Publications by authors named "Marta Soler"

76 Publications

Changes in Renal Resistive Index Values in Healthy Puppies during the First Months of Life.

Animals (Basel) 2020 Aug 3;10(8). Epub 2020 Aug 3.

Department of Animal Medicine and Surgery, Veterinary Faculty, University of Murcia, 30100 Murcia, Spain.

The purpose of this study is to establish renal resistive index (RRI) of normal kidneys in puppies aged from newborn to 20 weeks of age and to determine the age at which RRI reaches adult dog values. Six healthy adult intact beagles and six puppies from 1 day after birth to 20 weeks of age were used. In the adult dogs, the ultrasonographic scans were performed once, and in the puppies, the ultrasonographic studies were performed on the first day after birth and at 1, 2, 3, 4, 6, 8, 12, 16, and 20 weeks of age. RRI was obtained at the interlobular and arcuate arteries in each kidney. There were no statistical differences between the RRI values obtained between the right and left kidney nor between intrarenal arteries (interlobar and arcuate). The RRI was the highest during the first weeks of life, after which it declined gradually with increasing age reaching adult dog values at 12 weeks of age. In conclusion, the normal mean RRI is age dependent in dogs. Twelve weeks can be regarded as the age at which adult mean RRI criteria can be applied to puppies.
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http://dx.doi.org/10.3390/ani10081338DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7459847PMC
August 2020

Short-Term Effects of Deliberate Subparaneural or Subepineural Injections With Saline Solution or Bupivacaine 0.75% in the Sciatic Nerve of Rabbits.

Front Vet Sci 2020 12;7:217. Epub 2020 May 12.

Department of Medicine and Animal Surgery, Faculty of Veterinary Science, University of Murcia, Murcia, Spain.

Ultrasound (US)-guided techniques for peripheral nerve blockade have revealed that intraneural injections are relatively frequent and not necessarily associated with neurological deficits. To evaluate the short-term effects of deliberate injections performed under direct vision in two different sites of the sciatic nerve (ScN). Seventy-two New Zealand white rabbits randomly assigned to one of four experimental groups ( = 18) were employed. All procedures were conducted at a proximal femoral level where the ScN incorporates the common peroneal nerve and the tibial nerve (TN). Fixed volumes of 0.5 ml of saline solution (ES group) or bupivacaine 0.75% (EB group) were administered extrafascicularly inside the paraneurium of the ScN or intrafascicularly (IS and IB groups) under the epineurium of the TN. Cross-sectional area (CSA) and relative echogenicity (RE) of the entire ScN were determined by US before injections, after injections, and at 3 and 7 days. ScN samples were obtained for structural and ultrastructural histopathological studies. Proprioceptive, sensorial, and motor function were clinically evaluated on a daily basis. The CSA of the ScN increased significantly immediately after injections when compared with pre-injection values in all groups ( < 0.05). The RE of the ScN decreased in relation to pre-injection values in all groups ( < 0.05). The CSA and RE of the ScN returned to normal values 7 days after injections in almost all groups. Injected nerves showed histological signs of mild perineural inflammation. Histopathological scores were not significantly different between groups ( > 0.05). The architecture of the ScN was preserved in all rabbits at 3 days and in 31/32 rabbits at 7 days. A focal area of damaged nerve fibers with degeneration of the axons and myelin sheath affecting the TN was observed in one rabbit of the IB group. Nerve function was not clinically impaired in any case. Despite the lack of severe nerve disruption observed in most rabbits, the evidence of a focal area of damaged nerve fibers in one rabbit injected intrafascicularly with bupivacaine confirms that intrafascicular injections should be avoided as they may increase the risk of nerve damage.
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http://dx.doi.org/10.3389/fvets.2020.00217DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7235316PMC
May 2020

Ultrasonographic measurement of adrenal gland-to-aorta ratio as a method of estimating adrenal size in dogs.

Vet Rec 2020 06 14;186(19):e27. Epub 2019 Nov 14.

Department of Medicine and Surgery, University of Murcia, Murcia, Spain.

Background: Adrenal size has been used as the principal criterion for differentiating a normal gland from adrenal hyperplasia. The objectives of this study were to establish an ultrasonographic measurement of adrenal gland-to-aorta (adrenal/Ao) ratio to estimate the adrenal size and to assess the effects of bodyweight, age and sex on the adrenal/Ao ratio in non-adrenal gland disease dogs.

Methods: Two hundred and thirty-four dogs (120 entire females and 114 entire males) considered non-adrenal gland disease were included in this study. Dogs were allocated into three bodyweight categories (<10 kg, 10-20 kg and >20 kg), and four age groups (<1 year, 1-5 years, 5-10 years and >10 years old). Measurements of the maximal thickness of caudal pole of both adrenal glands and the aortic luminal diameter in sagittal plane were performed on the ultrasonographic images. Three different ratios were calculated for each dog.

Results: Sex and age did not influence on the adrenal/Ao ratio. There were differences (P<0.05) between the three dog sizes for adrenal/Ao ratio, being the highest value for small size and the lowest value for large size.

Conclusions: In this study, only bodyweight influences the ultrasound measurement of the adrenal/Ao ratio in non-adrenal gland disease dogs.
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http://dx.doi.org/10.1136/vr.105188DOI Listing
June 2020

B-cell leukemia transdifferentiation to macrophage involves reconfiguration of DNA methylation for long-range regulation.

Leukemia 2020 04 12;34(4):1158-1162. Epub 2019 Nov 12.

Josep Carreras Leukaemia Research Institute (IJC), Badalona, Barcelona, Catalonia, Spain.

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http://dx.doi.org/10.1038/s41375-019-0643-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7214273PMC
April 2020

The transcribed pseudogene RPSAP52 enhances the oncofetal HMGA2-IGF2BP2-RAS axis through LIN28B-dependent and independent let-7 inhibition.

