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For decades, lactic acid bacteria has been isolated and selected to be used as starter cultures in meat fermentation for standardization and management of quality of dry‐fermented sausage which constitute a considerable challenge. The aim of this study was to evaluate the effect of Lactobacillus sakei strains, isolated from different origins, on qualities of dry‐fermented sausages. These last, manufactured with different combinations of starter cultures (L. sakei + Staphylococcus xylosus), were ripened, using the same raw materials and conditions, for 45 days. Samples were collected during this period, and microbiological, physicochemical, fatty acid profile, and sensorial analyses determined. Lactic acid bacteria were the dominant flora during ripening. A desirable PUFA/SFA ratio, corresponding to 1:1.7 (0.6), was detected after 24 days of maturation in sausages inoculated by L. sakei BMG 95 and S. xylosus. Sensory analysis showed that fermented sausages manufactured with L. sakei and S. xylosus had a more desirable odor, flavor, and texture and consequently were preferred overall. In particular, sensory panellists preferred sausages produced with either L. sakei 23K or L. sakei BMG 95 when compared to fermented sausage produced with a commercial starter or no starter at all.

A total of 15 samples of thalassotherapy products, distributed in Tunisia in their intact and final state of production, was analyzed to determine their microbiological safety status. The result shows the absence of pathogenic bacteria (Staphylococcus aureus, Candida albicans, Salmonella, Pseudomonas aeruginosa and coliforms). The incidence of contamination by Gram-positive Bacilli (mesophelic bacteria, aerobic and anaerobic spore forming bacteria, anaerobic sulphite-reducing bacteria) was found to be higher in products composed by mud and extract of alga. The biochemical and molecular identification of the major contaminant show that Bacilli were the most covered from 75% of the thalassotherapy products. Mineral analysis (organic matter, Fe, Mg, Ca, Na d K, Al, Si and Ti) shows strong composition on Aluminum and Silica. Cytotoxicity study of six thalassotherapy products and three essential oil extracts (Menthol, Clove and Eucalyptus) did not show any cytotoxic effect. Furthermore, antibacterial acitivity of 5 essentila oils, against 30 isolates of the genus Bacillus and 10 reference strains, has been characterized showing an interesting bactericidal potential of the extract of menthol and Eucalyptus.

SummaryCoronavirus disease 2019 (COVID-19) expresses a wide spectrum of disease severity. We investigated the profile of IgG and IgG subclass antibody responses to SARS– CoV-2 in Tunisian patients with COVID-19 according to disease severity (86 patients with severe disease and 63 with mild to moderate disease). Two in house developed ELISA with excellent performance were used to test for antibodies to the nucleocapsid (N) protein and the receptor-binding domain of the spike antigen (S-RBD) of SARS-CoV-2. IgG, IgG1 and IgG3 antibodies were significantly higher in patients with severe disease compared to non-severe disease. Antibodies to S-RBD or the N protein were dominated by IgG1 and IgG3 or IgG1/IgG3 and IgG2 subclasses respectively. In patients with severe disease, IgG antibodies ‘ appearance to S-RBD was delayed compared to the N protein. IgG subclass imbalance may reflect the pathophysiology of COVID-19 and may herald disease aggravation. This study brings information on the immune responses to SARS-CoV-2 in North African patients and completes the picture drawn on COVID-19 in different African populations and worldwide.

Hepcidin a 25-amino-acid and highly disulfide bonded hormone, is the central regulator of iron homeostasis. In this chapter we propose ferritin as a peptide carrier to promote the association of the hybrid hepcidin/ferritin nanoparticle with a particular cell or tissue for therapeutic or diagnostic use. Indeed, human ferritin H-chain fused directly (on its 5’end) with camel mature hepcidin was cloned into the pASK-43 plus vector and expressed using BL21 (DE3) pLys E. coli strain. The transformed E.coli produced efficiently hepcidin-ferritin construct (hepcH), consisting of 213 amino acids with a molecular weight of 24 KDa. The recovered product is a ferritin exposing hepcidin on outer surface. The hepcH monomer was characterized by immunoblotting using a monoclonal antibody specific for human ferritin and a polyclonal antibody specific for hepcidin-25. The results were also confirmed by MALDI-TOF mass spectrometry. The recombinant native human ferritin and the commercial human hepcidin-25 were used as controls in this experiment. The assembly of hepcH, as an heteropolymer molecule, was performed in presence of denatured human ferritin-H and -L chains. After cysteine oxidation of the recombinant nanoparticles, cellular binding assays were performed on mammalian cells such as mouse monocyte–macrophage cell line J774, HepG2 and COS7.

