The teaching of Radiology in other university studies.

After the implementation of the European Space for Higher Education, the contents of the Radiology and Physical Medicine Area that were taught in the Medicine Degree have also been incorporated into the new degrees of Dentistry, Nursing, Physiotherapy, Podiatry, and, to a lesser extent, Pharmacy, Occupational Therapy, Logopedia, and Biomedical Engineering As a whole, the basic concepts of radiology and radiological protection are taught in Murcia in 5 different degrees with a total of 52.5 ECTS credits, participating in the training of 1219 students each academic year. This incorporation in the new degrees has tripled the number of subjects in which undergraduate teaching is taught, and doubled both the number of ECTS credits and the number of undergraduate students to whom it directs its training work. Thus, given the possible creation of new university degrees in the near future (Diagnostic Imaging and Radiotherapy Technicians), it would be necessary to involve a greater number of accredited professionals, from different specialties, and to optimize teaching resources (bibliography, material teacher, clinical cases, etc.,) for its usefulness in the different subjects that share similar contents.

La enseñanza de la Radiología en otros estudios universitarios / The teaching of Radiology in other university studies

Tras la implantación del Espacio Europeo de Formación Superior, los contenidos del Área de Radiología y Medicina Física que se impartían tradicionalmente en la Licenciatura de Medicina se han incorporado también a los nuevos grados de Odontología, Enfermería, Fisioterapia, Podología y, en menor medida, Farmacia, Terapia Ocupacional, Logopedia, e Ingeniería Biomédica. En su conjunto, los conceptos básicos de radiología y protección radiológica se imparten en Murcia en 5 grados diferentes con un total de 52,5 créditos ECTS, participando en la formación de 1219 alumnos cada curso académico. Esta incorporación en los nuevos grados ha triplicado el número de asignaturas en las que se imparte docencia pregrado, y duplicado tanto el número de créditos ECTS como el número de alumnos de pregrado a los que dirige su labor de formación. Así, ante la posible creación de nuevos grados universitarios en un futuro próximo (Imagen para el Diagnóstico y Técnico en Radioterapia) sería necesaria la implicación de un mayor número de profesionales acreditados, de diferentes especialidades, y que optimicen los recursos docentes (bibliografía, material docente, casos clínicos, etc.) para su utilidad en las diferentes asignaturas que comparten contenidos similares.(AU)

After the implementation of the European Space for Higher Education, the contents of the Radiology and Physical Medicine Area that were taught in the Medicine Degree have also been incorporated into the new degrees of Dentistry, Nursing, Physiotherapy, Podiatry, and, to a lesser extent, Pharmacy, Occupational Therapy, Logopedia, and Biomedical Engineering As a whole, the basic concepts of radiology and radiological protection are taught in Murcia in 5 different degrees with a total of 52.5 ECTS credits, participating in the training of 1,219 students each academic year. This incorporation in the new degrees has tripled the number of subjects in which undergraduate teaching is taught, and doubled both the number of ECTS credits and the number of undergraduate students to whom it directs its training work. Thus, given the possible creation of new university degrees in the near future (Diagnostic Imaging and Radiotherapy Technicians), it would be necessary to involve a greater number of accredited professionals, from different specialties, and to optimize teaching resources (bibliography, material teacher, clinical cases, etc.) for its usefulness in the different subjects that share similar contents.(AU)

A new method to quantify ß-antithrombin glycoform in plasma reveals increased levels during the acute stroke event.

INTRODUCTION: ß-antithrombin, the minor antithrombin glycoform in plasma, is probably the major thrombin inhibitor in vivo because of its high heparin affinity. The levels and variability of this glycoform in general population and its relevance in thromboembolic diseases is unknown since there is no specific method to measure this glycoform in clinical samples. METHODS: Plasma and recombinant α- and ß-antithrombins were purified by heparin affinity chromatography. An anti-FXa chromogenic method in presence of pentassacharide was used with two NaCl concentrations (15mM and 1.1M). This method was applied to plasma samples from 97 healthy subjects and 117 consecutive patients with ischemic cerebrovascular disease during the acute event and one year later. SERPINC1 sequencing was done in cases with antithrombin deficiency. RESULTS: High salt concentrations specifically restricted the pentassacharide-induced activation of antithrombin to the ß glycoform. ß-antithrombin displayed a normal distribution in the general population (89.5%-103.5%), with no significant variations according to age or sex. In patients, whole antithrombin values remained within the normal range. Only five cases had antithrombin deficiency during the thrombotic event, one carrying the L99F mutation in SERPINC1. Interestingly, both ß-antithrombin and the ß/whole antithrombin ratio were significantly higher in patients during the acute event but normalized after one year. CONCLUSIONS: We have developed a rapid, simple, sensitive and specific method to quantify ß-antithrombin activity using 1µL of plasma. ß-antithrombin significantly increases in patients with ischemic cerebrovascular disease during the acute event, probably by its release from the vasculature.