Nat Commun 2019 09 4;10(1):3979. Epub 2019 Sep 4.

Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), L'Hospitalet de Llobregat, Barcelona, Catalonia, Spain.

One largely unknown question in cell biology is the discrimination between inconsequential and functional transcriptional events with relevant regulatory functions. Here, we find that the oncofetal HMGA2 gene is aberrantly reexpressed in many tumor types together with its antisense transcribed pseudogene RPSAP52. RPSAP52 is abundantly present in the cytoplasm, where it interacts with the RNA binding protein IGF2BP2/IMP2, facilitating its binding to mRNA targets, promoting their translation by mediating their recruitment on polysomes and enhancing proliferative and self-renewal pathways. Notably, downregulation of RPSAP52 impairs the balance between the oncogene LIN28B and the tumor suppressor let-7 family of miRNAs, inhibits cellular proliferation and migration in vitro and slows down tumor growth in vivo. In addition, high levels of RPSAP52 in patient samples associate with a worse prognosis in sarcomas. Overall, we reveal the roles of a transcribed pseudogene that may display properties of an oncofetal master regulator in human cancers.
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http://dx.doi.org/10.1038/s41467-019-11910-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6726650PMC
September 2019

Epigenetic loss of RNA-methyltransferase NSUN5 in glioma targets ribosomes to drive a stress adaptive translational program.

Acta Neuropathol 2019 12 19;138(6):1053-1074. Epub 2019 Aug 19.

Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), L'Hospitalet, Barcelona, Catalonia, Spain.

Tumors have aberrant proteomes that often do not match their corresponding transcriptome profiles. One possible cause of this discrepancy is the existence of aberrant RNA modification landscapes in the so-called epitranscriptome. Here, we report that human glioma cells undergo DNA methylation-associated epigenetic silencing of NSUN5, a candidate RNA methyltransferase for 5-methylcytosine. In this setting, NSUN5 exhibits tumor-suppressor characteristics in vivo glioma models. We also found that NSUN5 loss generates an unmethylated status at the C3782 position of 28S rRNA that drives an overall depletion of protein synthesis, and leads to the emergence of an adaptive translational program for survival under conditions of cellular stress. Interestingly, NSUN5 epigenetic inactivation also renders these gliomas sensitive to bioactivatable substrates of the stress-related enzyme NQO1. Most importantly, NSUN5 epigenetic inactivation is a hallmark of glioma patients with long-term survival for this otherwise devastating disease.
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http://dx.doi.org/10.1007/s00401-019-02062-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6851045PMC
December 2019

Cholangiocarcinoma and hepatic myelolipoma incarcerated in a peritoneopericardial diaphragmatic hernia with pulmonary metastasis and carcinomatosis in a cat.

JFMS Open Rep 2019 Jan-Jun;5(1):2055116919835081. Epub 2019 Feb 26.

Veterinary Teaching Hospital, University of Murcia, Murcia, Spain.

Case Summary: A 14-year-old female neutered Persian-cross cat was presented with a 1 week history of anorexia and lethargy. On physical examination, marked tachypnoea and dyspnoea were evident. Radiographs of the thorax revealed a globoid-shaped cardiac silhouette with heterogeneous opacity consistent with a peritoneopericardial diaphragmatic hernia (PPDH), pulmonary nodules compatible with metastasis, seven sternal segments and a small liver in the cranial abdomen with loss of serosal detail. On echocardiography, there was no evidence of cardiac tamponade. Triple-phase CT angiography demonstrated a mixed soft tissue-, mineral- and fat-attenuated liver mass arising from the left hepatic lobes that showed a pronounced heterogeneous contrast-enhancement pattern within the pericardial sac, which was producing a marked mass effect on the adjacent structures. Additionally, there was an increase in attenuation of the mesenteric fat and peritoneal effusion. The pulmonary nodules were confirmed. Imaging findings were compatible with a malignant hepatic neoplasia incarcerated in a PPDH, lung metastasis and carcinomatosis. Owing to the poor prognosis, the cat was humanely euthanased. Histopathological diagnosis was cholangiocellular carcinoma and hepatic myelolipoma, pulmonary metastasis and carcinomatosis.

Relevance And Novel Information: Hepatic cholangiocarcinoma incarcerated in a PPDH with pulmonary metastasis and carcinomatosis has not been previously described. Suspicion of a hepatic neoplasia should be raised in cases of PPDH and pulmonary nodules.
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http://dx.doi.org/10.1177/2055116919835081DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6393819PMC
February 2019

Effects of midazolam before or after alfaxalone for co-induction of anaesthesia in healthy dogs.

Vet Anaesth Analg 2018 Sep 19;45(5):609-617. Epub 2018 Jun 19.

Department of Animal Medicine and Surgery, Faculty of Veterinary, University of Murcia, Murcia, Spain.

Objective: To study the effect of alternating the order of midazolam and alfaxalone administration on the incidence of behavioural changes, alfaxalone induction dose and some cardiorespiratory variables in healthy dogs.

Study Design: Prospective, randomized, controlled, clinical trial.

Animals: A total of 33 client-owned dogs undergoing elective procedures.

Methods: Following intramuscular acepromazine (0.02 mg kg) and morphine (0.4 mg kg) premedication, anaesthesia was induced intravenously (IV) with a co-induction of either midazolam (0.25 mg kg) prior to alfaxalone (0.5 mg kg; group MA), or alfaxalone followed by midazolam at identical doses (group AM). The control group (CA) was administered normal saline IV prior to alfaxalone administration. Additional alfaxalone (0.25 mg kg increments) was administered as required in all groups until orotracheal intubation was possible. Changes in behaviour, quality of induction, ease of intubation and incidence of adverse events at induction were recorded. Heart rate (HR), respiratory rate (f) and systolic arterial blood pressure (SAP) were measured before treatments (baseline values), 30 minutes after premedication and at 0, 2, 5 and 10 minutes postintubation.

