AAV vectors transduce hepatocytes in vivo as efficiently in cirrhotic as in healthy rat livers.

In liver cirrhosis, abnormal liver architecture impairs efficient transduction of hepatocytes with large viral vectors such as adenoviruses. Here we evaluated the ability of adeno-associated virus (AAV) vectors, small viral vectors, to transduce normal and cirrhotic rat livers. Using AAV serotype-1 (AAV1) encoding luciferase (AAV1Luc) we analyzed luciferase expression with a CCD camera. AAV1Luc was injected through the hepatic artery (intra-arterial (IA)), the portal vein (intra-portal (IP)), directly into the liver (intra-hepatic (IH)) or infused into the biliary tree (intra-biliar). We found that AAV1Luc allows long-term and constant luciferase expression in rat livers. Interestingly, IP administration leads to higher expression levels in healthy than in cirrhotic livers, whereas the opposite occurs when using IA injection. IH administration leads to similar transgene expression in cirrhotic and healthy rats, whereas intra-biliar infusion is the least effective route. After 70% partial hepatectomy, luciferase expression decreased in the regenerating liver, suggesting lack of efficient integration of AAV1 DNA into the host genome. AAV1Luc transduced mainly the liver but also the testes and spleen. Within the liver, transgene expression was found mainly in hepatocytes. Using a liver-specific promoter, transgene expression was detected in hepatocytes but not in other organs. Our results indicate that AAVs are convenient vectors for the treatment of liver cirrhosis.

Detección y aislamiento de células inmaduras neuronales a partir de líneas celulares de tumores del sistema nervioso / Detection and isolation of immature neural cells from tumoral cell lines of the nervous system

Introducción: Hoy se sugiere que la teoria de carcinogénesis establecida en las últimas décadas, según la cual el cáncer se produce por el acúmulo de mutaciones en proto-oncogenes y genes supresores de tumores podría cuestionarse, a favor de una teoría más completa que incluya la participación de las denominadas células stem tumorales. Estas células poseerían la capacidad de iniciar y mantener el tumor, además de su propia capacidad de diferenciación y autorrenovación. Objetivo: Determinar el fenotipo de marcadores de diferenciación y la posible existencia de células inmaduras (tal vez células stem) en las líneas celulares de tumores del sistema nervioso. Material y métodos: Se estudió el nivel de expressión de los genes CD133, nestina, Musashi-1, FAS, NCAM1 y GFAP en 27 líneas celulares de tumores del sistema nervioso mediante RT-PCR semicuantitativa y citometría. La línea de neuroblastoma IMR-32 fue sometida a separación celular mediante SdFFF. Resultados: Hemos podido separar diferentes subclones o células en diferentes estadios de diferenciación en la línea de neuroblastoma IMR-32. La correcta tipificación de estas líneas celulares podría ser relevante para establecer tratamientos quimioterápicos o de terapia génica, específicamente dirigidos contra los subclones celulares más inmaduros, que podrían corresponder (o no) a células stem tumorales (AU)

Introduction: Nowadays it is suggested that the theory of carcinogenesis established along the last decades, according to which cancer is produced by the accumulation of mutations in proto-oncogenes and tumor suppressor genes might be questioned in favor of a more complete theory that includes the participation of the so called tumor stem cells. These cells would represent those in charge of initiating and maintaining the tumor; with their own capacity of differentiation and autorenewal. Objective: To determine the phenotype of differentiation markers and the possible existence of immature cells (maybe stem cells) in cell lines of tumors of the nervous system. Material and methods: The level of expression of CD133, nestine, Musashi-1, FAS, NCAM1 and GFAP genes was studied in 27 cell lines of tumors of the nervous system by semiquantitative RT-PCR and cytometry. The neuroblastoma cell line IMR-32 was subjected to cell separation by SdFFF. Results:We could separate different subclones or cells in different stages of differentiation in the neuroblastoma cell line IMR-32. The correct description of these cells might be relevant to set up chemoterapeutic or gene therapy treatments specifically targeted against the most immature subclones, that might correspond (or not) to tumor stem cells (AU)

Glycogen synthase kinase-3 is involved in the regulation of the cell cycle in cerebellar granule cells.

Recent studies have demonstrated that neuronal reentry in the cell cycle and specifically the expression of the transcription factor E2F-1, constitutes a pathway that may be involved in neuronal apoptosis after serum and potassium withdrawal. Other enzymes such as glycogen synthase kinase-3beta (GSK-3beta) are also involved in this apoptotic stimulus, and thus in the process of neuronal cell death. Primary cerebellar granule cells (CGNs) were used in this study to determine whether pharmacological inhibition of GSK-3beta is involved in neuronal modulation of the cell cycle, and specifically in the regulation of E2F-1 and retinoblastoma protein (Rb). CGNs showed a dramatic increase in GSK-3beta activity after 2h of serum and potassium deprivation. Immunoblot and activity assays revealed that lithium and SB415286 inhibit fully the activation of GSK-3beta and attenuate the expression of cyclin D, cyclin E, pRb phosphorylation and the transcription factor E2F-1. These data were confirmed using AR-014418, a selective GSK-3beta inhibitor that prevents the expression of cell-cycle proteins. Our data indicate that GSK-3beta inhibition regulates, in part, the cell cycle in CGNs by inhibiting Rb phosphorylation and thus inhibiting E2F-1 activity. However, the selective inhibition of GSK-3beta with AR-A014418 had not effect on cell viability or apoptosis mediated by S/K withdrawal. Furthermore, our results suggest that selective GSK-3beta inhibition is not sufficient to protect against apoptosis in this S/K withdrawal model, indicating that Li(+) and SB415286 neuroprotective effects are mediated by the inhibition of additional targets to GSK3beta. Therefore, there is a connection between cell cycle and GSK-3beta activation and that these, along with other mechanisms, are involved in the molecular paths leading to the apoptotic process of rat CGNs triggered by S/K withdrawal.