An atypical IgM class platelet cold agglutinin induces GPVI-dependent aggregation of human platelets.

Platelet cold agglutinins (PCA) cause pseudothrombocytopenia, spurious thrombocytopenia due to ex vivo platelet clumping, complicating clinical diagnosis, but mechanisms and consequences of PCA are not well defined. Here, we characterised an atypical immunoglobulin (Ig)M PCA in a 37-year-old woman with lifelong bleeding and chronic moderate thrombocytopenia, that induces activation and aggregation of autologous or allogeneic platelets via interaction with platelet glycoprotein (GP)VI. Patient temperature-dependent pseudothrombocytopenia was EDTA-independent, but was prevented by integrin αIIbß3 blockade. Unstimulated patient platelets revealed elevated levels of bound IgM, increased expression of activation markers (P-selectin and CD63), low GPVI levels and abnormally high thromboxane (TX)A2 production. Patient serum induced temperature- and αIIbß3-dependent decrease of platelet count in allogeneic donor citrated platelet-rich plasma (PRP), but not in PRP from Glanzmann's thrombasthenia or afibrinogenaemia patients. In allogeneic platelets, patient plasma induced shape change, P-selectin and CD63 expression, (14)C-serotonin release, and TXA2 production. Activation was not inhibited by aspirin, cangrelor or blocking anti-Fc receptor (FcγRIIA) antibody, but was abrogated by inhibitors of Src and Syk, and by a soluble GPVI-Fc fusion protein. GPVI-deficient platelets were not activated by patient plasma. These data provide the first evidence for an IgM PCA causing platelet activation/aggregation via GPVI. The PCA activity persisted over a five-year follow-up period, supporting a causative role in patient chronic thrombocytopenia and bleeding.

Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations.

BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). Recombinant mutants were generated and purified. Glycoform heterogeneity and conformational sensitivity were studied with electrophoresis, proteomic analysis, and glycomic analysis. Heparin affinity was evaluated from intrinsic fluorescence. Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. RESULTS AND CONCLUSIONS: Pleiotropic mutants, except for that with the M315K mutation, which affects a non-exposed residue, showed two glycoforms. Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. Our results suggest the C-sheet of antithrombin as a new region that is relevant for proper maturation of the N-glycans. Therefore, pleiotropic mutations lead to glycosylation defects that are responsible for the reduced heparin affinity.

The value of 18F-FDG PET/CT for assessing the response to neoadjuvant therapy in locally advanced rectal cancer.

PURPOSE: Neoadjuvant radiochemotherapy (RCT) is an accepted treatment for locally advanced rectal cancer (LARC) that improves surgical outcomes. If a pathological complete response is achieved, conservative surgery can be considered. The objective of our study was to assess the reliability of (18)F-FDG PET/CT for evaluating the response to neoadjuvant RCT in LARC. METHODS: We prospectively studied 41 patients diagnosed with LARC and candidates for neoadjuvant RCT. PET/CT was performed before RCT and again 7 weeks later. A visual and semiquantitative analysis was carried out. The pathological response was classified according to the Mandard tumour regression grade (TRG). We analysed: (a) the relationship between TRG and the result of the posttreatment PET/CT scan, and (b) the correlation between the percentage of pathological response and the percentage decrease in SUVmax according to the response index (RI). RESULTS: The mean SUVmax of the rectal lesions at diagnosis was 13.6 and after RCT 3.96. The mean RI was 65.32 %. Sensitivity was 88.88 %, specificity 92.86 %, positive predictive value 96 %, negative predictive value 81 %. Of the 41 patients, 8 had TRG I (all negative PET/CT); 6 had TRG II (5 negative, 1 positive PET/CT); 16 had TRG III (13 positive, 3 negative PET/CT); 9 had TRG IV (all positive PET/CT); 2 had TRG V (all positive PET/CT). Of the 14 patients classified as responders (TRG I, II), 13 (92.86 %) had negative PET/CT. Of the 27 patients classified as nonresponders (TRG III-V), 24 (88.88 %) had positive PET/CT. Differences were statistically significant (p < 0.0001). The RI in responders was 79.9 % and in nonresponders was 60.3 %. Differences were statistically significant (p < 0.037). CONCLUSION: PET/CT is a reliable technique for assessing response to neoadjuvant RCT in LARC, with a view to considering more conservative surgical treatment. The combination of the visual and semiquantitative analysis increases the diagnostic validity of PET/CT.