Results: The incidence of excitement was higher in group MA compared with groups CA (p=0.005) and AM (p=0.013). The mean induction dose of alfaxalone was lower in group AM compared with group CA (p=0.003). Quality of induction and ease of intubation were similar among groups. Mean HR values decreased after premedication and increased after alfaxalone administration in all groups. Mean SAP values were similar between groups. The number of animals that required manual ventilation was higher in the MA group.

Conclusions And Clinical Relevance: Despite a lower occurrence of adverse events at induction in group AM compared with group MA and a reduction of alfaxalone dose requirement in group AM compared with group CA, the use of an alfaxalone-midazolam co-induction does not seem to produce any cardiovascular or respiratory benefits in healthy dogs.
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http://dx.doi.org/10.1016/j.vaa.2018.04.002DOI Listing
September 2018

Circular RNA CpG island hypermethylation-associated silencing in human cancer.

Oncotarget 2018 Jun 26;9(49):29208-29219. Epub 2018 Jun 26.

Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona, Catalonia, Spain.

Noncoding RNAs (ncRNAs), such as microRNAs and long noncoding RNAs (lncRNAs), participate in cellular transformation. Work done in the last decade has also demonstrated that ncRNAs with growth-inhibitory functions can undergo promoter CpG island hypermethylation-associated silencing in tumorigenesis. Herein, we wondered whether circular RNAs (circRNAs), a type of RNA transcripts lacking 5'-3' ends and forming closed loops that are gaining relevance in cancer biology, are also a target of epigenetic inactivation in tumors. To tackle this issue, we have used cancer cells genetically deficient for the DNA methyltransferase enzymes in conjuction with circRNA expression microarrays. We have found that the loss of DNA methylation provokes a release of circRNA silencing. In particular, we have identified that promoter CpG island hypermethylation of the genes TUSC3 (tumor suppressor candidate 3), POMT1 (protein O-mannosyltransferase 1), ATRNL1 (attractin-like 1) and SAMD4A (sterile alpha motif domain containing 4A) is linked to the transcriptional downregulation of both linear mRNA and the hosted circRNA. Although some circRNAs regulate the linear transcript, we did not observe changes in TUSC3 mRNA levels upon TUSC3 circ104557 overexpression. Interestingly, we found circRNA-mediated regulation of target miRNAs and an growth inhibitory effect upon TUSC3 circ104557 transduction. Data mining for 5'-end CpG island methylation of TUSC3, ATRNL1, POMT1 and SAMD4A in cancer cell lines and primary tumors showed that the epigenetic defect was commonly observed among different tumor types in association with the diminished expression of the corresponding transcript. Our findings support a role for circRNA DNA methylation-associated loss in human cancer.
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http://dx.doi.org/10.18632/oncotarget.25673DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6044373PMC
June 2018

Inhibition of Gsk3b Reduces Nfkb1 Signaling and Rescues Synaptic Activity to Improve the Rett Syndrome Phenotype in Mecp2-Knockout Mice.

Cell Rep 2018 05;23(6):1665-1677

Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), 08908 L'Hospitalet, Barcelona, Catalonia, Spain; Institucio Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Catalonia, Spain; Physiological Sciences Department, School of Medicine and Health Sciences, University of Barcelona (UB), 08907 Catalonia, Spain. Electronic address:

Rett syndrome (RTT) is the second leading cause of mental impairment in girls and is currently untreatable. RTT is caused, in more than 95% of cases, by loss-of-function mutations in the methyl CpG-binding protein 2 gene (MeCP2). We propose here a molecular target involved in RTT: the glycogen synthase kinase-3b (Gsk3b) pathway. Gsk3b activity is deregulated in Mecp2-knockout (KO) mice models, and SB216763, a specific inhibitor, is able to alleviate the clinical symptoms with consequences at the molecular and cellular levels. In vivo, inhibition of Gsk3b prolongs the lifespan of Mecp2-KO mice and reduces motor deficits. At the molecular level, SB216763 rescues dendritic networks and spine density, while inducing changes in the properties of excitatory synapses. Gsk3b inhibition can also decrease the nuclear activity of the Nfkb1 pathway and neuroinflammation. Altogether, our findings indicate that Mecp2 deficiency in the RTT mouse model is partially rescued following treatment with SB216763.
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http://dx.doi.org/10.1016/j.celrep.2018.04.010DOI Listing
May 2018

Bromodomain inhibition shows antitumoral activity in mice and human luminal breast cancer.

Oncotarget 2017 Aug 29;8(31):51621-51629. Epub 2017 May 29.

Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona, Catalonia, Spain.

BET bromodomain inhibitors, which have an antitumoral effect against various solid cancer tumor types, have not been studied in detail in luminal breast cancer, despite the prevalence of this subtype of mammary malignancy. Here we demonstrate that the BET bromodomain inhibitor JQ1 exerts growth-inhibitory activity in human luminal breast cancer cell lines associated with a depletion of the C-MYC oncogene, but does not alter the expression levels of the BRD4 bromodomain protein. Interestingly, expression microarray analyses indicate that, upon JQ1 administration, the antitumoral phenotype also involves downregulation of relevant breast cancer oncogenes such as the Breast Carcinoma-Amplified Sequence 1 (BCAS1) and the PDZ Domain-Containing 1 (PDZK1). We have also applied these findings in an model by studying a transgenic mouse model representing the luminal B subtype of breast cancer, the MMTV-PyMT, in which the mouse mammary tumor virus promoter is used to drive the expression of the polyoma virus middle T-antigen to the mammary gland. We have observed that the use of the BET bromodomain inhibitor for the treatment of established breast neoplasms developed in the MMTV-PyMT model shows antitumor potential. Most importantly, if JQ1 is given before the expected time of tumor detection in the MMTV-PyMT mice, it retards the onset of the disease and increases the survival of these animals. Thus, our findings indicate that the use of bromodomain inhibitors is of great potential in the treatment of luminal breast cancer and merits further investigation.
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http://dx.doi.org/10.18632/oncotarget.18255DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5584274PMC
August 2017

Radiographic, ultrasonographic, and computed tomographic characteristics of an accessory liver lobe in a cat.

