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Article ; Online: Leishmania infantum transfected with toxic plasmid induces protection in mice infected with wild type L. infantum or L. amazonensis.

Zorgi, Nahiara Esteves / Arruda, Leonardo V / Paladine, Izadora / Roque, Guilherme A S / Araújo, Thalita F / Brocchi, Marcelo / Barral, Manoel / Sanchiz, África / Requena, José María / Abánades, Daniel R / Giorgio, Selma

Molecular immunology

2020 Volume 127, Page(s) 95–106

Abstract: ... in the intracellular form of L. infantum, as demonstrated by fluorescence microscopy, flow cytometry and ...

Abstract	Leishmania infantum infection may cause visceral leishmaniasis (VL), a fatal disease having worldwide distribution, that may be silent or asymptomatic. The latter indicates that immunity is naturally developed in some individuals, and, therefore, a vaccine against VL would be possible. Molecular mechanisms of gene expression are being understood in Leishmania, and this knowledge may be useful for vaccine development. The aim of this study was developing an attenuated strain by regulating the expression of toxic proteins in a stage specific manner. For that purpose, the 3' UTR of an amastin gene, known by its increased expression in the amastigote phase, was selected for direct the expression of exogenous proteins. This construct (pFL-AMA), firstly, was proved effective for the expression of mCherry specifically in the intracellular form of L. infantum, as demonstrated by fluorescence microscopy, flow cytometry and Western blotting. Afterwards, mCherry coding sequence was replaced, in the pFL-AMA plasmid, by either egg avidin or the active form of bovine trypsin. Viability of transfected parasites was evaluated in promastigote axenic cultures and in in vitro infection of macrophages. Both lines of transfected parasites showed a limited capacity to multiply inside macrophages. BALB/c mice were inoculated intraperitoneally (i.p.) with a single dose consisting of 2 × 10
MeSH term(s)	Animals ; Female ; Gene Expression Regulation ; Humans ; Immunity, Cellular ; Immunity, Humoral ; Immunization ; Leishmania/pathogenicity ; Leishmania/physiology ; Leishmania infantum/growth & development ; Leishmania infantum/pathogenicity ; Leishmania infantum/physiology ; Leishmaniasis/immunology ; Leishmaniasis/parasitology ; Leishmaniasis/prevention & control ; Life Cycle Stages ; Mice, Inbred BALB C ; Parasites/metabolism ; Parasites/pathogenicity ; Plasmids/metabolism ; Protozoan Proteins/metabolism ; Toxins, Biological/metabolism ; Transfection ; Virulence
Chemical Substances	Protozoan Proteins ; Toxins, Biological
Language	English
Publishing date	2020-09-17
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	424427-8
ISSN	1872-9142 ; 0161-5890
ISSN (online)	1872-9142
ISSN	0161-5890
DOI	10.1016/j.molimm.2020.08.006
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Article ; Online: Inoculation of the Leishmania infantum HSP70-II Null Mutant Induces Long-Term Protection against L. amazonensis Infection in BALB/c Mice

Manuel Soto / Laura Ramírez / José Carlos Solana / Emma C. L. Cook / Elena Hernández-García / José María Requena / Salvador Iborra

Microorganisms, Vol 9, Iss 2, p

2021 Volume 363

Abstract: ... BALB/c mice are highly susceptible to L. amazonensis challenge due to their inability to mount parasite ... of the LiΔHSP70-II genetically-modified attenuated L. infantum line in preventing cutaneous leishmaniasis in mice ... challenged with L. amazonensis virulent parasites. In previous studies, this live attenuated vaccine has ...