The use of starter cultures in the industrial manufacture of meat products has become common practice. We have investigated the use of Lactobacillus sakei F, isolated from a commercial starter culture, and Lactobacillus sakei 23K, from a French sausage, alone or together with Staphyloccus xylosus, as starter cultures, during the industrial manufacture of Tunisian dry‐fermented sausages. Samples from the same raw matter were collected after up to 45 days of maturation and were subjected to microbiological, physicochemical, nutritional, and sensory analyses. Furthermore, we demonstrated that the use of L. sakei F or L. sakei 23K as starter strains produced a fermented sausage with similar or even better quality than sausages fermented with L. sakei 23K + S. xylosus or the commercial starter. Our study found that L. sakei 23K could have a great potential for use as a starter culture during the commercial production of dry‐fermented sausage. NOVELTY IMPACT STATEMENT: Lactobacillus sakei F and 23K strains can be used alone or in association with Staphylococcus xylosus as starter cultures for the manufacture of Tunisian dry‐fermented sausages. Sausages produced with L. sakei F and 23K strains display better nutritional and sensory profiles, compared with the non‐inoculated controls, as well as sausages inoculated with the commercial starter culture.

Hepatitis E virus (HEV) is a nonenveloped virus causing an emerging zoonotic disease posing a severe threat to the public health in the world, especially to pregnant women. In this study, a truncated form (aa 368-606) of the open reading frame 2 of the capsid protein (tORF2-HEV), a major structural protein of HEV, was expressed in Escherichia coli. This work characterizes for the first time, the fused Glutathione-S-Transferase-tagged tORF2 (GST-tORF2) and tORF2-HEV forms in E. coli. The fusion protein was purified by affinity chromatography with a purity higher than 90% and to yield about 27% after thrombin digestion. The purified GST-tORF2 protein was then characterized by western blot, using anti-GST antibodies, and CD spectroscopy. The GST-tORF2 and tORF2-HEV proteins were shown to be efficient to develop an ELISA test to detect anti-HEV IgG in mice sera immunized with a recombinant full length ORF2 protein. Sera showed a significant increase of the absorbance signal at 450 nm, in plate wells coated with a quantity of 0.5, 1 and 2 µg of proteins. ELISA plates coated with the purified GST-tORF2 and tORF2-HEV showed similar response when compared to the HEV ELISA where total insect cell lysate, infected with the recombinant baculovirus expressing full ORF2, was used as positive control.

Hepatitis E virus (HEV) is a nonenveloped virus causing an emerging zoonotic disease posing a severe threat to the public health in the world, especially to pregnant women. In this study, a truncated form (aa 368-606) of the open reading frame 2 of the capsid protein (

In this paper, using the technique of measure of weak noncompactness and by removing the convexity hypothesis, we prove some fixed set results for the sum and the product of three multivalued mappi...

For decades, lactic acid bacteria has been isolated and selected to be used as starter cultures in meat fermentation for standardization and management of quality of dry‐fermented sausage which constitute a considerable challenge. The aim of this study was to evaluate the effect of Lactobacillus sakei strains, isolated from different origins, on qualities of dry‐fermented sausages. These last, manufactured with different combinations of starter cultures (L. sakei + Staphylococcus xylosus), were ripened, using the same raw materials and conditions, for 45 days. Samples were collected during this period, and microbiological, physicochemical, fatty acid profile, and sensorial analyses determined. Lactic acid bacteria were the dominant flora during ripening. A desirable PUFA/SFA ratio, corresponding to 1:1.7 (0.6), was detected after 24 days of maturation in sausages inoculated by L. sakei BMG 95 and S. xylosus. Sensory analysis showed that fermented sausages manufactured with L. sakei and S. xylosus had a more desirable odor, flavor, and texture and consequently were preferred overall. In particular, sensory panellists preferred sausages produced with either L. sakei 23K or L. sakei BMG 95 when compared to fermented sausage produced with a commercial starter or no starter at all., Dry‐fermented sausage samples inoculated with Lactobacillus sakei BMG 95 showed best physicochemical, nutritional, and sensory satisfactory results in comparison with the commercial starter. This strain, isolated from anchovies, was able to produce a fermented sausage with similar/or better quality as sausages fermented with the commercial starter.