Vet Radiol Ultrasound 2019 May 6;60(3):E29-E32. Epub 2017 Aug 6.

Department of Animal Medicine and Surgery, University of Murcia, Espinardo, Murcia, 30100, Spain.

A 5-year-old male Norwegian Forest cat presented with increased hepatic serum biochemical parameters. Abdominal radiography showed an oval cranioventral mass and ultrasound revealed a mobile mass attached to one hepatic lobe. Computed tomography (CT) confirmed that the mass was attached to the right medial liver lobe. Differential diagnoses were an accessory liver lobe, benign neoplasia, and focal nodular hyperplasia. The mass was removed and histopathology confirmed the mass to be normal liver tissue. Accessory liver lobe should be included in the differential diagnosis of a mobile cranial abdominal mass with a similar ultrasonographic or CT appearance to the liver.
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http://dx.doi.org/10.1111/vru.12538DOI Listing
May 2019

Increased inflammatory effect of electronegative LDL and decreased protection by HDL in type 2 diabetic patients.

Atherosclerosis 2017 10 13;265:292-298. Epub 2017 Jul 13.

Cardiovascular Biochemistry, Biomedical Research Institute Sant Pau (IIB-Sant Pau), C/Sant Antoni M. Claret 167, 08025 Barcelona, Spain; Molecular Biology and Biochemistry Department, Universitat Autònoma de Barcelona (UAB) Faculty of Medicine, Building M. Cerdanyola del Vallès, Spain. Electronic address:

Background And Aims: Type 2 diabetic patients have an increased proportion of electronegative low-density lipoprotein (LDL(-)), an inflammatory LDL subfraction present in blood, and dysfunctional high-density lipoprotein (HDL). We aimed at examining the inflammatory effect of LDL(-) on monocytes and the counteracting effect of HDL in the context of type 2 diabetes.

Methods: This was a cross-sectional study in which the population comprised 3 groups (n = 12 in each group): type 2 diabetic patients with good glycaemic control (GC-T2DM patients), type 2 diabetic patients with poor glycaemic control (PC-T2DM), and a control group. Total LDL, HDL, and monocytes were isolated from plasma of these subjects. LDL(-) was isolated from total LDL by anion-exchange chromatography. LDL(-) from the three groups of subjects was added to monocytes in the presence or absence of HDL, and cytokines released by monocytes were quantified by ELISA.

Results: LDL(-) proportion and plasma inflammatory markers were increased in PC-T2DM patients. LDL(-) from PC-T2DM patients induced the highest IL1β, IL6, and IL10 release in monocytes compared to LDL(-) from GC-T2DM and healthy subjects, and presented the highest content of non-esterified fatty acids (NEFA). In turn, HDL from PC-T2DM patients showed the lowest ability to inhibit LDL(-)-induced cytokine release in parallel to an impaired ability to decrease NEFA content in LDL(-).

Conclusions: Our findings show an imbalance in the pro- and anti-inflammatory effects of lipoproteins from T2DM patients, particularly in PC-T2DM.
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http://dx.doi.org/10.1016/j.atherosclerosis.2017.07.015DOI Listing
October 2017

A Cu/Cu prototypical organometallic mechanism for the deactivation of an active pincer-like Cu catalyst in Ullmann-type couplings.

Chem Commun (Camb) 2017 Aug 21;53(62):8786-8789. Epub 2017 Jul 21.

Institut de Química Computacional i Catàlisi (IQCC) and Departament de Química, Universitat de Girona, Campus de Montilivi, E-17003 Girona, Catalonia, Spain.

Unraveling the mechanistic details of copper-catalyzed arylation of nucleophiles (Ullmann-type couplings) is a very challenging task. It is a matter of intense debate whether it is a radical-based process or an organometallic redox-based process. The ancillary ligand choice in Ullmann-type couplings plays a key role in such transformations and can strongly influence the catalytic efficiency as well as the mechanism. Here, we show how a predesigned tridentate pincer-like catalyst undergoes a deactivation pathway through a Cu/Cu prototypical mechanism as demonstrated by helium-tagging infrared photodissociation (IRPD) spectroscopy and DFT studies, lending a strong support to the existence of an aryl-Cu species in the Ullmann couplings using this tridentate ligand.
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http://dx.doi.org/10.1039/c7cc04491gDOI Listing
August 2017

Detection and Characterization of R Loop Structures.

Methods Mol Biol 2017 ;1543:231-242

Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona, Catalonia, Spain.

R loops are special three stranded nucleic acid structures that comprise a nascent RNA hybridized with the DNA template strand, leaving a non-template DNA single-stranded. More specifically, R loops form in vivo as G-rich RNA transcripts invade the DNA duplex and anneal to the template strand to generate an RNA:DNA hybrid, leaving the non-template, G-rich DNA strand in a largely single-stranded conformation (Aguilera and Garcia-Muse, Mol Cell 46:115-124, 2012).DNA-RNA hybrids are a natural occurrence within eukaryotic cells, with levels of these hybrids increasing at sites with high transcriptional activity, such as during transcription initiation, repression, and elongation. RNA-DNA hybrids influence genomic instability, and growing evidence points to an important role for R loops in active gene expression regulation (Ginno et al., Mol Cell 45, 814-825, 2012; Sun et al., Science 340: 619-621, 2013; Bhatia et al., Nature 511, 362-365, 2014). Analysis of the occurrence of such structures is therefore of increasing relevance and herein we describe methods for the in vivo and in vitro identification and characterization of R loops in mammalian systems.R loops (DNA:RNA hybrids and the associated single-stranded DNA) have been traditionally associated with threats to genome integrity, making some regions of the genome more prone to DNA-damaging and mutagenic agents. Initially considered to be rare byproducts of transcription, over the last decade accumulating evidence has pointed to a new view in which R loops form more frequently than previously thought. The R loop field has become an increasingly expanded area of research, placing these structures as a major threat to genome stability but also as potential regulators of gene expression. Special interest has arisen as they have also been linked to a variety of diseases, including neurological disorders and cancer, positioning them as potential therapeutic targets [5].
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http://dx.doi.org/10.1007/978-1-4939-6716-2_13DOI Listing
February 2018