Abstract	Leishmania amazonensis parasites are etiological agents of cutaneous leishmaniasis in the New World. BALB/c mice are highly susceptible to L. amazonensis challenge due to their inability to mount parasite-dependent IFN-γ-mediated responses. Here, we analyzed the capacity of a single administration of the LiΔHSP70-II genetically-modified attenuated L. infantum line in preventing cutaneous leishmaniasis in mice challenged with L. amazonensis virulent parasites. In previous studies, this live attenuated vaccine has demonstrated to induce long-protection against murine leishmaniasis due to Old World Leishmania species. Vaccinated mice showed a reduction in the disease evolution due to L. amazonensis challenge, namely reduction in cutaneous lesions and parasite burdens. In contrast to control animals, after the challenge, protected mice showed anti- Leishmania IgG2a circulating antibodies accompanied to the induction of Leishmania -driven specific IFN-γ systemic response. An analysis performed in the lymph node draining the site of infection revealed an increase of the parasite-specific IFN-ϒ production by CD4 + and CD8 + T cells and a decrease in the secretion of IL-10 against leishmanial antigens. Since the immunity caused by the inoculation of this live vaccine generates protection against different forms of murine leishmaniasis, we postulate LiΔHSP70-II as a candidate for the development of human vaccines.
Keywords	Leishmania amazonensis ; live vaccines ; attenuated parasites ; murine leishmaniasis ; BALB/c mice ; IFN-γ ; Biology (General) ; QH301-705.5
Language	English
Publishing date	2021-02-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Inoculation of the Leishmania infantum HSP70-II Null Mutant Induces Long-Term Protection against L. amazonensis Infection in BALB/c Mice

Soto, Manuel / Ramírez, Laura / Solana, José Carlos / Cook, Emma C. L. / Hernández-García, Elena / Requena, José María / Iborra, Salvador

Microorganisms. 2021 Feb. 12, v. 9, no. 2

2021

Abstract	Leishmania amazonensis parasites are etiological agents of cutaneous leishmaniasis in the New World. BALB/c mice are highly susceptible to L. amazonensis challenge due to their inability to mount parasite-dependent IFN-γ-mediated responses. Here, we analyzed the capacity of a single administration of the LiΔHSP70-II genetically-modified attenuated L. infantum line in preventing cutaneous leishmaniasis in mice challenged with L. amazonensis virulent parasites. In previous studies, this live attenuated vaccine has demonstrated to induce long-protection against murine leishmaniasis due to Old World Leishmania species. Vaccinated mice showed a reduction in the disease evolution due to L. amazonensis challenge, namely reduction in cutaneous lesions and parasite burdens. In contrast to control animals, after the challenge, protected mice showed anti-Leishmania IgG2a circulating antibodies accompanied to the induction of Leishmania-driven specific IFN-γ systemic response. An analysis performed in the lymph node draining the site of infection revealed an increase of the parasite-specific IFN-ϒ production by CD4⁺ and CD8⁺ T cells and a decrease in the secretion of IL-10 against leishmanial antigens. Since the immunity caused by the inoculation of this live vaccine generates protection against different forms of murine leishmaniasis, we postulate LiΔHSP70-II as a candidate for the development of human vaccines.
Keywords	Leishmania amazonensis ; Leishmania infantum ; cutaneous leishmaniasis ; disease progression ; humans ; immunity ; immunoglobulin G ; interleukin-10 ; live vaccines ; lymph nodes ; mice ; mutants ; parasites ; secretion ; virulence
Language	English
Dates of publication	2021-0212
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms9020363
Database	NAL-Catalogue (AGRICOLA)

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Article: Pyrometric-Based Melt Pool Monitoring Study of CuCr1Zr Processed Using L-PBF.

Artzt, Katia / Siggel, Martin / Kleinert, Jan / Riccius, Joerg / Requena, Guillermo / Haubrich, Jan

Materials (Basel, Switzerland)

2020 Volume 13, Issue 20

Abstract: The potential of in situ melt pool monitoring (MPM) for parameter development and furthering the process understanding in Laser Powder Bed Fusion (LPBF) of CuCr1Zr was investigated. Commercial MPM systems are currently being developed as a quality ... ...