Type II antithrombin deficiency caused by a large in-frame insertion: structural, functional and pathological relevance.

BACKGROUND: The metastable native conformation of serpins is required for their protease inhibition mechanism, but also renders them vulnerable to missense mutations that promote protein misfolding with pathological consequences. OBJECTIVE: To characterize the first antithrombin deficiency caused by a large in-frame insertion. PATIENTS/METHODS: Functional, biochemical and molecular analysis of the proband and relatives was performed. Recombinant antithrombin was expressed in HEK-EBNA cells. Plasma and recombinant antithrombins were purified and sequenced by Edman degradation. The stability was evaluated by calorimetry. Reactive centre loop (RCL) exposure was determined by thrombin cleavage. Mutant antithrombin was crystallized as a dimer with latent plasma antithrombin. RESULTS: The patient, with a spontaneous pulmonary embolism, belongs to a family with significant thrombotic history. We identified a complex heterozygous in-frame insertion of 24 bp in SERPINC1, affecting strand 3 of ß-sheet A, a region highly conserved in serpins. Surprisingly, the insertion resulted in a type II antithrombin deficiency with heparin binding defect. The mutant antithrombin, with a molecular weight of 59 kDa, had a proteolytic cleavage at W49 but maintained the N-terminal disulphide bonds, and was conformationally sensitive. The variant was non-inhibitory. Analysis of the crystal structure of the hyperstable recombinant protein showed that the inserted sequence annealed into ß-sheet A as the fourth strand, and maintained a native RCL. CONCLUSIONS: This is the first case of a large in frame-insertion that allows correct folding, glycosylation, and secretion of a serpin, resulting in a conformationally sensitive non-inhibitory variant, which acquires a hyperstable conformation with a native RCL.

In silico discovery of a compound with nanomolar affinity to antithrombin causing partial activation and increased heparin affinity.

The medical and socioeconomic relevance of thromboembolic disorders promotes an ongoing effort to develop new anticoagulants. Heparin is widely used as activator of antithrombin but incurs side effects. We screened a large database in silico to find alternative molecules and predicted d-myo-inositol 3,4,5,6-tetrakisphosphate (TMI) to strongly interact with antithrombin. Isothermal titration calorimetry confirmed a TMI affinity of 45 nM, higher than the heparin affinity (273 nM). Functional studies, fluorescence analysis, and citrullination experiments revealed that TMI induced a partial activation of antithrombin that facilitated the interaction with heparin and low affinity heparins. TMI improved antithrombin inhibitory function of plasma from homozygous patients with antithrombin deficiency with a heparin binding defect and also in a model with endothelial cells. Our in silico screen identified a new, non-polysaccharide scaffold able to interact with the heparin binding domain of antithrombin. The functional consequences of this interaction were experimentally characterized and suggest potential anticoagulant therapeutic applications.

Heparin affinity of factor VIIa: implications on the physiological inhibition by antithrombin and clearance of recombinant factor VIIa.