RNA-FISH to Study Regulatory RNA at the Site of Transcription.

Methods Mol Biol 2017 ;1543:221-229

Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona, Catalonia, Spain.

The increasing role of all types of regulatory RNAs in the orchestration of cellular programs has enhanced the development of a variety of techniques that allow its precise detection, quantification, and functional scrutiny. Recent advances in imaging and fluoresecent in situ hybridization (FISH) methods have enabled the utilization of user-friendly protocols that provide highly sensitive and accurate detection of ribonucleic acid molecules at both the single cell and subcellular levels. We herein describe the approach originally developed by Stellaris, in which the target RNA molecule is fluoresecently labeled with multiple tiled complementary probes each carrying a fluorophore, thus improving sensitivity and reducing the chance of false positives. We have applied this method to the detection of nascent RNAs that partake of special regulatory structures called R loops. Their growing role in active gene expression regulation (Aguilera and Garcia-Muse, Mol Cell 46:115-124, 2012; Ginno et al., Mol Cell 45:814-825, 2012; Sun et al., Science 340:619-621, 2013; Bhatia et al., Nature 511:362-365, 2014) imposes the use of a combination of in vivo and in vitro techniques for the detailed analysis of the transcripts involved. Therefore, their study is a good example to illustrate how RNA FISH, combined with transcriptional arrest and/or cell synchronization, permits localization and temporal characterization of potentially regulatory RNA sequences.
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http://dx.doi.org/10.1007/978-1-4939-6716-2_12DOI Listing
February 2018

IMAGING DIAGNOSIS-RADIOGRAPHIC, ULTRASONOGRAPHIC, AND COMPUTED TOMOGRAPHIC CHARACTERISTICS OF A DUODENAL DUPLICATION CYST IN A YOUNG CAT.

Vet Radiol Ultrasound 2018 May 29;59(3):E22-E27. Epub 2017 Jan 29.

Department of Animal Veterinary Medicine and Surgery, University of Murcia, Spain.

A 7-month-old, 2.8 kg, intact female Siamese cat was evaluated for repetitive and intermittent episodes of vomiting and anorexia. Abdominal palpation revealed a round, firm, nonpainful mass in the right cranial abdomen. Ultrasonography findings were consistent with a cystic structure adjacent to the descending duodenum. The structure exhibited a "muscular rim sign." A duodenal duplication cyst was confirmed by histopathological analysis. Computed tomography ruled out concurrent vertebral anomalies and clarified anatomic relationships for surgical planning. To the authors' knowledge, this is the first description of an ultrasound "muscular rim sign" in a duodenal duplication cyst in a cat.
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http://dx.doi.org/10.1111/vru.12469DOI Listing
May 2018

Epigenetic inactivation of the p53-induced long noncoding RNA TP53 target 1 in human cancer.

Proc Natl Acad Sci U S A 2016 11 7;113(47):E7535-E7544. Epub 2016 Nov 7.

Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), L'Hospitalet de Llobregat, 08908 Barcelona, Catalonia, Spain;

Long noncoding RNAs (lncRNAs) are important regulators of cellular homeostasis. However, their contribution to the cancer phenotype still needs to be established. Herein, we have identified a p53-induced lncRNA, TP53TG1, that undergoes cancer-specific promoter hypermethylation-associated silencing. In vitro and in vivo assays identify a tumor-suppressor activity for TP53TG1 and a role in the p53 response to DNA damage. Importantly, we show that TP53TG1 binds to the multifaceted DNA/RNA binding protein YBX1 to prevent its nuclear localization and thus the YBX1-mediated activation of oncogenes. TP53TG1 epigenetic inactivation in cancer cells releases the transcriptional repression of YBX1-targeted growth-promoting genes and creates a chemoresistant tumor. TP53TG1 hypermethylation in primary tumors is shown to be associated with poor outcome. The epigenetic loss of TP53TG1 therefore represents an altered event in an lncRNA that is linked to classical tumoral pathways, such as p53 signaling, but is also connected to regulatory networks of the cancer cell.
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http://dx.doi.org/10.1073/pnas.1608585113DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5127373PMC
November 2016

Comparison between ultrasonographic findings of benign and malignant canine mammary gland tumours using B-mode, colour Doppler, power Doppler and spectral Doppler.

Res Vet Sci 2016 Aug 31;107:141-146. Epub 2016 May 31.

Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Murcia University, 30100 Espinardo, Murcia, Spain.