Abstract	The potential of in situ melt pool monitoring (MPM) for parameter development and furthering the process understanding in Laser Powder Bed Fusion (LPBF) of CuCr1Zr was investigated. Commercial MPM systems are currently being developed as a quality monitoring tool with the aim of detecting faulty parts already in the build process and, thus, reducing costs in LPBF. A detailed analysis of coupon specimens allowed two processing windows to be established for a suitably dense material at layer thicknesses of 30 µm and 50 µm, which were subsequently evaluated with two complex thermomechanical-fatigue (TMF) panels. Variations due to the location on the build platform were taken into account for the parameter development. Importantly, integrally averaged MPM intensities showed no direct correlation with total porosities, while the robustness of the melting process, impacted strongly by balling, affected the scattering of the MPM response and can thus be assessed. However, the MPM results, similar to material properties such as porosity, cannot be directly transferred from coupon specimens to components due to the influence of the local part geometry and heat transport on the build platform. Different MPM intensity ranges are obtained on cuboids and TMF panels despite similar LPBF parameters. Nonetheless, besides identifying LPBF parameter windows with a stable process, MPM allowed the successful detection of individual defects on the surface and in the bulk of the large demonstrators and appears to be a suitable tool for quality monitoring during fabrication and non-destructive evaluation of the LPBF process.
Language	English
Publishing date	2020-10-16
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2487261-1
ISSN	1996-1944
ISSN	1996-1944
DOI	10.3390/ma13204626
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Article ; Online: La plage urbaine spontanée, une nouvelle forme d’appropriation oisive de l’espace public

Maroua En-nejjari / Ignacio Requena / Daniel Siret

Les Cahiers de la Recherche Architecturale, Urbaine et Paysagère (2020)

2020

Abstract: Spontaneous urban beaches are contemporary public spaces where city dwellers behave as if they were on the beach, although these places have not been apparently planned for this purpose. Such urban public spaces, often located along the waterside, ... ...

Abstract	Spontaneous urban beaches are contemporary public spaces where city dwellers behave as if they were on the beach, although these places have not been apparently planned for this purpose. Such urban public spaces, often located along the waterside, reflect the emergence of a new approach to body and idleness in the public realm on the one hand, and the trend of functional diversion and spontaneous appropriation of the city on the other hand. This paper sheds light on the identification of the spatio-temporal, socio-cultural and political conditions of the emergence of a spontaneous beach situation in the urban environment. To do so, we have conducted an in-situ field survey in the city of Nantes in the summer of 2018, followed by a survey work out of beach situations, conducted with city dwellers and public officials of the city in 2019.
Keywords	Urban Beach ; Beach Situation ; Public Space ; Idleness ; Urban Ambiance ; Architecture ; NA1-9428 ; Geography. Anthropology. Recreation ; G
Subject code	720
Language	French
Publishing date	2020-05-01T00:00:00Z
Publisher	Ministère de la culture
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Efecto del aceite de Sacha Inchi (Plukenetiavolubilis L.) en la carcinogénesis de colon inducida por 1,2–dimetilhidrazina en ratas Holtzman.

Centurión-Rodríguez, César Abel / Huamán-Saavedra, Juan Jorge / Requena-Fuentes, Víctor Raúl

Revista de gastroenterologia del Peru : organo oficial de la Sociedad de Gastroenterologia del Peru

2017 Volume 37, Issue 2, Page(s) 129–136

Abstract: Objectives: To evaluate the preventive effect of Sacha Inchi oil (SIO) on 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis (CC) in Holtzman rats.: Materials and methods: Experimental study with 28 Holtzman male albino rats randomly ... ...