Factor VIIa (FVIIa), a trypsin-like serine protease, plays an essential role in haemostasis by initiating the coagulation in complex with its cofactor, tissue factor (TF). The TF pathway inhibitor is the main physiological inhibitor of FVIIa-TF complex, but FVIIa can also be inhibited by antithrombin, although little is known about this process. Functional analyses by second order kinetic determination and identification of FVIIa-antithrombin complex by electrophoresis, evaluating the effect of different cofactors: pentasaccharide, low molecular weight heparin (LMWH) and unfractionated heparin (UFH), confirmed that any activation of antithrombin significantly enhanced the inhibition of FVIIa. The analysis of the binding of FVIIa to heparin by surface plasmon resonance identified a high affinity interaction under physiologic conditions (K(D)=3.38 µM, with 0.15M of ionic strength) strongly dependent on Ca(2+) and ionic strength. This interaction was verified in cell models, indicating that FVIIa also binds to the surface of endothelial cells with similar requirements. Structural modeling suggests the presence of a potential exosite II in FVIIa. However, the binding of heparin did not display significant changes on both the intrinsic fluorescence and the associated functional consequences of FVIIa. These results indicate that FVIIa binds to exposed glycosaminglycans of the endothelium through an exosite II, structurally similar to that reported for thrombin and suggested for FIXa. This binding may favor its inhibition by antithrombin in the absence of TF, contributing to the physiological control of this protease. This process may also play an important role in the clearance of recombinant FVIIa administered to patients.

Validity of tests performed to diagnose acute abdominal pain in patients admitted at an emergency department.

OBJECTIVE: To determine the real importance of anamnesis, physical examination, and various tests in the assessment of acute abdominal pain. METHODS: A retrospective observational study with patients complaining of abdominal pain at the Emergency Department, Altiplano Health Area (Murcia) was performed. In our study we considered the following variables: socio-demographic data, history of previous surgery, symptoms, place and type of pain. Imaging tests were labeled as positive, negative, or inconclusive for assumed diagnoses, which were retrospectively assessed by an external radiologist who was unaware of the patient s final diagnosis. RESULTS: Our study includes 292 patients with a mean age of 45.49 years; 56.8% of these patients were women. Regarding the frequency of the different acute abdomen diagnoses, appendicitis was the main cause (approx. 25%), followed by cholecystitis (10%). We found a significant diagnostic correlation between pain location in the right hypochondrium (RHC) and a diagnosis with cholecystitis. This location was also significant for acute appendicitis (up to 74%). Regarding clinical signs, we only observed a significant correlation between fever and viscera perforation, and between Murphy s sign and cholecystitis. Sensitivity and specificity found in relation to the psoas sign were similar to those seen in other series, 16 and 95% respectively, and slightly lower than the Blumberg or rebound sign, which we found to be around 50 and 23%, respectively. CONCLUSIONS: a) Anamnesis and physical examination offer limited accuracy when assessing acute abdomen; b) ultrasound scans offer a low diagnostic agreement index for appendicitis; and c) laparoscopy may prove useful for diagnosis, and is also a possible treatment for acute abdominal pain despite its low diagnostic efficiency.

[Absence of chromosomic and genotoxic damage from the radiation dose administered in scintigraphic examinations]. / Ausencia de daño cromosómico y genotóxico inducido por la dosis de radiación administrada en las exploraciones gammagráficas.

INTRODUCTION: The cytokinesis-blocked (CB) micronucleus test (MN) on irradiated human lymphocytes is normally used to evaluate chromosomal and genotoxic damage produced by various physical and chemical agents. OBJECTIVE: Determine any possible genotoxic effect induced by the different types of ionizing radiation employed in medical diagnostic radiology and nuclear medicine. MATERIAL AND METHODS: The frequency of the MN appearance was determined in CB lymphocyte cultures of a total of 4 different groups of patients: (1) in 35 supposedly healthy volunteers to establish the MN spontaneous frequency in the medium; (2) in 9 volunteers to measure the in vitro dose-response curves in order to calculate the MN frequency following X-ray irradiation and gamma radiation; (3) in 25 patients in whom a specific diagnostic and/or therapeutic procedure employing diagnostic radiology techniques involving X-ray exposure was applied, and (4) in 26 patients in whom the diagnostic procedure in question involved nuclear medicine techniques (scintiscan). RESULTS: A lineal relationship was observed between the MN frequency and the dose of ionizing radiation administered in vitro, both in X-rays and gamma radiation. A significant increase in the MN is observed after radiation is given to patients during medical diagnostic radiology examinations when compared with the control values obtained from the same patients prior to being subjected to the radiological procedure (p < 0.01). No significant MN increase is observed following exposure to radiation involved in diagnostic examinations in patients studied in Nuclear Medicine. CONCLUSION: Ionizing radiation employed in complex medical diagnostic radiology examinations produces a significant increase in the MN appearance frequency and as such indicates both radiation induced chromosomal and genotoxic damage. However, the ionizing radiation used in diagnostic nuclear medicine examinations does not induce any significant increase in MN appearance frequency.

