Efecto de la hormona de crecimiento recombinante humana en la regeneración de nervio periférico: Trabajo experimental en el nervio cubital de la rata

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Cirugía. Fecha de lectura: 24-06-2016Cuando se producen lesiones en los nervios periféricos se pueden utilizar diferentes sustancias o productos para intentar mejorar su crecimiento y reparación. En el presente trabajo valoramos la efectividad de la hormona de crecimiento recombinante humana (Somatropina, GH) en la lesión del nervio periférico de la rata Wistar. Mediante técnica microquirúrgica se disecaba el nervio cubital en parte proximal de la extremidad superior de la rata. Posteriormente se seccionaba el nervio, suturando el cabo proximal y distal a un tubo de PVC para que regenerase a través de éste. Se han estudiado 32 nervios cubitales, obteniendo de forma aleatorizada: 18 nervios que han regenerado bajo el efecto de la hormona de crecimiento y 14 nervios sin la acción de la hormona. Se ha llevado a cabo un seguimiento durante 8 semanas, realizando electroneurografía de superficie cada 1-2 semanas tras la intervención. Hemos encontrado en el grupo que recibió la hormona: una mejoría significativa en la recuperación de la velocidad de conducción y un mayor incremento gradual en la amplitud del potencial motor a partir de la quinta semana, respecto al grupo que no se le administró la hormona. En el estudio histológico realizado al final del estudio, encontramos en el grupo de ratas a las que se les administró la hormona: una mejor arquitectura del nervio regenerado, una mayor densidad de fibras nerviosas, una mayor mielinización con menor grado de fibrosis y tejido de granulación endoneura

HGUE-C-1 is an atypical and novel colon carcinoma cell line

Background: Colorectal carcinoma is a common cause of cancer. Adjuvant treatments include: 5-fluorouracil administered together with folinic acid, or more recently, oral fluoropyrimidines such as capecitabine, in combination with oxaliplatin or irinotecan. Metastatic colorectal cancer patients can benefit from other additional treatments such as cetuximab or bevacizumab. Methods: Using cell culture techniques, we isolated clonal populations from primary cultures of ascitic effusion derived from a colon cancer patient and after several passages an established cell line, HGUE-C-1, was obtained. Genetic analysis of HGUE-C-1 cells was performed by PCR of selected exons and sequencing. Cell proliferation studies were performed by MTT assays and cell cycle analyses were performed by flow cytometry. Retinoblastoma activity was measured by luciferase assays and proteins levels and activity were analysed by Western blot or immunohistochemistry. Results: We have established a new cell line from ascitic efussion of a colon cancer patient who did not respond to 5-fluorouracil or irinotecan. HGUE-C-1 cells did not show microsatellite instability and did not harbour mutations in KRAS, BRAF, PI3KCA or TP53. However, these cells showed loss of heterozygosity affecting Adenomatous Polyposis Coli and nuclear staining of β-catenin protein. The HGUE-C-1 cell line was sensitive to erlotinib, gefitinib, NVP-BEZ235, rapamycin and trichostatin, among other drugs, but partially resistant to heat shock protein inhibitors and highly resistant to AZD-6244 and oxaliplatin, even though the patient from which this cell line was derived had not been exposed to these drugs. Molecular characterization of this cell line revealed low expression levels and activity of Retinoblastoma protein and elevated basal levels of Erk1/2 activity and p70S6K expression and activity, which may be related to chemoresistance mechanisms. Conclusions: HGUE-C-1 represents a novel and peculiar colon carcinoma model to study chemoresistance to chemotherapeutic agents and to novel anti-neoplasic drugs that interrupt signalling pathways such as the APC/βcatenin, Ras/Raf/Mek/Erk, PI3K/mTOR/p70S6K pathways as well as histone regulation mechanisms.This article has been funded by grants from the Instituto de Salud Carlos III FIS PI080901and FIS PI01202025 to Miguel Saceda

