac{d\mathbf{V}}{dt}\right) = d\dot{v}\mathbf{r}\mathbf{T} + \mathbf{J} \times \mathbf{B} + \mathbf{R},\tag{1}$$

where velocity is represented by V, density by ρ, Cauchy stress tensor by T, magnetic body force by J � B, current density by J, magnetic field by B, and Darcy's resistance in the porous medium by R.

For an incompressible and unsteady Jeffrey fluid the Cauchy stress tensor is defined as [9]

$$\mathbf{T} = -p\mathbf{I} + \mathbf{S}, \ \mathbf{S} = \frac{\mu}{1+\lambda} \left[ \mathbf{A} + \Theta \left( \frac{\partial \mathbf{A}}{\partial \mathbf{t}} + (\mathbf{V} \cdot \nabla) \mathbf{A} \right) \right],\tag{2}$$

where S and pI represents the extra stress tensor and the indeterminate spherical stress, the dynamic viscosity is denoted by μ, A= L + LT is the first Rivlin-Ericksen tensor, L is the velocity gradient, λ is relaxation time and θ is retardation time. The Lorentz force due to magnetic field is

$$\mathbf{J} \times \mathbf{B} = -\sigma \beta\_o^2 \mathbf{V},\tag{3}$$

where σ represents electrical conductivity and βo the strength of magnetic field. For the Jeffrey fluid the Darcy's resistance satisfies the following equation

$$\mathbf{R} = -\frac{\mu \phi}{\kappa (1 + \lambda)} \left( 1 + \theta \frac{\partial}{\partial t} \right) \mathbf{V},\tag{4}$$

where κ(>0) and ϕ(0 < ϕ < 1) are the permeability and the porosity of the porous medium.

In the following problem we consider a velocity field and extra stress of the form

$$\mathbf{V} = (0, 0, w(\mathbf{x}, y, t)), \mathbf{S} = \mathbf{S}(\mathbf{x}, y, t) \tag{5}$$

where w is the velocity in the z-direction. The continuity equation for such flows is automatically satisfied. Also, at t = 0, the fluid being at rest is given by

$$\mathbf{S}(\mathbf{x}, y, \mathbf{0}) = \mathbf{0},\tag{6}$$

therefore from Eqs. (2), (5) and (6), it results that Sxx = Syy = Syz = Szz = 0 and the relevant equations

$$
\tau\_1 = \frac{\mu}{(1+\lambda)} \left( 1 + \theta \frac{\partial}{\partial t} \right) \partial\_x w(\mathbf{x}, y, t), \quad \tau\_2 = \frac{\mu}{(1+\lambda)} \left( 1 + \theta \frac{\partial}{\partial t} \right) \partial\_y w(\mathbf{x}, y, t), \tag{7}
$$

where τ1 = Sxy and τ2 = Sxz are the tangential stresses. In the absence of pressure gradient in the flow direction, the governing equation leads to

$$\mathbf{u}(1+\lambda)\partial\_t \mathbf{w}(\mathbf{x}, y, t) = \nu \left( 1 + \theta \frac{\partial}{\partial t} \right) \left( \partial\_x^2 + \partial\_y^2 \right) \mathbf{w}(\mathbf{x}, y, t) - \nu \mathbf{K} \left( 1 + \theta \frac{\partial}{\partial t} \right) \mathbf{w}(\mathbf{x}, y, t) - H(1+\lambda)\mathbf{w}(\mathbf{x}, y, t), \tag{8}$$

where H ¼ σB2 0 ρ is the magnetic parameter, K ¼ ϕ κ is the porosity parameter and ν = μ/ρ is the kinematic viscosity.


*Edited by Habtamu Alem*

In the face of unprecedented challenges posed by the global COVID-19 pandemic, *Resilience and Realities - Exploring Pandemic Effects, Governance Challenges, and Economic Insights* offers a comprehensive exploration of the intricate interplay between public policies and uncertain times. This enlightening volume presents a thought-provoking collection of chapters that dissect the illusions, opportunities, and complexities surrounding public policies during crises. Spanning two captivating sections, this book embarks on a journey through the labyrinth of pandemic-related public policies, offering fresh insights into the ever-evolving landscape of economic responses and the resilient spirit of entrepreneurship. In the first section, "Illusions of Public Policies Amidst a Pandemic", readers are invited to dissect the enigma of navigating uncertainties in crisis management. Delve into the gendered impact of the pandemic on academic women, explore the untapped opportunities within the pandemic's wake, and scrutinize the efficacy of governmental economic strategies. The second section, "Public Policies towards Investment, Technology, and Efficiency", provides an insightful investigation into the impact of investment in human capital on economic growth. Review the intricacies of financial management within the public sector and gain an understanding of the multifaceted dimensions of technology, investment, and efficiency in public policies.With each chapter, esteemed authors contribute their expertise to illuminate the complexities of these critical topics. *Resilience and Realities - Exploring Pandemic Effects, Governance Challenges, and Economic Insights* is a vital resource for policymakers, economists, academics, and curious minds seeking a deeper understanding of the intricate nexus between public policies and uncertain times. Explore the wealth of knowledge and insights within these pages and embark on a journey toward a more informed and resilient future. Discover illuminating perspectives, engage in thought-provoking analyses, and embrace the ever-changing landscape of public economics. This book stands as a beacon of knowledge and enlightenment in an era of unprecedented challenges.

> *Usha Iyer-Raniga, Sustainable Development Series Editor*

> > ISBN 978-1-83880-770-2

Resilience and Realities - Exploring Pandemic Effects, Governance Challenges, and Economic Insights

ISSN 2753-6580

Published in London, UK © 2024 IntechOpen © Andre2013 / iStock

IntechOpen Series Sustainable Development, Volume 11


**Author details**

Tetsuya Endo1 \* and Mari Kobayashi2

1 School of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Hokkaido, Japan

2 Faculty of Bioindustry, Tokyo University of Agriculture, Abashiri, Hokkaido, Japan

\*Address all correspondence to<EMAIL_ADDRESS>
© 2022 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, provided the original work is properly cited.