Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda

Madrid, Spain

Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda

Madrid, Spain
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Perez-Luz S.,Autonomous University of Madrid | Perez-Luz S.,CIBER ISCIII | Perez-Luz S.,Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda | Gimenez-Cassina A.,Karolinska Institutet | And 7 more authors.
Genomics | Year: 2015

Friedreich's ataxia (FRDA) is the most common form of hereditary ataxia caused by recessive mutations in the FXN gene. Recent results have indicated the presence of different frataxin isoforms due to alternative gene expression mechanisms. Our previous studies demonstrated the advantages of using high-capacity herpes simplex virus type 1 (HSV-1) amplicon vectors containing the entire FXN genomic locus (iBAC-FXN) as a gene-delivery vehicle capable of ensuring physiologically-regulated and long-term persistence. Here we describe how expression from the 135. kb human FXN genomic locus produces the three frataxin isoforms both in cultured neuronal cells and also in vivo. Moreover, we also observed the correct expression of these frataxin isoforms in patient-derived cells after delivery of the iBAC-. FXN. These results lend further support to the potential use of HSV-1 vectors containing entire genomic loci whose expression is mediated by complex transcriptional and posttranscriptional mechanisms for gene therapy applications. © 2015 Elsevier Inc.


Citores M.J.,Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda Idiphim | Duca A.,Hospital Universitario Puerta Of Hierro Majadahonda | de la Fuente S.,Hospital Universitario Puerta Of Hierro Majadahonda | Vilches C.,Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda | And 2 more authors.
Clinical Transplantation | Year: 2016

Liver transplantation activates the innate immune system through toll-like receptors (TLRs), potentially leading to allograft rejection and graft failure. We evaluated the association of single-nucleotide polymorphisms in TLR genes with the severity of hepatitis C virus recurrence after liver transplantation (LT). This is a two-center study of 176 adult patients who received a first LT from deceased donors for hepatitis C virus (HCV) cirrhosis. Eleven polymorphisms were evaluated by real-time polymerase chain reaction and melting curves analyses: TLR1 (Asp248Ser and Ser602Ile), TLR2 (Arg753Gln), TLR3 (Leu412Phe), TLR4 (Asp299Gly), TLR5 (Arg392Stop), TLR6 (Ser249Pro), TLR7 (Gln11Leu), TLR8 (Met1Val), and TLR9 (−1237T/C and −1486C/T). The CC genotype of TLR3 Leu412Phe in liver recipients was associated with severe recurrence (odds ratio (OR) = 2.01, 95% confidence interval (95% CI) = 1.02-3.93, p = 0.04). We also analyzed this polymorphism in 72 of their donors but no association was found with severity of HCV recurrence (p = 0.89). Multivariate analysis showed donor age older than 40 yr (OR=2.93; 95% CI = 1.49–5.8, p = 0.002) and the TLR3 Leu412Phe CC genotype (OR=2.02, 95%CI=1.01–4.05, p = 0.046) were independently associated with severe HCV recurrence. Our results show that the TLR3 Leu412Phe CC genotype is independently associated with severity of hepatitis C recurrence after LT. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd


Gonzalo-Gobernado R.,Instituto Ramon Y Cajal Of Investigacion Sanitaria | Calatrava-Ferreras L.,Instituto Ramon Y Cajal Of Investigacion Sanitaria | Reimers D.,Instituto Ramon Y Cajal Of Investigacion Sanitaria | Herranz A.S.,Instituto Ramon Y Cajal Of Investigacion Sanitaria | And 5 more authors.
PLoS ONE | Year: 2013

Liver growth factor (LGF) is a hepatic mitogen purified some years ago that promotes proliferation of different cell types and the regeneration of damaged tissues, including brain tissue. Considering the possibility that LGF could be used as a therapeutic agent in Parkinson's disease, we analyzed its potential neuroregenerative and/or neuroprotective activity when peripherally administered to unilaterally 6-hydroxydopamine (6-OHDA)-lesioned rats. For these studies, rats subjected to nigrostriatal lesions were treated intraperitoneally twice a week with LGF (5 microg/rat) for 3 weeks. Animals were sacrificed 4 weeks after the last LGF treatment. The results show that LGF stimulates sprouting of tyrosine hydroxylase-positive terminals and increases tyrosine hydroxylase and dopamine transporter expression, as well as dopamine levels in the denervated striatum of 6-OHDA-lesioned rats. In this structure, LGF activates microglia and raises tumor necrosis factor-alpha protein levels, which have been reported to have a role in neuroregeneration and neuroprotection. Besides, LGF stimulates the phosphorylation of MAPK/ERK1/2 and CREB, and regulates the expression of proteins which are critical for cell survival such as Bcl2 and Akt. Because LGF partially protects dopamine neurons from 6-OHDA neurotoxicity in the substantia nigra, and reduces motor deficits in these animals, we propose LGF as a novel factor that may be useful in the treatment of Parkinson's disease. © 2013 Gonzalo-Gobernado et al.