Ferritin is a molecule with enormous potentiality in biotechnology that have been already used to encapsulate molecules, as contrast in magnetic resonance imaging and to carry epitopes. We proposed to use it to carry another key protein of iron metabolism, hepcidin that is a small hormone peptide that control systemic iron homeostasis. In this work, we purified the previously produced camel hepcidin and human H-ferritin heteropolymer (HepcH-FTH) and to monitor its binding capability toward J744 cell line in presence or absence of ferric ammonium citrate. Fused camel hepcidin and human H-ferritin monomer (HepcH) as well as the assembled HepcH-FTH heteropolymer (ratio 1:5) was easily purified by a one-step purification using size exclusion chromatography. SDS-PAGE electrophoresis of HepcH, purified from soluble and insoluble fractions, showed a single band of 24 kDa with an estimated purity of at least 90%. The purification yields of HepcH from the soluble and insoluble fractions was, respectively, of about 6.80 and 2 mg/L of bacterial culture. Time curse cellular binding assays of HepcH-FTH revealed its great potential to bind the J774 cells after 15 min of incubation. Furthermore, HepcH-FTH was able to degrade ferroportin, the unique hepcidin receptor, even after 30 min of incubation with J774 cells treated with 100 µM ferric ammonium citrate. In conclusion, we proposed ferritin as a peptide carrier to promote the association of the hybrid HepcH-FTH nanoparticle with a particular type of cell for therapeutic or diagnostic.

We introduce the concept of Meir-Keeler condensing operator in a Banach space via an arbitrary measure of weak noncompactness. We prove some generalizations of Darbo's fixed point theorem by considering a measure of weak noncompactness which not necessary has the maximum property. We prove some coupled fixed point theorems and we apply them in order to establish the existence of weak solutions for a system of functional integral equations of Volterra type.

In this paper, we introduce a new class of multivalued mappings of the form \({\frac{I-C}{A}}\), where \({A}\) and \({C}\) are multivalued mappings acting on Banach algebras. Using this concept, we present some fixed point theorems in Banach algebras satisfying a certain sequential condition (\({P}\)) in the weak topology setting. Our main tool is the axiomatic measure of weak noncompactness. Our results generalize, extend and improve wellknown results on the subject. An application to illustrate our theory is included.

Hepcidin, belonging to the β-defensin family, was isolated for the first time from plasma and human urine. It is a cationic peptide, rich in cysteine bound with four disulfide bridges, which plays a major role in innate immunity and iron homeostasis. Some vertebrate species have multiple hepcidin homolog genes and each contains only one copy that functions as an iron regulator except hepcidin sequences in the pigeon (Columba livia). The aim of this chapter is to investigate the molecular evolution of several hepcidin gene from searches of the literature and public genomic databases from 17 different species, all among the vertebrates.

Hepcidin, belonging to the β-defensin family, was isolated for the first time from plasma and human urine. It is a cationic peptide, rich in cysteine bound with four disulfide bridges, which plays a major role in innate immunity and iron homeostasis. Some vertebrate species have multiple hepcidin homolog genes and each contains only one copy that functions as an iron regulator except hepcidin sequences in the pigeon (Columba livia). The aim of this chapter is to investigate the molecular evolution of several hepcidin gene from searches of the literature and public genomic databases from 17 different species, all among the vertebrates.