The aim of this study was to evaluate whether the comparison between the ultrasonographic features of canine mammary tumours, assessed by B-Mode, colour Doppler, power Doppler, spectral Doppler, and histopathologic features, would help to differentiate if a tumour is benign or malignant. Ultrasonographic examinations of 104 tumours were performed. Volume, margins, presence of a capsule, echotexture and presence and distribution of the vascular flow of the tumours were evaluated. All the tumours were surgically removed, submitted for histopathologic examination and classified in two groups: Group I (benign tumours) and Group II (malignant tumours). Echotexture was the only parameter evaluated by B-Mode ultrasonography where significant differences were found (p<0.01), with tumours in Group I being homogeneous and tumours in Group II presenting greater heterogeneity. Presence of vascular flow was observed in most of the tumours from both groups and no differences between them were found. Regarding flow distribution, significant differences were observed between groups (p<0.05). In benign tumours, the most common vascular pattern was the peripheral, showing significant differences (p<0.05) compared to mixed and central patterns. In malignant tumours the mixed pattern was the most frequent. Also significant differences among other patterns (peripheral and central) were found. Concerning vascular resistivity and pulsatility indexes, there were no significant differences between the two groups. The echotexture and type of vascular flow pattern of canine mammary gland tumours may help, in a first examination of the tumour, to differentiate between benign and malignant tumours; however to reach a definitive diagnosis histological study is required.
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http://dx.doi.org/10.1016/j.rvsc.2016.05.015DOI Listing
August 2016

Orthogonal Discrimination among Functional Groups in Ullmann-Type C-O and C-N Couplings.

J Org Chem 2016 09 9;81(17):7315-25. Epub 2016 Jun 9.

Institut de Química Computacional i Catàlisi (IQCC) and Departament de Química, Universitat de Girona, Campus de Montilivi , E-17003 Girona, Catalonia, Spain.

The copper-catalyzed arylation of nucleophiles has been established as an efficient methodology for the formation of C-C and C-heteroatom bonds. Considering the advances during the last two decades, the ligand choice plays a key role in such transformations and can strongly influence the catalytic efficiency. The applicability of these Ullmann-type coupling reactions regarding the orthogonal selectivity of different functional groups constitutes a challenging subject for current synthetic strategies. Herein, we report a useful toolkit of Cu-based catalysts for the chemoselective arylation of a wide-range of nucleophiles in competitive reactions using aryl iodides and bromides. We show in this work that the arylation of all kinds of amides can be orthogonal to that of amines (aliphatic or aromatic) and phenol derivatives. This high chemoselectivity can be governed by the use of different ligands, yielding the desired coupling products under mild conditions. The selectivity trends are maintained for electronically biased iodobenzene and bromobenzene electrophiles. Radical clock experiments discard the occurrence of radical-based mechanisms.
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http://dx.doi.org/10.1021/acs.joc.6b01035DOI Listing
September 2016

Peptide-mediated vectorization of metal complexes: conjugation strategies and biomedical applications.

Dalton Trans 2016 Aug;45(33):12970-82

QBIS-CAT Research Group, Institut de Química Computacional i Catàlisi (IQCC) and Departament de Química, Universitat de Girona, Campus Montilivi, E-17071 Girona, Catalonia, Spain.

The rich chemical and structural versatility of transition metal complexes provides numerous novel paths to be pursued in the design of molecules that exert particular chemical or physicochemical effects that could operate over specific biological targets. However, the poor cell permeability of metallodrugs represents an important barrier for their therapeutic use. The conjugation between metal complexes and a functional peptide vector can be regarded as a versatile and potential strategy to improve their bioavailability and accumulation inside cells, and the site selectivity of their effect. This perspective lies in reviewing the recent advances in the design of metallopeptide conjugates for biomedical applications. Additionally, we highlight the studies where this approach has been directed towards the incorporation of redox active metal centers into living organisms for modulating the cellular redox balance, as a tool with application in anticancer therapy.
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http://dx.doi.org/10.1039/c5dt04529kDOI Listing
August 2016

Evaluation of an ultrasound-guided technique for axillary brachial plexus blockade in cats.

J Feline Med Surg 2017 02 10;19(2):146-152. Epub 2016 Jul 10.

1 Department of Animal Medicine and Surgery, University of Murcia, Murcia, Spain.

Objectives The aim of this study was to evaluate and refine an ultrasound (US)-guided technique to block the brachial plexus (BP) at the level of the axillary space in live cats. Methods Eight adult experimental cats were enrolled into the study. The animals were sedated and positioned in dorsal recumbency with the limb to be blocked abducted 90º. The US transducer was placed in the axillary region and a non-traumatic peripheral nerve block needle was inserted in-plane with respect to the transducer, medial to the BP up to the level of the axillary artery. Lidocaine 1% (0.4 ml/kg) was injected as the needle was being progressively withdrawn in a caudal-to-cranial direction. The efficacy of the block was confirmed by evaluation of the motor and sensory functions of the blocked forelimb. Motor blockade was assessed observing the position of the blocked leg on standing and walking patterns. Sensory blockade was evaluated by the stimulation of mechanical nociceptors in the dermatomes supplied by the four major sensory nerves of the distal thoracic limb. Results The BP was successfully located by US in all cases. The achieved BP block was complete in six cats (75%) and partial in the remaining two cats (25%). All animals recovered uneventfully from the sedation and the BP blocks. Conclusions and relevance The US-guided block at the axillary space evaluated in this study is a feasible, reproducible and safe technique to block the BP plexus in experimental live cats.
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http://dx.doi.org/10.1177/1098612X15618703DOI Listing
February 2017

IMAGING DIAGNOSIS-URETHROVAGINAL FISTULA CAUSED BY A MIGRATING GRASS AWN IN THE VAGINA.

Vet Radiol Ultrasound 2016 May 23;57(3):E30-3. Epub 2015 Nov 23.

Department of Animal Veterinary Medicine and Surgery, Veterinary Teaching Hospital, University of Murcia, 30100, Espinardo, Murcia, Spain.

