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Article ; Online: Sporadic Amyotrophic Lateral Sclerosis Skeletal Muscle Transcriptome Analysis: A Comprehensive Examination of Differentially Expressed Genes.

Gascón, Elisa / Zaragoza, Pilar / Calvo, Ana Cristina / Osta, Rosario

2024 Volume 14, Issue 3

Abstract: Amyotrophic lateral sclerosis (ALS) that comprises sporadic (sALS) and familial (fALS) cases, is a devastating neurodegenerative disorder characterized by progressive degeneration of motor neurons, leading to muscle atrophy and various clinical ... ...

Abstract	Amyotrophic lateral sclerosis (ALS) that comprises sporadic (sALS) and familial (fALS) cases, is a devastating neurodegenerative disorder characterized by progressive degeneration of motor neurons, leading to muscle atrophy and various clinical manifestations. However, the complex underlying mechanisms affecting this disease are not yet known. On the other hand, there is also no good prognosis of the disease due to the lack of biomarkers and therapeutic targets. Therefore, in this study, by means of bioinformatics analysis, sALS-affected muscle tissue was analyzed using the GEO GSE41414 dataset, identifying 397 differentially expressed genes (DEGs). Functional analysis revealed 320 up-regulated DEGs associated with muscle development and 77 down-regulated DEGs linked to energy metabolism. Protein-protein interaction network analysis identified 20 hub genes, including
MeSH term(s)	Humans ; Transcriptome/genetics ; Amyotrophic Lateral Sclerosis/genetics ; Amyotrophic Lateral Sclerosis/metabolism ; MicroRNAs/genetics ; MicroRNAs/metabolism ; Muscle, Skeletal/metabolism ; Biomarkers
Chemical Substances	MicroRNAs ; Biomarkers ; MIRN206 microRNA, human
Language	English
Publishing date	2024-03-20
Publishing country	Switzerland
Document type	Review ; Journal Article
ZDB-ID	2701262-1
ISSN	2218-273X ; 2218-273X
ISSN (online)	2218-273X
ISSN	2218-273X
DOI	10.3390/biom14030377
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Susceptibility of Ovine Bone Marrow-Derived Mesenchymal Stem Cell Spheroids to Scrapie Prion Infection.

Hernaiz, Adelaida / Cobeta, Paula / Marín, Belén / Vázquez, Francisco José / Badiola, Juan José / Zaragoza, Pilar / Bolea, Rosa / Martín-Burriel, Inmaculada

Animals : an open access journal from MDPI

2023 Volume 13, Issue 6

Abstract: In neurodegenerative diseases, including prion diseases, cellular in vitro models appear as fundamental tools for the study of pathogenic mechanisms and potential therapeutic compounds. Two-dimensional (2D) monolayer cell culture systems are the most ... ...

Abstract	In neurodegenerative diseases, including prion diseases, cellular in vitro models appear as fundamental tools for the study of pathogenic mechanisms and potential therapeutic compounds. Two-dimensional (2D) monolayer cell culture systems are the most used cell-based assays, but these platforms are not able to reproduce the microenvironment of in vivo cells. This limitation can be surpassed using three-dimensional (3D) culture systems such as spheroids that more effectively mimic in vivo cell interactions. Herein, we evaluated the effect of scrapie prion infection in monolayer-cultured ovine bone marrow-derived mesenchymal stem cells (oBM-MSCs) and oBM-MSC-derived spheroids in growth and neurogenic conditions, analyzing their cell viability and their ability to maintain prion infection. An MTT assay was performed in oBM-MSCs and spheroids subjected to three conditions: inoculated with brain homogenate from scrapie-infected sheep, inoculated with brain homogenate from healthy sheep, and non-inoculated controls. The 3D conditions improved the cell viability in most cases, although in scrapie-infected spheroids in growth conditions, a decrease in cell viability was observed. The levels of pathological prion protein (PrP
Language	English
Publishing date	2023-03-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606558-7
ISSN	2076-2615
ISSN	2076-2615
DOI	10.3390/ani13061043
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Article ; Online: Novel polymorphisms in the prion protein gene (PRNP) and stability of the resultant prion protein in different horse breeds.

Sola, Diego / Artigas, Rody / Mediano, Diego R / Zaragoza, Pilar / Badiola, Juan José / Martín-Burriel, Inmaculada / Acín, Cristina

Veterinary research

2023 Volume 54, Issue 1, Page(s) 94

Abstract: Prion diseases are fatal neurodegenerative disorders in which the main pathogenic event is the conversion of the cellular prion protein ( ... ...