Title translation	Effect of Sacha Inchi oil (Plukenetiavolubilis L.) on 1,2-dimethylhydrazine-induced colon carcinogenesis in Holtzman rats.
Abstract	Objectives: To evaluate the preventive effect of Sacha Inchi oil (SIO) on 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis (CC) in Holtzman rats. Materials and methods: Experimental study with 28 Holtzman male albino rats randomly distributed into 4 groups: a positive control group exposed to DMH (C1), a negative control group exposed to SIO at 150 uL/kg/day (C2), and two experimental groups exposed to DMH with SIO at 150 uL/kg/day (E1) and SIO at 300 uL/kg/day (E2). The DMH was applied for 8 weeks and the total induction time was 22 weeks. Pathological examination was performed by identifying cancerous tumor lesions in the guts. The preventive effect was evaluated based on proportions of lack of lesion in the groups exposed to DMH. Results: Cancerous tumor lesions were identified in: two specimens of group C1, one specimen of group E1 and two specimens of group E2. No intestinal lesions were identified in group C2. The proportions of lack of lesion were: in group C1 of 75%, in group E1 of 87.5% and group E2 of 75%. No significant differences were found (p>0.05). Conclusions: It was not found a significant protective effect of SIO on DMH-induced CC in Holtzman rats, compared to control group.
Language	Spanish
Publishing date	2017-04
Publishing country	Peru
Document type	English Abstract ; Journal Article
ZDB-ID	2058591-3
ISSN	1609-722X ; 1022-5129
ISSN (online)	1609-722X
ISSN	1022-5129
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Book ; Online ; E-Book: Cutaneous adnexal neoplasms

Requena, Luis / Sangüeza, Omar P.

2017

Author's details	Luis Requena, Omar Sangüeza
Language	English
Size	1 Online-Ressource (xxiii, 1052 Seiten), Illustrationen
Publisher	Springer
Publishing place	Cham
Publishing country	Switzerland
Document type	Book ; Online ; E-Book
Remark	Zugriff für angemeldete ZB MED-Nutzerinnen und -Nutzer
HBZ-ID	HT019616481
ISBN	978-3-319-45704-8 ; 9783319457031 ; 3-319-45704-7 ; 3319457039
DOI	10.1007/978-3-319-45704-8
Database	ZB MED Catalogue: Medicine, Health, Nutrition, Environment, Agriculture

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Article: Study of the angular distribution of photon-induced Au-target L X-rays

Requena, S / Williams, S

Radiation physics and chemistry. 2011 May, v. 80, no. 5

2011

Abstract: We report the results of experiments involving the Lα and Lβ X-ray lines produced by 0–15keV bremsstrahlung incident on Au targets of various thicknesses at forward-scattered angles ranging from 25° to 155°. The relative intensities are compared to the ... ...

Abstract	We report the results of experiments involving the Lα and Lβ X-ray lines produced by 0–15keV bremsstrahlung incident on Au targets of various thicknesses at forward-scattered angles ranging from 25° to 155°. The relative intensities are compared to the predictions of the Monte Carlo code, PENELOPE. While the overall shapes of the angular distributions are similar to the distributions predicted by PENELOPE, several notable differences between our experimental results and the data generated by the program are observed.
Keywords	X-radiation ; gold ; prediction
Language	English
Dates of publication	2011-05
Size	p. 629-631.
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	2019621-0
ISSN	1878-1020 ; 0969-806X ; 1359-0197 ; 0146-5724
ISSN (online)	1878-1020
ISSN	0969-806X ; 1359-0197 ; 0146-5724
DOI	10.1016/j.radphyschem.2011.01.009
Database	NAL-Catalogue (AGRICOLA)

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Book: Cutaneous soft tissue tumors

Requena, Luis / Kutzner, Heinz

2015

Author's details	Luis Requena ; Heinz Kutzner
Keywords	Skin Neoplasms ; Neoplasms, Connective and Soft Tissue
Language	English
Size	XV, 963 S. : zahlr. Ill., graph. Darst.
Publisher	Wolters Kluwer
Publishing place	Philadelphia u.a.
Publishing country	United States
Document type	Book
Note	Includes bibliographical references
Accompanying material	Zugang zur Internetausgabe über Code
HBZ-ID	HT018403527
ISBN	978-1-4511-9276-6 ; 1-4511-9276-2
Database	Catalogue ZB MED Medicine, Health

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Shelf mark	Loan status	Location
2014 A 3684	available for loan (reading room)	Lesesaal EG
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Article ; Online: De novo assembly and functional annotation of Myrciaria dubia fruit transcriptome reveals multiple metabolic pathways for L-ascorbic acid biosynthesis.