Amigo-Jimenez I.,CSIC - Biological Research Center | Bailon E.,CSIC - Biological Research Center | Aguilera-Montilla N.,CSIC - Biological Research Center | Terol M.J.,Hospital Clinico Universitario | And 2 more authors.
Oncotarget | Year: 2015

CLL remains an incurable disease in spite of the many new compounds being studied. Arsenic trioxide (ATO) induces apoptosis in all CLL cell types and could constitute an efficient therapy. To further explore this, we have studied the influence of stromal cells, key components of the CLL microenvironment, on the response of CLL cells to ATO. Bone marrow stromal cells induced CLL cell resistance to 2 μM ATO and led to activation of Lyn, ERK, PI3K and PKC, as well as NF-κB and STAT3. Mcl-1, Bcl-xL, and Bfl-1 were also upregulated after the co-culture. Inhibition experiments indicated that PI3K and PKC were involved in the resistance to ATO induced by stroma. Moreover, idelalisib and sotrastaurin, specific inhibitors for PI3Kδ and PKCβ, respectively, inhibited Akt phosphorylation, NF-κB/STAT3 activation and Mcl-1 upregulation, and rendered cells sensitive to ATO. Mcl-1 was central to the mechanism of resistance to ATO, since: 1) Mcl-1 levels correlated with the CLL cell response to ATO, and 2) blocking Mcl-1 expression or function with specific siRNAs or inhibitors overcame the protecting effect of stroma. We have therefore identified the mechanism involved in the CLL cell resistance to ATO induced by bone marrow stroma and show that idelalisib or sotrastaurin block this mechanism and restore sensibility to ATO. Combination of ATO with these inhibitors may thus constitute an efficient treatment for CLL.


PubMed | CSIC - Biological Research Center, Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda and Hospital Clinico Universitario
Type: Journal Article | Journal: Oncotarget | Year: 2016

CLL remains an incurable disease in spite of the many new compounds being studied. Arsenic trioxide (ATO) induces apoptosis in all CLL cell types and could constitute an efficient therapy. To further explore this, we have studied the influence of stromal cells, key components of the CLL microenvironment, on the response of CLL cells to ATO. Bone marrow stromal cells induced CLL cell resistance to 2 M ATO and led to activation of Lyn, ERK, PI3K and PKC, as well as NF-B and STAT3. Mcl-1, Bcl-xL, and Bfl-1 were also upregulated after the co-culture. Inhibition experiments indicated that PI3K and PKC were involved in the resistance to ATO induced by stroma. Moreover, idelalisib and sotrastaurin, specific inhibitors for PI3K and PKC, respectively, inhibited Akt phosphorylation, NF-B/STAT3 activation and Mcl-1 upregulation, and rendered cells sensitive to ATO. Mcl-1 was central to the mechanism of resistance to ATO, since: 1) Mcl-1 levels correlated with the CLL cell response to ATO, and 2) blocking Mcl-1 expression or function with specific siRNAs or inhibitors overcame the protecting effect of stroma. We have therefore identified the mechanism involved in the CLL cell resistance to ATO induced by bone marrow stroma and show that idelalisib or sotrastaurin block this mechanism and restore sensibility to ATO. Combination of ATO with these inhibitors may thus constitute an efficient treatment for CLL.


PubMed | Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda
Type: Journal Article | Journal: Veterinary anaesthesia and analgesia | Year: 2016

To evaluate and compare three intravenous (IV) doses of a ketamine-propofol admixture (ketofol) for induction of anaesthesia in unpremedicated rabbits.Prospective, randomized blinded trial.Twenty-one healthy female New Zealand rabbits weighing 2.70.1kg.Animals were allocated randomly into three groups of seven animals and received 1 (KP1), 3 (KP3) or 5 (KP5) mgkg(-1) of both ketamine and propofol in a 1:1mgkg(-1) ratio admixture. Cardiorespiratory parameters and arterial blood gases were measured at baseline, 2 and 5minutes after drug administration. The time to loss of the righting reflex (LORR) and the duration of action and apnoea were recorded. The quality of induction and intubation were scored. Data were compared using a two-way anova or a t-test for unpaired data, as relevant.The time to LORR was the shortest (115seconds) and the duration of action the longest (37426seconds) in group KP5. Group KP1 did not lose the righting reflex; instead mild to moderate sedation was observed in this group. The quality of induction in group KP5 was smooth, but ranged from smooth to fair in group KP3. Intubation was not possible in the KP1 group, and 10 animals in the other two groups showed some resistance to intubation. At 2 and 5minutes, the pulse rate was significantly higher in all three groups compared with baseline, but no statistical differences were seen in arterial blood pressures. Hypoxaemia and dose-dependent respiratory depression were observed in all groups, with periods of apnoea in the KP5 group.The IV ketamine-propofol admixture had a dose-dependent effect. Haemodynamic function was well maintained in all groups but hypoxemia was observed at the highest doses and oxygen administration is recommended. Addition of premedication or topical lidocaine is advisable to make intubation easier.