A young intact female dog was presented with urinary incontinence. Abdominal ultrasound revealed the presence of hyperechoic linear structures within the cranial vagina suggestive of foreign material. A computed tomography (CT) retrograde vaginourethrogram demonstrated the presence of a fistulous tract between the urethra and vagina. A presumptive diagnosis of urethrovaginal fistula due to migration of foreign material was made. The grass awn was removed with vaginoscopic-guided retrieval. Fourteen days later, surgical repair of the fistula and an ovariohysterectomy were done. This case report emphasizes the usefulness of CT for diagnosis and precise anatomical localization of genitourinary tract fistulas.
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http://dx.doi.org/10.1111/vru.12311DOI Listing
May 2016

Design, Preparation, and Characterization of Zn and Cu Metallopeptides Based On Tetradentate Aminopyridine Ligands Showing Enhanced DNA Cleavage Activity.

Inorg Chem 2015 Nov 27;54(22):10542-58. Epub 2015 Oct 27.

QBIS-CAT Research Group, Institut de Química Computacional i Catàlisi (IQCC) and Departament de Química, Universitat de Girona , Campus Montilivi, E-17071 Girona, Catalonia, Spain.

The conjugation of redox-active complexes that can function as chemical nucleases to cationic tetrapeptides is pursued in this work in order to explore the expected synergistic effect between these two elements in DNA oxidative cleavage. Coordination complexes of biologically relevant first row metal ions, such as Zn(II) or Cu(II), containing the tetradentate ligands 1,4-dimethyl-7-(2-pyridylmethyl)-1,4,7-triazacyclononane ((Me2)PyTACN) and (2S,2S')-1,1'-bis(pyrid-2-ylmethyl)-2,2'-bipyrrolidine ((S,S')-BPBP) have been linked to a cationic LKKL tetrapeptide sequence. Solid-phase synthesis of the peptide-tetradentate ligand conjugates has been developed, and the preparation and characterization of the corresponding metallotetrapeptides is described. The DNA cleavage activity of Cu and Zn metallopeptides has been evaluated and compared to their metal binding conjugates as well as to the parent complexes and ligands. Very interestingly, the oxidative Cu metallopeptides 1Cu and 2Cu show an enhanced activity compared to the parent complexes, [Cu(PyTACN)](2+) and [Cu(BPBP)](2+), respectively. Under optimized conditions, 1Cu displays an apparent pseudo first-order rate constant (kobs) of ∼0.16 min(-1) with a supercoiled DNA half-life time (t1/2) of ∼4.3 min. On the other hand, kobs for 2Cu has been found to be ∼0.11 min(-1) with t1/2 ≈ 6.4 min. Hence, these results point out that the DNA cleavage activities promoted by the metallopeptides 1Cu and 2Cu render ∼4-fold and ∼23 rate accelerations in comparison with their parent Cu complexes. Additional binding assays and mechanistic studies demonstrate that the enhanced cleavage activities are explained by the presence of the cationic LKKL tetrapeptide sequence, which induces an improved binding affinity to the DNA, thus bringing the metal ion, which is responsible for cleavage, in close proximity.
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http://dx.doi.org/10.1021/acs.inorgchem.5b01680DOI Listing
November 2015

KAT6B Is a Tumor Suppressor Histone H3 Lysine 23 Acetyltransferase Undergoing Genomic Loss in Small Cell Lung Cancer.

Cancer Res 2015 Sep 24;75(18):3936-45. Epub 2015 Jul 24.

Cancer Epigenetics Group, Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), Barcelona, Catalonia, Spain. Department of Physiological Sciences II, School of Medicine, University of Barcelona, Barcelona, Catalonia, Spain. Institucio Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Catalonia, Spain.

Recent efforts to sequence human cancer genomes have highlighted that point mutations in genes involved in the epigenetic setting occur in tumor cells. Small cell lung cancer (SCLC) is an aggressive tumor with poor prognosis, where little is known about the genetic events related to its development. Herein, we have identified the presence of homozygous deletions of the candidate histone acetyltransferase KAT6B, and the loss of the corresponding transcript, in SCLC cell lines and primary tumors. Furthermore, we show, in vitro and in vivo, that the depletion of KAT6B expression enhances cancer growth, while its restoration induces tumor suppressor-like features. Most importantly, we demonstrate that KAT6B exerts its tumor-inhibitory role through a newly defined type of histone H3 Lys23 acetyltransferase activity.
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http://dx.doi.org/10.1158/0008-5472.CAN-14-3702DOI Listing
September 2015

Validation of the dorsal approach for the blockade of the femoral nerve using ultrasound and nerve electrolocation in cats.

J Feline Med Surg 2016 08 5;18(8):620-5. Epub 2015 Jun 5.

Department of Animal Medicine and Surgery, University of Murcia, Murcia, Spain.

Objectives: This study was conducted to validate the dorsal approach for femoral nerve (FN) blockade in cats and to verify the efficacy of the sole use of peripheral nerve electrolocation (PNE) or ultrasound (US)-guided technique to achieve the block.

Methods: This study was carried out in two phases. In phase 1, five adult experimental cats were used to validate the approach. In each cat, one FN was located by US and the accuracy of this location confirmed by PNE. Then, 2 mg/kg lidocaine 2% (diluted in saline to a final volume of 1 ml) was injected around the target nerve and the success of the blockade was evaluated. In phase 2, four adult experimental cats were included in two groups to verify the reliability of this approach to block eight FNs by the sole use of PNE (group 1) or US-guided technique (group 2). Evidence of motor blockade, time required to perform the blockade, onset time and duration of the blockades were determined.

Results: The FN was successfully located by US in all cats enrolled in phase 1, as confirmed by PNE in all cases. The success rate was clinically higher in group 2 (87.5%) than in group 1 (75.0%). The US-guided technique required less time to perform and produced blocks of longer duration. Recovery was uneventful in all cases.

Conclusions And Relevance: The combined use of PNE and US-guided technique enabled validation of the dorsal approach for the FN blockade as it provided a successful FN blockade in all cases. The sole use of a US-guided technique may offer some advantages over the use of a sole PNE-guided technique to perform these blocks.
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http://dx.doi.org/10.1177/1098612X15590868DOI Listing
August 2016

Head-to-head antisense transcription and R-loop formation promotes transcriptional activation.