Castro, Juan C / Maddox, J Dylan / Cobos, Marianela / Requena, David / Zimic, Mirko / Bombarely, Aureliano / Imán, Sixto A / Cerdeira, Luis A / Medina, Andersson E

BMC genomics

2015 Volume 16, Page(s) 997

Abstract: ... phytochemicals, but is best known by its high L-ascorbic acid (AsA) content in fruits. Pronounced variation ... identified: animal-like pathway, myo-inositol pathway, L-gulose pathway, D-mannose/L-galactose pathway, and ...

Abstract	Background: Myrciaria dubia is an Amazonian fruit shrub that produces numerous bioactive phytochemicals, but is best known by its high L-ascorbic acid (AsA) content in fruits. Pronounced variation in AsA content has been observed both within and among individuals, but the genetic factors responsible for this variation are largely unknown. The goals of this research, therefore, were to assemble, characterize, and annotate the fruit transcriptome of M. dubia in order to reconstruct metabolic pathways and determine if multiple pathways contribute to AsA biosynthesis. Results: In total 24,551,882 high-quality sequence reads were de novo assembled into 70,048 unigenes (mean length = 1150 bp, N50 = 1775 bp). Assembled sequences were annotated using BLASTX against public databases such as TAIR, GR-protein, FB, MGI, RGD, ZFIN, SGN, WB, TIGR_CMR, and JCVI-CMR with 75.2 % of unigenes having annotations. Of the three core GO annotation categories, biological processes comprised 53.6 % of the total assigned annotations, whereas cellular components and molecular functions comprised 23.3 and 23.1 %, respectively. Based on the KEGG pathway assignment of the functionally annotated transcripts, five metabolic pathways for AsA biosynthesis were identified: animal-like pathway, myo-inositol pathway, L-gulose pathway, D-mannose/L-galactose pathway, and uronic acid pathway. All transcripts coding enzymes involved in the ascorbate-glutathione cycle were also identified. Finally, we used the assembly to identified 6314 genic microsatellites and 23,481 high quality SNPs. Conclusions: This study describes the first next-generation sequencing effort and transcriptome annotation of a non-model Amazonian plant that is relevant for AsA production and other bioactive phytochemicals. Genes encoding key enzymes were successfully identified and metabolic pathways involved in biosynthesis of AsA, anthocyanins, and other metabolic pathways have been reconstructed. The identification of these genes and pathways is in agreement with the empirically observed capability of M. dubia to synthesize and accumulate AsA and other important molecules, and adds to our current knowledge of the molecular biology and biochemistry of their production in plants. By providing insights into the mechanisms underpinning these metabolic processes, these results can be used to direct efforts to genetically manipulate this organism in order to enhance the production of these bioactive phytochemicals. The accumulation of AsA precursor and discovery of genes associated with their biosynthesis and metabolism in M. dubia is intriguing and worthy of further investigation. The sequences and pathways produced here present the genetic framework required for further studies. Quantitative transcriptomics in concert with studies of the genome, proteome, and metabolome under conditions that stimulate production and accumulation of AsA and their precursors are needed to provide a more comprehensive view of how these pathways for AsA metabolism are regulated and linked in this species.
MeSH term(s)	Ascorbic Acid/biosynthesis ; Ascorbic Acid/genetics ; Biosynthetic Pathways ; High-Throughput Nucleotide Sequencing/methods ; M Phase Cell Cycle Checkpoints ; Molecular Sequence Annotation ; Myrtaceae/enzymology ; Myrtaceae/genetics ; Plant Proteins/genetics ; Sequence Analysis, RNA/methods ; Transcriptome
Chemical Substances	Plant Proteins ; Ascorbic Acid (PQ6CK8PD0R)
Language	English
Publishing date	2015-11-24
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	1471-2164
ISSN (online)	1471-2164
DOI	10.1186/s12864-015-2225-6
Database	MEDical Literature Analysis and Retrieval System OnLINE

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