Actividad genotóxica de la terapia con I131 en pacientes tratados por carcinomas de tiroides / Genotoxic activity of the I 131 therapy in patients treated for thyroid carcinoma

- Introducción: El test de micronúcleos (MN) sobre linfocitos humanos irradiados con bloqueo citocinético (CB) se utiliza para valorar el daño cromosómico y genotóxico de diferentes agentes físicos y químicos.- Objetivo: Determinar un posible efecto genotóxico inducido por la terapia con I131 en pacientes con cáncer de tiroides y determinar la dosis equivalente corporal total (DECT) de radiación ionizante que supone dicho tratamiento.- Material y métodos: Se ha determinado la frecuencia de aparición de MN en cultivos de linfocitos CB en tres grupos de individuos diferentes: 1) en 35 voluntarios sanos para establecer la frecuencia espontánea de MN; 2) en 9 voluntarios supuestamente sanos para realizar las curvas dosisrespuesta "in vitro" con radiación gamma; y 3) en 25 pacientes que han recibido una dosis ablativa de I131 en el tratamiento de un carcinoma de tiroides. Se ha determinado el número de MN/500 células CB previo al tratamiento y tres días después de la administración de I131. La DECT de la terapia se ha calculado por el número de MN en linfocitos obtenido a los tres días de la administración de I131 comparada con la frecuencia de MN expuestas "in vitro" a radiación gamma que produciría una idéntica frecuencia de MN.- Resultados: Se ha obtenido una relación lineal entre la frecuencia de MN y la dosis de radiación ionizante administradas "in vitro" con radiación gamma. La frecuencia de MN tras el tratamiento con I131(8´89 MN/500CB) es significativamente mayor (p<0.01), duplicando la frecuencia espontánea (4´08/500MN) basal.- Conclusión: La terapia con I131 induce un incremento significativo del daño cromosómico en los pacientes irradiados por carcinoma de tiroides, equivalente a una dosis corporal total de 13 cGy durante los tres primeros días desde la administración terapéutica de I131 (AU)

[Selective sentinel node biopsy in melanoma using preoperative lymphoscintigraphy location and intraoperative detection gamma probe]. / Biopsia selectiva del ganglio centinela en el melanoma mediante localización con linfogammagrafía preoperatoria y sonda de detección gamma intraoperatoria.

BACKGROUND: The experience in detection of sentinel lymph node in melanoma using preoperative scintigraphy and intraoperative gamma probe is referred. PATIENTS AND METHODS: We studied 60 patients with stage I-II melanoma who underwent sentinel lymph node biopsy performed using 99m-Tc-labelled sulphur colloid as radioactive tracer. A preoperative scintigraphy was performed and intraoperative gamma probe was used to localize the sentinel node in all cases. Scintigraphy results, effectiveness of intraoperative detection (technical efficacy), pathological results, and follow-up have been studied. RESULTS: Preoperative detection was 98.3% and the mean basin detected was 1.17. There were multiple basins especially when melanomas were on the trunk. Technical efficacy was 98.4% and intraoperative detection was more difficult in parotid gland region. HMB-45 immunohistochemical staining was essential in pathological studies, in whom 10% were positives. Lymphadenectomy could be avoided in 90% of the patients. Recurrences were not detected during follow-up and metastases were found only in non biopsied cases. Sentinel node biopsy morbidity was significative lesser than that of lymphadenectomy. CONCLUSIONS: Preoperative scintigraphy and intraoperative gamma probe use to localize sentinel node in melanoma have a high efficacy. They can reveal multiple basins and they allow a more selective surgical approach and a minimal dissection.