Abstract	Prion diseases are fatal neurodegenerative disorders in which the main pathogenic event is the conversion of the cellular prion protein (PrP
MeSH term(s)	Animals ; Amino Acid Sequence ; Horse Diseases/genetics ; Horses/genetics ; Polymorphism, Genetic ; Prion Diseases/genetics ; Prion Diseases/veterinary ; Prion Proteins/genetics ; Prion Proteins/metabolism ; Prions/genetics
Chemical Substances	Prion Proteins ; Prions
Language	English
Publishing date	2023-10-17
Publishing country	England
Document type	Journal Article
ZDB-ID	1146298-x
ISSN	1297-9716 ; 0928-4249
ISSN (online)	1297-9716
ISSN	0928-4249
DOI	10.1186/s13567-023-01211-8
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Article: Analysis of Plasma-Derived Exosomal MicroRNAs as Potential Biomarkers for Canine Idiopathic Epilepsy.

García-Gracia, Mireya / Moreno-Martinez, Laura / Hernaiz, Adelaida / Usón, Sebastián / Moral, Jon / Sanz-Rubio, David / Zaragoza, Pilar / Palacio, Jorge / Rosado, Belén / Osta, Rosario / García-Belenguer, Sylvia / Martín Burriel, Inmaculada

Animals : an open access journal from MDPI

2024 Volume 14, Issue 2

Abstract: Epilepsy is one of the most prevalent complex neurological diseases in both the canine and human species, with the idiopathic form as its most common diagnosis. MicroRNAs (miRNAs) are small, noncoding RNA molecules that play a role in gene regulation ... ...

Abstract	Epilepsy is one of the most prevalent complex neurological diseases in both the canine and human species, with the idiopathic form as its most common diagnosis. MicroRNAs (miRNAs) are small, noncoding RNA molecules that play a role in gene regulation processes and appear to be a promising biological target for convulsion control. These molecules have been reported as constituents of the internal content of exosomes, which are small extracellular vesicles released by cells. In this study, exosome samples were isolated from the plasma of 23 dogs, including 9 dogs with epilepsy responsive to treatment, 6 dogs with drug-resistant epilepsy, and 8 control dogs. Plasma exosomes were then characterized by electron transmission microscopy, nanoparticle tracking analysis, and dot blotting. Afterwards, the microRNA-enriched RNA content of exosomes was isolated, and miRNA quantification was performed by quantitative real-time PCR. Seven circulating miRNAs that have been previously described in the literature as potential diagnostic or prognostic biomarkers for epilepsy were evaluated. We observed significant differences in miR-16 (
Language	English
Publishing date	2024-01-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606558-7
ISSN	2076-2615
ISSN	2076-2615
DOI	10.3390/ani14020252
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Article ; Online: Susceptibility of Ovine Bone Marrow-Derived Mesenchymal Stem Cell Spheroids to Scrapie Prion Infection

Hernaiz, Adelaida / Cobeta, Paula / Marín, Belén / Vázquez, Francisco José / Badiola, Juan José / Zaragoza, Pilar / Bolea, Rosa / Martín-Burriel, Inmaculada

Animals. 2023 Mar. 13, v. 13, no. 6

2023

Abstract	In neurodegenerative diseases, including prion diseases, cellular in vitro models appear as fundamental tools for the study of pathogenic mechanisms and potential therapeutic compounds. Two-dimensional (2D) monolayer cell culture systems are the most used cell-based assays, but these platforms are not able to reproduce the microenvironment of in vivo cells. This limitation can be surpassed using three-dimensional (3D) culture systems such as spheroids that more effectively mimic in vivo cell interactions. Herein, we evaluated the effect of scrapie prion infection in monolayer-cultured ovine bone marrow-derived mesenchymal stem cells (oBM-MSCs) and oBM-MSC-derived spheroids in growth and neurogenic conditions, analyzing their cell viability and their ability to maintain prion infection. An MTT assay was performed in oBM-MSCs and spheroids subjected to three conditions: inoculated with brain homogenate from scrapie-infected sheep, inoculated with brain homogenate from healthy sheep, and non-inoculated controls. The 3D conditions improved the cell viability in most cases, although in scrapie-infected spheroids in growth conditions, a decrease in cell viability was observed. The levels of pathological prion protein (PrPSᶜ) in scrapie-infected oBM-MSCs and spheroids were measured by ELISA. In neurogenic conditions, monolayer cells and spheroids maintained the levels of PrPSᶜ over time. In growth conditions, however, oBM-MSCs showed decreasing levels of PrPSᶜ throughout time, whereas spheroids were able to maintain stable PrPSᶜ levels. The presence of PrPSᶜ in spheroids was also confirmed by immunocytochemistry. Altogether, these results show that a 3D culture microenvironment improves the permissiveness of oBM-MSCs to scrapie infection in growth conditions and maintains the infection ability in neurogenic conditions, making this model of potential use for prion studies.
Keywords	brain ; cell culture ; cell viability ; immunocytochemistry ; mesenchymal stromal cells ; models ; prions ; scrapie ; sheep ; therapeutics ; toxicity testing
Language	English
Dates of publication	2023-0313
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article ; Online
ZDB-ID	2606558-7
ISSN	2076-2615
ISSN	2076-2615
DOI	10.3390/ani13061043
Database	NAL-Catalogue (AGRICOLA)