Differential Effects of IGF-1R Small Molecule Tyrosine Kinase Inhibitors BMS-754807 and OSI-906 on Human Cancer Cell Lines

We have determined the effects of the IGF-1R tyrosine kinase inhibitors BMS-754807 (BMS) and OSI-906 (OSI) on cell proliferation and cell-cycle phase distribution in human colon, pancreatic carcinoma, and glioblastoma cell lines and primary cultures. IGF-1R signaling was blocked by BMS and OSI at equivalent doses, although both inhibitors exhibited differential antiproliferative effects. In all pancreatic carcinoma cell lines tested, BMS exerted a strong antiproliferative effect, whereas OSI had a minimal effect. Similar results were obtained on glioblastoma primary cultures, where HGUE-GB-15, -16 and -17 displayed resistance to OSI effects, whereas they were inhibited in their proliferation by BMS. Differential effects of BMS and OSI were also observed in colon carcinoma cell lines. Both inhibitors also showed different effects on cell cycle phase distribution, BMS induced G2/M arrest followed by cell death, while OSI induced G1 arrest with no cell death. Both inhibitors also showed different effects on other protein kinases activities. Taken together, our results are indicative that BMS mainly acts through off-target effects exerted on other protein kinases. Given that BMS exhibits a potent antiproliferative effect, we believe that this compound could be useful for the treatment of different types of tumors independently of their IGF-1R activation status.This research was funded by a Grant from Instituto de Salud Carlos III Grant PI012/02025 co-supported by FEDER funds and PRECIPITA crowdfunding platform from Fundación Española para la Ciencia y la Tecnología (Fecyt) to M. Saceda and AMACMED (Asociación de mujeres afectadas por cáncer de mama de Elche y Comarca) and Monica Moraleda donation to M. Saceda. The Spanish Ministry of Economy and Competitiveness (MINECO, Project RTI2018-096724-B-C21) and the Generalitat Valenciana (PROMETEO/2016/006) supported the work in the Encinar laboratory

Crónicas de Jurisprudencia. Derechos Fundamentales y Libertades Púlbicas

Crónica de jurisprudencia sobre derechos y libertades de los siguientes tribunales: Tribunal Europeo de Derechos Humanos, Tribunal de Justicia de la Unión Europea, Tribunal Constitucional, Tribunal SupremoFUNDACIÓ BOSCH I GIMPERA y Editorial Aranzadi (Thomson Reuters)2021-2

Crónicas de Jurisprudencia. Derechos Fundamentales y Libertades Púlbicas

Crónica de la jurisprudencia sobre derechos y libertades de los siguientes tribunales: Tribunal Europeo de Derechos Humanos, Tribunal de Justicia de la Unión Europea, Tribunal Constitucional, Tribunal SupremoFUNDACIÓ BOSCH I GIMPERA y Editorial Aranzadi (Thomson Reuters)2020-2

Crónicas de Jurisprudencia. Derechos Fundamentales y Libertades Púlbicas

Cónica de la jurisprudencia sobre derechos y libertades de los siguientes tribunales: Tribunal Europeo de Derechos Humanos, Tribunal de Justicia de la Unión Europea, Tribunal Constitucional, Tribunal SupremoFUNDACIÓ BOSCH I GIMPERA y Editorial Aranzadi (Thomson Reuters)2021-2

Crónicas de Jurisprudencia. Derechos Fundamentales y Libertades Púlbicas

Crónica de jurisprudencia sobre derechos y libertades de los siguientes tribunales: Tribunal Europeo de Derechos Humanos, Tribunal de Justicia de la Unión Europea, Tribunal Constitucional, Tribunal SupremoFUNDACIÓ BOSCH I GIMPERA y Editorial Aranzadi (Thomson Reuters)2021-2