PubMed | Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda
Type: Journal Article | Journal: Veterinary anaesthesia and analgesia | Year: 2016

To evaluate and compare the use of intramuscular (IM) premedication with dexmedetomidine in combination with ketamine or alfaxalone in pigs.Prospective, randomized, blinded trial.Fourteen healthy 2-month-old Landrace Large White pigs weighing 21.5 0.6 kg.Animals were distributed randomly into two groups: group KD (n = 7) was given 10 mg kg(-1) IM ketamine + 10 g kg(-1) IM dexmedetomidine; and group AD (n = 7) was given 5 mg kg(-1) IM alfaxalone + 10 g kg(-1) IM dexmedetomidine mixed in the same syringe. Pain on injection, degree of sedation and quality of induction were scored. The time from induction of anaesthesia to recumbency was recorded. Once pigs were recumbent, reflexes were evaluated. Pulse and respiratory rates, end-tidal carbon dioxide and arterial oxygen saturation were recorded at 5 and 10 minutes after drug administration. Data were compared using a two-way anova or a t-test for unpaired data as relevant. Data are presented as the mean standard deviation (range).Two animals in both groups showed slight pain on drug injection. The time to lateral recumbency in group KD [187 34 seconds (153-230)] was similar to group AD [206 36 seconds (150-248)]. In group AD, sedation was deeper, and the quality of anaesthetic induction was smoother. When moved for anaesthesia, five pigs in group KD vocalized. There were no differences between groups in pulse rates, arterial oxygen saturation and end-tidal carbon dioxide; however, the respiratory rate at 10 minutes was significantly higher in group KD than in group AD.IM dexmedetomidine in combination with ketamine in pigs induced moderate to deep sedation and fair to smooth induction of anaesthesia. When dexmedetomidine was combined with alfaxalone, sedation was deeper, and induction was of a better quality.


PubMed | CSIC - Biological Research Center and Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda
Type: Journal Article | Journal: Oncotarget | Year: 2016

CLL remains an incurable disease in spite of the many new compounds being tested. Arsenic trioxide (ATO) induces apoptosis in all CLL cell types and could constitute an efficient therapy. To further explore this, we have studied the gene expression profile induced by ATO in CLL cells. ATO modulated many genes, largely involved in oxidative stress, being HMOX1 the most upregulated gene, also induced at the protein level. ATO also increased MMP-9, as we previously observed, both at the mRNA and protein level. Using specific inhibitors, qPCR analyses, and gene silencing approaches we demonstrate that upregulation of MMP-9 by ATO involved activation of the p38 MAPK/AP-1 signaling pathway. Moreover, gene silencing HMOX1 or inhibiting HMOX1 activity enhanced p38 MAPK phosphorylation and c-jun expression/activation, resulting in transcriptional upregulation of MMP-9. Overexpression of HMOX1 or enhancement of its activity, had the opposite effect. Cell viability analyses upon modulation of HMOX1 expression or activity demonstrated that HMOX1 had a pro-apoptotic role and enhanced the cytotoxic effect of ATO in CLL cells. We have therefore identified a new mechanism in which HMOX1 plays a central role in the response of CLL cells to ATO and in the regulation of the anti-apoptotic protein MMP-9. Thus, HMOX1 arises as a new therapeutic target in CLL and the combination of HMOX1 modulators with ATO may constitute an efficient therapeutic strategy in CLL.


PubMed | Hospital Universitario Puerta Of Hierro Majadahonda, Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda and Institute Investigacion Sanitaria Puerta Of Hierro Majadahonda Idiphim
Type: Journal Article | Journal: Clinical transplantation | Year: 2016

Liver transplantation activates the innate immune system through toll-like receptors (TLRs), potentially leading to allograft rejection and graft failure. We evaluated the association of single-nucleotide polymorphisms in TLR genes with the severity of hepatitis C virus recurrence after liver transplantation (LT). This is a two-center study of 176 adult patients who received a first LT from deceased donors for hepatitis C virus (HCV) cirrhosis. Eleven polymorphisms were evaluated by real-time polymerase chain reaction and melting curves analyses: TLR1 (Asp248Ser and Ser602Ile), TLR2 (Arg753Gln), TLR3 (Leu412Phe), TLR4 (Asp299Gly), TLR5 (Arg392Stop), TLR6 (Ser249Pro), TLR7 (Gln11Leu), TLR8 (Met1Val), and TLR9 (-1237T/C and -1486C/T). The CC genotype of TLR3 Leu412Phe in liver recipients was associated with severe recurrence (odds ratio (OR) = 2.01, 95% confidence interval (95% CI) = 1.02-3.93, p = 0.04). We also analyzed this polymorphism in 72 of their donors but no association was found with severity of HCV recurrence (p = 0.89). Multivariate analysis showed donor age older than 40 yr (OR=2.93; 95% CI = 1.49-5.8, p = 0.002) and the TLR3 Leu412Phe CC genotype (OR=2.02, 95%CI=1.01-4.05, p = 0.046) were independently associated with severe HCV recurrence. Our results show that the TLR3 Leu412Phe CC genotype is independently associated with severity of hepatitis C recurrence after LT.

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