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Article: Equine Mesenchymal Stem Cells Influence the Proliferative Response of Lymphocytes: Effect of Inflammation, Differentiation and MHC-Compatibility.

Cequier, Alina / Romero, Antonio / Vázquez, Francisco J / Vitoria, Arantza / Bernad, Elvira / Fuente, Sara / Zaragoza, Pilar / Rodellar, Clementina / Barrachina, Laura

Animals : an open access journal from MDPI

2022 Volume 12, Issue 8

Abstract: Immunomodulation and immunogenicity are pivotal aspects for the therapeutic use of mesenchymal stem cells (MSCs). Since the horse is highly valuable as both a patient and translational model, further knowledge on equine MSC immune properties is required. ...

Abstract	Immunomodulation and immunogenicity are pivotal aspects for the therapeutic use of mesenchymal stem cells (MSCs). Since the horse is highly valuable as both a patient and translational model, further knowledge on equine MSC immune properties is required. This study analysed how inflammation, chondrogenic differentiation and compatibility for the major histocompatibility complex (MHC) influence the MSC immunomodulatory-immunogenicity balance. Equine MSCs in basal conditions, pro-inflammatory primed (MSC-primed) or chondrogenically differentiated (MSC-chondro) were co-cultured with either autologous or allogeneic MHC-matched/mismatched lymphocytes in immune-suppressive assays (immunomodulation) and in modified one-way mixed leukocyte reactions (immunogenicity). After co-culture, frequency and proliferation of T cell subsets and B cells were assessed by flow cytometry and interferon-ɣ (IFNɣ) secretion by ELISA. MSC-primed showed higher regulatory potential by decreasing proliferation of cytotoxic and helper T cells and B cells. However, MHC-mismatched MSC-primed can also activate lymphocytes (proliferative response and IFNɣ secretion), likely due to increased MHC-expression. MSC-chondro maintained their regulatory ability and did not increase their immunogenicity, but showed less capacity than MSC-primed to induce regulatory T cells and further stimulated B cells. Subsequent in vivo studies are needed to elucidate the complex interactions between MSCs and the recipient immune system, which is critical to develop safe and effective therapies.
Language	English
Publishing date	2022-04-11
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606558-7
ISSN	2076-2615
ISSN	2076-2615
DOI	10.3390/ani12080984
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Article ; Online: 5-Methylcytosine and 5-Hydroxymethylcytosine in Scrapie-Infected Sheep and Mouse Brain Tissues.

Hernaiz, Adelaida / Sentre, Sara / Betancor, Marina / López-Pérez, Óscar / Salinas-Pena, Mónica / Zaragoza, Pilar / Badiola, Juan José / Toivonen, Janne Markus / Bolea, Rosa / Martín-Burriel, Inmaculada

International journal of molecular sciences

2023 Volume 24, Issue 2

Abstract: Scrapie is a neurodegenerative disorder belonging to the group of transmissible spongiform encephalopathies or prion diseases, which are caused by an infectious isoform of the innocuous cellular prion protein ( ... ...

Abstract	Scrapie is a neurodegenerative disorder belonging to the group of transmissible spongiform encephalopathies or prion diseases, which are caused by an infectious isoform of the innocuous cellular prion protein (PrP
MeSH term(s)	Animals ; Mice ; 5-Methylcytosine/metabolism ; Brain/metabolism ; Prion Diseases/genetics ; Prion Diseases/metabolism ; Prions/genetics ; Prions/metabolism ; Scrapie/genetics ; Scrapie/metabolism ; Sheep ; DNA Methylation/genetics ; DNA Methylation/physiology ; Epigenesis, Genetic/genetics ; Epigenesis, Genetic/physiology ; Histone Deacetylase 2/genetics ; Histone Deacetylase 2/metabolism ; DNA Methyltransferase 3B
Chemical Substances	5-hydroxymethylcytosine (1123-95-1) ; 5-Methylcytosine (6R795CQT4H) ; Prions ; TET1 protein, mouse ; Hdac1 protein, mouse (EC 3.5.1.98) ; Histone Deacetylase 2 (EC 3.5.1.98)
Language	English
Publishing date	2023-01-13
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms24021621
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Article: Genome-Wide Methylation Profiling in the Thalamus of Scrapie Sheep.