Proc Natl Acad Sci U S A 2015 May 22;112(18):5785-90. Epub 2015 Apr 22.

Cancer Epigenetics and Biology Program, and

The mechanisms used by antisense transcripts to regulate their corresponding sense mRNAs are not fully understood. Herein, we have addressed this issue for the vimentin (VIM) gene, a member of the intermediate filament family involved in cell and tissue integrity that is deregulated in different types of cancer. VIM mRNA levels are positively correlated with the expression of a previously uncharacterized head-to-head antisense transcript, both transcripts being silenced in colon primary tumors concomitant with promoter hypermethylation. Furthermore, antisense transcription promotes formation of an R-loop structure that can be disfavored in vitro and in vivo by ribonuclease H1 overexpression, resulting in VIM down-regulation. Antisense knockdown and R-loop destabilization both result in chromatin compaction around the VIM promoter and a reduction in the binding of transcriptional activators of the NF-κB pathway. These results are the first examples to our knowledge of R-loop-mediated enhancement of gene expression involving head-to-head antisense transcription at a cancer-related locus.
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http://dx.doi.org/10.1073/pnas.1421197112DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4426458PMC
May 2015

Enzyme-triggered delivery of chlorambucil from conjugates based on the cell-penetrating peptide BP16.

Org Biomol Chem 2015 Feb;13(5):1470-80

QBIS-CAT Research Group, Institut de Química Computacional i Catàlisi (IQCC) and Departament de Química, Universitat de Girona, Campus Montilivi, E-17071 Girona, Catalonia, Spain.

The undecapeptide KKLFKKILKKL-NH2 (BP16) is a non-toxic cell-penetrating peptide (CPP) that is mainly internalized into cancer cells through a clathrin dependent endocytic mechanism and localizes in late endosomes. Moreover, this CPP is able to enhance the cellular uptake of chlorambucil (CLB) improving its cytotoxicity. In this work, we further explored the cell-penetrating properties of BP16 and those of its arginine analogue BP308. We investigated the influence on the cytotoxicity and on the cellular uptake of conjugating CLB at the N- or the C-terminal end of these undecapeptides. The effect of incorporating the cathepsin B-cleavable sequence Gly-Phe-Leu-Gly in CLB-BP16 and CLB-BP308 conjugates was also evaluated. The activity of CLB was significantly improved when conjugated at the N- or the C-terminus of BP16, or at the N-terminus of BP308. While CLB alone was not active (IC50 of 73.7 to >100 μM), the resulting conjugates displayed cytotoxic activity against CAPAN-1, MCF-7, PC-3, 1BR3G and SKMEL-28 cell lines with IC50 values ranging from 8.7 to 25.5 μM. These results were consistent with the internalization properties observed for the corresponding 5(6)-carboxyfluorescein-labeled conjugates. The presence of the tetrapeptide Gly-Phe-Leu-Gly at either the N- or the C-terminus of CLB-BP16 conjugates further increased the efficacy of CLB (IC50 of 3.6 to 16.2 μM), which could be attributed to its selective release in the lysosomal compartment. Enzymatic assays with cathepsin B showed the release of CLB-Gly-OH from these sequences within a short time. Therefore, the combination of BP16 with an enzymatic cleavable sequence can be used as a drug delivery system for the effective uptake and release of drugs in cancer cells.
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http://dx.doi.org/10.1039/c4ob01875cDOI Listing
February 2015

Prostacyclin-synthase expression in head and neck carcinoma patients and its prognostic value in the response to radiotherapy.

J Pathol 2015 Jan;235(1):125-35

Laboratory of Angiology, Vascular Biology and Inflammation, Institute of Biomedical Research (IIB Sant Pau) and Universitat Autònoma de Barcelona, Barcelona, Spain.

Prostacyclin (PGI2 ) plays a role in cancer progression but the mechanism is currently poorly understood. Additionally, no data are available about the prognostic value of the PGI2 pathway in head and neck squamous cell carcinoma (HNSCC) therapy. We evaluated the expression of the PGI2 pathway in HNSCC patients. PGI2 production and PGI synthase (PGIS) expression, in terms of mRNA (RT-PCR) and protein (immunoblotting), were lower in tumour samples than in non-tumoural mucosa, whereas, as expected, COX-2 expression was increased in HNSCC tumour samples. Using local control of the tumour after radiotherapy or chemoradiotherapy as a dependent variable, patients were classified into two categories of PGIS transcript levels. The high-PGIS group had a significantly lower frequency of local and distant failure than the low-PGIS group, and the 5-year cancer-specific survival was higher [90.2% (95% CI 81.0-99.4%) versus 60.5% (95% CI 44.4-76.6%)]. None of the four HNSCC cell lines analysed expressed PGIS and therefore they did not produce PGI2 . However, HNSCC-conditioned media enhanced PGI2 production in endothelial cells (ECs). The stable analogue of PGI2 , carbaprostacyclin (cPGI2 ), exerted little effect on HNSCC cell line migration, and no effect on cell cycle distribution or proliferation rate after radiation injury was observed. Nevertheless, cPGI2 promoted EP-4-dependent in vitro angiogenesis. Von Willebrand factor expression (EC marker) and capillary density were significantly higher in the group of patients with high expression of PGIS. Our results indicate that PGIS expression was associated with radiotherapy efficiency. Although we do not provide direct evidence of a relationship between tumour vascularization and radiotherapy efficiency, our results suggest that the effect of PGI2 is related to its ability to promote vascularization. These results also support the concept that co-adjuvant therapy with PGIS enhancers, such as retinoids, could have therapeutic value for HNSCC treatment.
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http://dx.doi.org/10.1002/path.4453DOI Listing
January 2015