Background Recent biotechnological advancements have allowed for the adoption of Lactococcus lactis, a typical component of starter cultures used in food industry, as the host for the production of food-grade recombinant targets. Among several advantages, L. lactis has the important feature of growing on lactose, the main carbohydrate in milk and a majoritarian component of dairy wastes, such as cheese whey. Results We have used recombinant L. lactis NZ9000 carrying the nisin inducible pNZ8148 vector to produce MNEI, a small sweet protein derived from monellin, with potential for food industry applications as a high intensity sweetener. We have been able to sustain this production using a medium based on the cheese whey from the production of ricotta cheese, with minimal pre-treatment of the waste. As a proof of concept, we have also tested these conditions for the production of MMP-9, a protein that had been previously successfully obtained from L. lactis cultures in standard growth conditions. Conclusions Other than presenting a new system for the recombinant production of MNEI, more compliant with its potential applications in food industry, our results introduce a strategy to valorize dairy effluents through the synthesis of high added value recombinant proteins. Interestingly, the possibility of using this whey-derived medium relied greatly on the choice of the appropriate codon usage for the target gene. In fact, when a gene optimized for L. lactis was used, the production of MNEI proceeded with good yields. On the other hand, when an E. coli optimized gene was employed, protein synthesis was greatly reduced, to the point of being completely abated in the cheese whey-based medium. The production of MMP-9 was comparable to what observed in the reference conditions. Electronic supplementary material The online version of this article (10.1186/s12934-018-0974-z) contains supplementary material, which is available to authorized users.

Enzymatic saccharification of lignocellulosic biomass has been widely studied. Mainly endoglucanases were found to be a prerequisite for the quick initial biomass liquefaction. In the present study, Pichia pastoris was used as a host for the heterologous expression of a Sclerotinia sclerotiorum GH45 endoglucanase, Endo2. The recombinant plasmid pPICZαA was used to transform Pichia pastoris. Pichia culture supernatants expressing the recombinant Endo2 (rEndo2) were used for the purification and biochemical characterization of this enzyme. Therefore, rEndo2 was purified 6.7 fold to homogeneity with 34% yield and gave 19U/mg specific activity. It also showed maximum activity at pH 7.0 and 60°C (against pH 5.0 and 50°C for the native enzyme) and was thermostable at relatively high temperatures. Furthermore, rEndo2 retained its activity in a wide pH range (from 5 to 8). Besides, the recombinant endoglucanase was produced as an active 47kDa enzyme. This molecular weight differs from the one of the native enzyme (34kDa), which suggested a potential glycosylation of the recombinant enzyme. Moreover, rEndo2 was able to produce fermentable sugars after enzymatic assay on various cellulosic substrates with an interesting yield. Therefore, all these features offer prospects for large-scale production and industrial application of the recombinant endoglucanase.

Hepcidin is a cysteine-rich peptide widely characterized in immunological processes and antimicrobial activity in several vertebrate species. Obviously, this hormone plays a central role in the regulation of systemic iron homeostasis. However, its role in camelids' immune response and whether it is involved in antibacterial immunity have not yet been proven. In this study, we characterized the Arabian camel hepcidin nucleotide sequence with an open reading frame of 252 bp encoding an 83-amino acid preprohepcidin peptide. Eight cysteine key residues conserved in all mammalian hepcidin sequences were identified. The model structure analysis of hepcidin-25 peptide showed a high homology structure and sequence identity to the human hepcidin. Two different hepcidin-25 analogs manually synthesized by SPPS shared significant cytotoxic capacity toward the Gram-negative bacterium Escherichia coli American Type Culture Collection (ATCC) 8739 as well as the Gram-positive bacteria Bacillus subtilis ATCC 11779 and Staphylococcus aureus ATCC 6538 in vitro. The three disulfide bridges hepcidin analog demonstrated bactericidal activity, against B. subtilis ATCC 11779 and S. aureus ATCC 6538 strains, at the concentration of 15 μM (50 µg/ml) or above at pH 6.2. This result correlates with the revealed structural features suggesting that camel hepcidin is proposed to be involved in antibacterial process of innate immune response. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.