Hernaiz, Adelaida / Sanz, Arianne / Sentre, Sara / Ranera, Beatriz / Lopez-Pérez, Oscar / Zaragoza, Pilar / Badiola, Juan J / Filali, Hicham / Bolea, Rosa / Toivonen, Janne M / Martín-Burriel, Inmaculada

Frontiers in veterinary science

2022 Volume 9, Page(s) 824677

Abstract: Scrapie is a neurodegenerative disorder belonging to the group of transmissible spongiform encephalopathy (TSE). Scrapie occurs in sheep and goats, which are considered good natural animal models of these TSE. Changes in DNA methylation occur in the ... ...

Abstract	Scrapie is a neurodegenerative disorder belonging to the group of transmissible spongiform encephalopathy (TSE). Scrapie occurs in sheep and goats, which are considered good natural animal models of these TSE. Changes in DNA methylation occur in the central nervous system (CNS) of patients suffering from prion-like neurodegenerative diseases, such as Alzheimer's disease. Nevertheless, potential DNA methylation alterations have not yet been investigated in the CNS of any prion disease model or naturally infected cases, neither in humans nor in animals. Genome-wide DNA methylation patterns were studied in the thalamus obtained from sheep naturally infected with scrapie at a clinical stage (
Language	English
Publishing date	2022-02-16
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2834243-4
ISSN	2297-1769
ISSN	2297-1769
DOI	10.3389/fvets.2022.824677
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Article ; Online: Neuroprotective Fragment C of Tetanus Toxin Modulates IL-6 in an ALS Mouse Model.

Moreno-Martinez, Laura / de la Torre, Miriam / Muñoz, María J / Zaragoza, Pilar / Aguilera, José / Calvo, Ana C / Osta, Rosario

Toxins

2020 Volume 12, Issue 5

Abstract: Neuroinflammation plays a significant role in amyotrophic lateral sclerosis (ALS) pathology, leading to the development of therapies targeting inflammation in recent years. Our group has studied the tetanus toxin C-terminal fragment (TTC) as a ... ...

Abstract	Neuroinflammation plays a significant role in amyotrophic lateral sclerosis (ALS) pathology, leading to the development of therapies targeting inflammation in recent years. Our group has studied the tetanus toxin C-terminal fragment (TTC) as a therapeutic molecule, showing neuroprotective properties in the SOD1G93A mouse model. However, it is unknown whether TTC could have some effect on inflammation. The objective of this study was to assess the effect of TTC on the regulation of inflammatory mediators to elucidate its potential role in modulating inflammation occurring in ALS. After TTC treatment in SOD1G93A mice, levels of eotaxin-1, interleukin (IL)-2, IL-6 and macrophage inflammatory protein (MIP)-1 alpha (α) and galectin-1 were analyzed by immunoassays in plasma samples, whilst protein expression of caspase-1, IL-1β, IL-6 and NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) was measured in the spinal cord, extensor digitorum longus (EDL) muscle and soleus (SOL) muscle. The results showed reduced levels of IL-6 in spinal cord, EDL and SOL in treated SOD1G93A mice. In addition, TTC showed a different role in the modulation of NLRP3 and caspase-1 depending on the tissue analyzed. In conclusion, our results suggest that TTC could have a potential anti-inflammatory effect by reducing IL-6 levels in tissues drastically affected by the disease. However, further research is needed to study more in depth the anti-inflammatory effect of TTC in ALS.
MeSH term(s)	Amyotrophic Lateral Sclerosis/drug therapy ; Amyotrophic Lateral Sclerosis/genetics ; Amyotrophic Lateral Sclerosis/metabolism ; Animals ; Anti-Inflammatory Agents/pharmacology ; Caspase 1/metabolism ; Disease Models, Animal ; Down-Regulation ; Female ; Inflammasomes/metabolism ; Inflammation Mediators/metabolism ; Interleukin-6/metabolism ; Male ; Mice, Inbred C57BL ; Mice, Transgenic ; Muscle, Skeletal/drug effects ; Muscle, Skeletal/metabolism ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism ; Neuroprotective Agents/pharmacology ; Peptide Fragments/pharmacology ; Spinal Cord/drug effects ; Spinal Cord/metabolism ; Superoxide Dismutase-1/genetics ; Tetanus Toxin/pharmacology
Chemical Substances	Anti-Inflammatory Agents ; Inflammasomes ; Inflammation Mediators ; Interleukin-6 ; NLR Family, Pyrin Domain-Containing 3 Protein ; Neuroprotective Agents ; Nlrp3 protein, mouse ; Peptide Fragments ; Tetanus Toxin ; interleukin-6, mouse ; tetanus toxin fragment C ; Superoxide Dismutase-1 (EC 1.15.1.1) ; Casp1 protein, mouse (EC 3.4.22.36) ; Caspase 1 (EC 3.4.22.36)
Language	English
Publishing date	2020-05-17
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2518395-3
ISSN	2072-6651 ; 2072-6651
ISSN (online)	2072-6651
ISSN	2072-6651
DOI	10.3390/toxins12050330
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Allo-antibody production after intraarticular administration of mesenchymal stem cells (MSCs) in an equine osteoarthritis model: effect of repeated administration, MSC inflammatory stimulation, and equine leukocyte antigen (ELA) compatibility.

