https://hdl.handle.net/10481/68141 2026-04-20T02:08:35Z https://hdl.handle.net/10481/69348 To coagulate or not to coagulate. That is the question de Mota Dengra, Elena; Hidalgo Manchado, Laura; Pérez Villares, José Miguel Decision-making is a complex issue especially in severe multi-pathological patients, whose processes interact with each other and with other treatments, making it impossible to follow regular protocols. In this case, the patient suffered from a cranioencephalic traumatism and a pulmonary thromboembolism. In the case of pulmonary thromboembolism, an anticoagulation prophylaxis is recommended while in the former it is contraindicated. Moreover, complications may appear, apparently related or not to main diseases. In these situations, it is essential to establish priorities and have a vast knowledge of patients' pathologies and the possible complications that may emerge. Firstly, our aim was the resolution of the brain trauma due to the mortal risk of brain herniation. Straightaway we relieved the severe subsequent complications of the main processes, namely, refractory intracranial hypertension, acute respiratory failure and thrombocytopenia. Once the patient was stable, pulmonary thromboembolism treatment could be set and followed after clinical improvement and discharge. https://hdl.handle.net/10481/69347 Severe acute pancreatitis of gallblader origin with sequels: pancreatic necrosis, pseudocyst and splenic vein thrombosis Mochón Benguigui, Sol; Navarro Freire, Francisco 40 year old male with ulcerative gastritis, cholelithiasis and severe acute pancreatitis with long-term hospital stay, requiring tracheal intubation and tracheotomy. Chyloperitoneum, cholelithiasis and pancreatic pseudocyst with a 90% necrosis without active bleeding were found during surgery. Cholecistectomy, washing and intracavitary necrosectomy, stitching of the small vascular intracystic stump, Roux-en-Y cystojejunostomy, draining of the chyloperitoneum and of the cavity were performed. Chylous ascites persisted and was treated with octeotride, diuretics and fat-free diet supplemented with medium-chain trigliyerides, proteins and vitamins. However, sequels remained including: recurrent acute pancreatitis, splenic vein thrombosis and left portal hypertension. As the patient was not diabetic and the platelet count was 140000 cells/mm3 splenectomy or expectant monitoring were posed as possible choices. The interest of the case is rooted in the diagnosis, treatment and evolution of the chylous ascites, 90% pancreatic necrosis, pseudocyst and splenic vein thrombosis, as well as the repeated admissions. https://hdl.handle.net/10481/69346 Basics of isolation and cultivation of chondrocytes according to good laboratory practice Muñoz Olmedo, Jose Manuel; Harvanová, Denisa; Špaková, Timea; Rosocha, Ján; Lacko, Marek; Bačenková, Darina; Amrichová, Judita OBJECTIVES: The objective of the present study was to determine if chondrocytes isolated from human cartilage of five elderly patients (mean age 63) with osteoarthritis (stage 3) maintain their proliferation and chondrogenic potential. Isolation and cultivation of chondrocytes was performed according to good laboratory practice (GLP) standards. Methods: Chondrocytes were isolated from cartilage biopsy by enzymatic digestion. Cultivation of cells was performed in a controlled environment (cleanroom). Phenotype characterization of chondrocytes was achieved by flow cytometry analysis. Results: Three weeks after cultivation, polygonal structures typical of chondrocytes were observed, but spindle/fibroblast like morphology was also detected in cultured cells. Flow cytometric analysis showed that chondrocytes were positive for CD44 (98.35% ± 0.50), CD90 (97.15% ± 0.13) after first passage (P1) and the cells were negative for hematopoietic marker CD45 (0.21% ± 0.11). ConclusionS: Human articular chondrocytes obtained from five elderly patients with osteoarthritis maintained a chondrocyte phenotype and could be potentially used for autologous implantation. We have standardized the conditions for cultivation according to GLP standards to minimize the risk of in vitro cell contamination. https://hdl.handle.net/10481/69345 Primary cilium, cortactine, and Kv10.1 dynamics under confocal microscopy Sánchez, José Manuel; Carretero, Irene; González, Jorge; Urrego, Diana; Stühmer, Walter; Pardo, Luis A. OBJECTIVES: It is believed that tumor cells expressing Kv10.1 channel acquire selective advantages that allow them to maintain chronic proliferation. The ciliary disassembly is a prerequisite to enter into the cell cycle, therefore our objective was to find correlations between primary cilium disassembly and the Kv10.1 channel through the Kv10.1 cortactin binding site. Methods: Wild type Kv10.1, as well as the mutant form lacking the binding site to the cytoskeleton protein cortactin, were overexpressed in RPE-TERT cell line. The effect on ciliary disassembly of such overexpression forms was imaging using confocal microscopy. Results: Kv10.1 wild type overexpression induced disassembly of the primary cilium and alteration of the cytoskeleton, which corresponded to overproduction of filament structures positive to α-tubulin acetylated in the cytoplasm. Overexpression of mutated Kv10.1 did not show this pattern, nor the untreated cells. ConclusionS: This data suggests that Kv10.1 channel could be interfering with the cilium formation, and thus influences the cell cycle in a way that cortactine binding site is not involved. That should be furtherly studied because due to its possible implications in cancer. https://hdl.handle.net/10481/69344 BRCAness and breast cancer: BRCAness y cáncer de mama Betancor-Fernández, Isabel Breast cancer constitutes the neoplasm of highest incidence, prevalence and mortality among women in western countries. Around 25-30% of these tumors are estrogen receptor (ER), progesterone receptor (PR) and HER2 negative. These tumors are called triple negative breast cancer (TNBC). TNBCs frequently appear in young patients. Familial aggregation is common. They show a high histological grade and larger lymph node loco-regional affectation when diagnosed. In addition, they recur and progress prematurely after the standard combined treatment. A part of these TNBCs -up to 50% according to some authors- show specific phenotypical and molecular features that allow them to be considered BRCAness positive. BRCAness encompasses a combination of changes, of varied nature. These changes point towards a deficit in homology-directed repair (HDR). These TNBC types are generated and ensure their survival through the accumulation of unrepaired damage that increases genome instability. Recently it has been demonstrated that these tumors could be susceptible to specific treatments (alkylating agents, PARP inhibitors) with better response and survival rates. This leads to a greater accumulation of unrepaired damage that directs tumor cells towards apoptosis. Several techniques have been proposed to study BRCAness. Among them: conventional immunohistochemistry techniques with specific markers; pyrosequencing to determine the methylation degree of the BRCA1 promoter; multiplex ligationdependant probe amplification (MLPA), comparative genome hybridization, quantitative real-time PCR and, more recently, miRNAs and tumor circulating DNA study through digital droplet PCR (ddPCR).