This article has been accepted for publication in Protein Engineering design and Selection Published by Oxford University Press., Hepcidin is a liver-synthesized hormone that plays a central role in the regulation of systemic iron homeostasis. To produce a new tool for its functional properties the cDNA coding for camel hepcidin-25 was cloned at the 5’end of human FTH sequence into the pASK-IBA43plus vector for expression in Escherichia coli. The recombinant fusion hepcidin–ferritin-H subunit was isolated as an insoluble iron-containing protein. When alone it did not refold in a 24-mer ferritin molecule, but it did when renatured together with H- or L-ferritin chains. We obtained stable ferritin shells exposing about 4 hepcidin peptides per 24-mer shell. The molecules were then reduced and re-oxidized in a controlled manner to allow the formation of the proper hepcidin disulfide bridges. The functionality of the exposed hepcidin was confirmed by its ability to specifically bind the mouse macrophage cell line J774 that express ferroportin and to promote ferroportin degradation. This chimeric protein may be useful for studying the hepcidin–ferroportin interaction in cells and also as drug-delivery agent., This work is partially financed by the Laboratory of Protein Engineering and Bioactive Molecules (LIP-MB) and the Doctoral School of the National Institute of Applied Sciences and Technology (INSAT-Tunis) – University of Carthage.

Hepcidin, a liver-expressed antimicrobial peptide, has been demonstrated to act as an iron regulatory hormone as well as to exert a wide spectrum of antimicrobial activity. The aim of this work was the expression, as secreted peptide, purification, and characterization of a new recombinant polyHis-tagged camel hepcidin (HepcD-His) in yeast Pichia pastoris. The use of this eukaryotic expression system, for the production of HepcD-His, having 6 histidine residues at its C terminus, was simpler and more efficient compared with the use of the prokaryotic system Escherichia coli. Indeed, a single purification step was required to isolate the soluble hepcidin with purity estimated more that 94% and a yield of 2.8 against 0.2 mg/L for the E coli system. Matrix-assisted laser desorption/ionization time-of-flight (TOF)/TOF mass spectrometry of the purified HepcD-His showed 2 major peaks at m/z 4524.64 and 4634.56 corresponding to camel hepcidin with 39 and 40 amino acids. Evaluation of disulfide bond connectivity with the Ellman method showed an absence of free thiol groups, testifying that the 8 cysteine residues in the peptide are displayed, forming 4 disulfide bridges. Circular dichroism spectroscopy showed that camel hepcidin structure was significantly modified at high temperature of 90°C and returns to its original structure when incubation temperature drops back to 20°C. Interestingly, this peptide showed also a greater bactericidal activity, at low concentration of 9.5μM, against E coli, than the synthetic analog DH3. Thus, the production, at a large scale, of the recombinant camel hepcidin, HepcD-His, may be helpful for future therapeutic applications including bacterial infection diseases.

Resume Soit q : E → P une algebroide de Courant. On se propose de munir le fibre tangent T q : T E → T P d'une structure d'algebroide de Courant relevee de celle de q : E → P . En particulier, dans le cas ou E = A ⊕ A ∗ le double d'une bigebroide de Lie ( A , A ∗ ) , l'algebroide de Courant tangente TE est le double de la bigebroide de Lie tangente ( T A , ( T A ) ∗ ) . Les structures de Dirac de l'algebroide tangente TE sont determinees a partir des sous-fibres integrables de E. Pour citer cet article : M. Boumaiza, N. Zaalani, C. R. Acad. Sci. Paris, Ser. I 347 (2009).