Barrachina, Laura / Cequier, Alina / Romero, Antonio / Vitoria, Arantza / Zaragoza, Pilar / Vázquez, Francisco José / Rodellar, Clementina

Stem cell research & therapy

2020 Volume 11, Issue 1, Page(s) 52

Abstract: Background: Antibody production after allogeneic administration of mesenchymal stem cells (MSCs) could impact their clinical application. Proinflammatory priming of MSCs can potentiate their regulatory ability in vivo but increased expression of major ... ...

Abstract	Background: Antibody production after allogeneic administration of mesenchymal stem cells (MSCs) could impact their clinical application. Proinflammatory priming of MSCs can potentiate their regulatory ability in vivo but increased expression of major histocompatibility complex (MHC) might augment their immunogenicity, potentially leading to immune memory thus limiting repeated allogeneic administration. This study aimed at evaluating the production of cytotoxic allo-antibodies directed against donor's ELA (equine leukocyte antigen) in mismatched and halfmatched horses receiving repeated intraarticular administration of stimulated MSCs (MSC-primed) and unstimulated MSCs (MSC-naïve) in pathologic joints. Methods: From available stored samples from a previous in vivo study, cells from one donor and serially collected sera (five time-points) from three groups of recipients were used based on their ELA haplotypes to perform microcytotoxicity assays: Group 1 recipients mismatched with the donor that received MSC-naïve (naïve-mismatched recipients); Group 2 recipients mismatched with the donor that received MSC-primed (primed-mismatched recipients); Group 3 recipients halfmatched with the donor (sharing 1/2 haplotypes) that received MSC-primed (primed-halfmatched recipients). Sera from recipients (neat, 1:2 and 1:16 dilution) were tested against target cells from the donor (cryopreserved and expanded MSC-naïve and MSC-primed) or from one animal presenting the same ELA haplotypes than the donor (fresh peripheral blood lymphocytes as control). Results: One to three weeks after first MSC administration, all recipient groups produced allo-antibodies regardless of MSC received (naïve or primed) and matching degree with donor. However, secondary response after MSC re-exposure was less evident in halfmatched recipients (MSC-primed) than in mismatched ones (both MSC-naïve and MSC-primed). Recipients of MSC-primed (both mismatched and halfmatched) tended towards developing lower antibody response than MSC-naïve recipients in vivo, but MSC-primed were targeted to death in higher percentage in vitro in the microcytoxicity assay. Conclusions: After first intraarticular allogeneic administration, the immunomodulatory profile of MSC-primed would have led to lower antibody production, but these antibodies would target more easily MSC-primed after second injection (re-exposure), likely because of their higher MHC expression.
MeSH term(s)	Animals ; Disease Models, Animal ; Female ; Horses ; Immunomodulation/immunology ; Leukocytes/immunology ; Male ; Mesenchymal Stem Cell Transplantation/methods ; Mesenchymal Stem Cells/metabolism ; Osteoarthritis
Language	English
Publishing date	2020-02-07
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2548671-8
ISSN	1757-6512 ; 1757-6512
ISSN (online)	1757-6512
ISSN	1757-6512
DOI	10.1186/s13287-020-1571-8
Database	MEDical Literature Analysis and Retrieval System OnLINE

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