A case of acute spinal intradural hematoma due to spinal anesthesia

Spinal intradural hematoma is a rare complication of diagnostic lumbar puncture or spinal anesthesia. This complication could be overlooked with devastating neurological consequences due to a delay in diagnosis. Here, we reported a case of a patient with a lumbar spinal intradural hematoma as a result of a difficult spinal anesthesia

Dual regulation of P-glycoprotein expression by Trichostatin A in cancer cell lines

Background. It has been reported that the histone deacetylase inhibitor (iHDAc) trichostatin A (TSA) induces an increase in MDR1 gene transcription (ABCB1). This result would compromise the use of iHDACs in combination with other cytotoxic agents that are substrates of P-glycoprotein (Pgp). It has also been reported the use of alternative promoters by the ABCB1 gene and the existence of a traslational control of Pgp protein. Finally, the ABCB1 gene is located in a genetic locus with the nested gene RUNDC3B in the complementary DNA strand, raising the possibility that RUNDC3B expression could interfere with ABCB1 alternative promoter regulation. Methods. A combination of RT-PCR, real time RT-PCR, Western blot and drug accumulation assays by flow cytometry have been used in this study. Results. The iHDACs-induced increase in MDR1 mRNA levels is not followed by a subsequent increase in Pgp protein levels or activity in several pancreatic and colon carcinoma cell lines, suggesting a traslational control of Pgp in these cell lines. In addition, the MDR1 mRNA produced in these cell lines is shorter in its 5' end that the Pgp mRNA produced in cell lines expressing Pgp protein. The different size of the Pgp mRNA is due to the use of alternative promoters. We also demonstrate that these promoters are differentially regulated by TSA. The translational blockade of Pgp mRNA in the pancreatic carcinoma cell lines could be related to alterations in the 5' end of the MDR1 mRNA in the Pgp protein expressing cell lines. In addition, we demonstrate that the ABCB1 nested gene RUNDC3B expression although upregulated by TSA is independent of the ABCB1 alternative promoter used. Conclusions. The results show that the increase in MDR1 mRNA expression after iHDACs treatment is clinically irrelevant since this mRNA does not render an active Pgp protein, at least in colon and pancreatic cancer cell lines. Furthermore, we have demonstrated that TSA in fact, differentially regulates both ABCB1 promoters, downregulating the upstream promoter that is responsible for active P-glycoprotein expression. These results suggest that iHDACs such as TSA may in fact potentiate the effects of antitumoral drugs that are substrates of Pgp. Finally, we have also demonstrate that TSA upregulates RUNDC3B mRNA independently of the ABCB1 promoter in use.This work was supported by Spanish Health Ministry grants FIS-PI050969 and FIS-PI080901 and FIBElx-06/09 and 07/14 to MS, and by Caja Murcia to TMB

Diffuse leptomeningeal glioneuronal tumour: where to biopsy? Case report and literature review.

PURPOSE: Diffuse leptomeningeal glioneuronal tumour (DLGT) is an infrequent entity. Diagnosis is made with biopsy but with so few cases described management, prognosis remains undefined. There are not currently any articles regarding most effective place to biopsy. METHODS: Current literature review and introduction of the case of a 3-year-old male presenting at the emergency room with irritability, vomiting and nuchal rigidity. A head CT was made showing tetraventricular enlargement and a posterior fossa cyst. RESULTS: Patient underwent urgent ventriculoperitoneal shunting surgery with complete symptomp resolution. Brain MRI showed diffuse leptomeningeal enhancing, predominantly in basal cisterns, and multiple cystic-solid lesions along the neural axis. After ruling other conditions, a biopsy among intraoperative samples, was obtained of a PET positive gadolinium spinal enhancing lesion at D8 level, with the final diagnosis of DLGT. CONCLUSION: DLGT can present as acute hydrocephalus. Biopsy stablishes the diagnosis but the place to take the sample can be difficult to select. Our experience suggests that PET-CT and intraoperative biopsy analysis can improve the effectivity of a representative sample