International audience; Hepcidin, a 25-amino-acid and highly disulfide bonded antimicrobial peptide, is the central regulator of iron homeostasis. This hormone is expressed in response to iron and inflammation and interacts with ferroportinl (FPN1), the only known iron exporter in vertebrates, inducing its internalization and degradation. Thus, the export of iron from cells to plasma will be significantly diminished. Thereby, hepcidin has become the target of intense research studies due to its profound biomedical significance. This study describes the functional expression of recombinant camel hepcidin in Escherichia coli. Biologically active recombinant camel hepcidin was obtained thanks to the production of a hepcidin-thioredoxin fusion protein (TRX-HepcD) and a purified camel hepcidin, with an extra methionine at the N-terminus, was obtained after enterokinase cleavage of the fusion protein. Presence of the four disulfide bridges was verified using MALDI-ToF spectrometry. The recombinant camel hepcidin was compared to related synthetic bioactive peptides, including human hepcidin, and was found equally able to promote ferroportin degradation of mouse macrophages. Furthermore, camel hepcidins exhibits a high capacity to inhibit the growth of Leishmania major promastigotes. These results proved that production of functional camel hepcidin can be achieved in E. coli, this is a major interest for the production of cysteine rich peptides or proteins that can be purified under their functional form without the need of a refolding process. (C) 2015 Elsevier Inc. All rights reserved.

Hepcidin is a cysteine-rich peptide widely characterized in immunological processes and antimicrobial activity in several vertebrate species. Obviously, this hormone plays a central role in the regulation of systemic iron homeostasis. However, its role in camelids' immune response and whether it is involved in antibacterial immunity have not yet been proven. In this study, we characterized the Arabian camel hepcidin nucleotide sequence with an open reading frame of 252 bp encoding an 83-amino acid preprohepcidin peptide. Eight cysteine key residues conserved in all mammalian hepcidin sequences were identified. The model structure analysis of hepcidin-25 peptide showed a high homology structure and sequence identity to the human hepcidin. Two different hepcidin-25 analogs manually synthesized by SPPS shared significant cytotoxic capacity toward the Gram-negative bacterium Escherichia coli American Type Culture Collection (ATCC) 8739 as well as the Gram-positive bacteria Bacillus subtilis ATCC 11779 and Staphylococcus aureus ATCC 6538 in vitro. The three disulfide bridges hepcidin analog demonstrated bactericidal activity, against B. subtilis ATCC 11779 and S. aureus ATCC 6538 strains, at the concentration of 15 μM (50 µg/ml) or above at pH 6.2. This result correlates with the revealed structural features suggesting that camel hepcidin is proposed to be involved in antibacterial process of innate immune response.

Hepcidin, a liver-expressed antimicrobial peptide, has been demonstrated to act as an iron regulatory hormone as well as to exert a wide spectrum of antimicrobial activity. The aim of this work was the expression, as secreted peptide, purification, and characterization of a new recombinant polyHis-tagged camel hepcidin (HepcD-His) in yeast Pichia pastoris. The use of this eukaryotic expression system, for the production of HepcD-His, having 6 histidine residues at its C terminus, was simpler and more efficient compared with the use of the prokaryotic system Escherichia coli. Indeed, a single purification step was required to isolate the soluble hepcidin with purity estimated more that 94% and a yield of 2.8 against 0.2 mg/L for the E coli system. Matrix-assisted laser desorption/ionization time-of-flight (TOF)/TOF mass spectrometry of the purified HepcD-His showed 2 major peaks at m/z 4524.64 and 4634.56 corresponding to camel hepcidin with 39 and 40 amino acids. Evaluation of disulfide bond connectivity with the Ellman method showed an absence of free thiol groups, testifying that the 8 cysteine residues in the peptide are displayed, forming 4 disulfide bridges. Circular dichroism spectroscopy showed that camel hepcidin structure was significantly modified at high temperature of 90°C and returns to its original structure when incubation temperature drops back to 20°C. Interestingly, this peptide showed also a greater bactericidal activity, at low concentration of 9.5μM, against E coli, than the synthetic analog DH3. Thus, the production, at a large scale, of the recombinant camel hepcidin, HepcD-His, may be helpful for future therapeutic applications including bacterial infection diseases.

We show in this paper that the tangent bundle $TG$, of a Poisson-Lie group $G$ has a Poisson-Lie group structure given by the canonical lifting of that of $G$. We determine the dual group of $TG$, its Lie bialgebra and its double Lie algebra. ¶ We also show that any Poisson action of $G$ on a Poisson manifold $P$ is lifted on a Poisson action of $TG$ on the tanget bundle $TP$.