Complicaciones perianestésicas en una paciente con osteomalacia no conocida secundaria a cirugía bariátrica previa / Perianesthetic complications in a woman with undiagnosed osteomalacia secondary to earlier gastric bypass surgery

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Inhibition of phosphatidylcholine synthesis is associated with excitotoxic cell death in cerebellar granule cell cultures.

Glucose deprivation (GD) enhances the sensitivity of cerebellar granule cells to die by excitotoxicity. Neither 70 min of GD, a treatment that depletes cell energy resources, nor exposure to 20 microM glutamate (GLU) for 30 min, induce significant cell death in cultures of cerebellar granule cells. However, the combined treatment with GLU and GD induces choline (Cho) release before excitotoxic cell death. We investigated whether the neurotoxic effect of this treatment is related with inhibition of phosphatidylcholine (PC) synthesis. We found that exposure to GLU for 30 min, to GD for 70 min, and to the combination of both, inhibited PC synthesis at the end of treatment by 71%, 92% and 91%, respectively. The inhibition of PC synthesis was accompanied by a decrease in the incorporation of [(3)H]Cho into phosphocholine and by an increase of the intracellular content of free [(3)H]Cho, indicating that these treatments inhibit the synthesis of PC by inhibiting choline kinase activity. However, only the combined treatment with GLU and GD induced a prolonged inhibition of PC synthesis that extended after the end of treatment. These results show that excitotoxic death is associated with sustained inhibition of PC synthesis and suggest that this effect of the combined treatment with GLU and GD on PC synthesis is produced by an action on an enzymatic step downstream of choline kinase activity.

Hallazgo de quiste hidatídico intraarterial en el curso de toracotomía diagnóstica / An intra-arterial hydatic cyst found during diagnostic thoracotomy

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[Reduction of blood transfusion need with aprotinin in orthopedic surgery. Spanish Study Group on the Use of Aprotinin in Hip Arthroplasty (GEEEAAC)]. / Reducción de las necesidades transfusionales con aprotinina en cirugía ortopédica. Grupo Español de Estudio del Empleo de Aprotinina en Artroplastia de Cadera (GEEEAAC).

Aprotinin is a protease inhibitor of interest for its antifibrinolytic effect of reducing perioperative bleeding in certain types of surgery, with wide use in heart surgery, liver transplantation and vascular surgery. The application of aprotinin during orthopedic surgery has recently been suggested. Such use is controversial, as there is lack of consensus as to the type of patient for whom aprotinin administration would be indicated, the surgical procedure during which it would be most effective (hip or knee arthroplasty, spinal arthrodesis, major tumor or septic surgery), the doses to administer, its safety and its real efficacy for conserving homologous blood. That is to say, there is no agreement as to the cost/benefit relation of aprotinin for the various types of orthopedic surgery. This critical review of the literature leads to the conclusion that aprotinin is a promising drug for use in orthopedic surgery, given that published studies have established the benefit in blood product savings and decreased blood loss during surgery.

Reducción de las necesidades transfusionales con aprotinina en cirugía ortopédica / Reduced transfusional requirements with aprotinin in orthopedic surgery

La aprotinina es un inhibidor de las proteasas que tiene interés en la actualidad en su calidad de antifibrinolítico para disminuir el sangrado perioperatorio en determinados tipos de cirugía, y su uso está admitido ampliamente en cirugía cardíaca, en el trasplante hepático y en cirugía vascular.Recientemente se ha propuesto su empleo en cirugía ortopédica. Se trata de una indicación controvertida por la falta de unanimidad en el tipo de paciente en el que la aprotinina estaría indicada, en el procedimiento quirúrgico en el que se conseguiría una mayor efectividad (artroplastia de cadera, artroplastia de rodilla, artrodesis raquídea, cirugía mayor tumoral o séptica), en las dosis que se deben administrar, en la seguridad de su empleo y en la eficacia real en el ahorro de sangre homóloga. Es decir, no hay acuerdo en cuanto al rendimiento de la relación coste/beneficio del fármaco en los diferentes procedimientos de cirugía ortopédica.En esta revisión se hace un estudio crítico de las publicaciones al respecto, concluyendo finalmente que se trata de un fármaco prometedor en cirugía ortopédica, dado que en los estudios publicados se ha obtenido un beneficio en relación con el ahorro de hemoderivados y con la disminución de sangrado perioperatorio (AU)

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[Heart arrest in cemented hip arthroplasty]. / Paro cardíaco en artroplastia de cadera cementada.

Hip arthroplasty is a common surgical intervention in our hospital practice, involving high perioperative risk related to patients age and multiple concomitant diseases. Hemodynamic complications described vary from slight hypotension during surgery to heart failure and sudden death, particularly if the operation involves a cemented femoral component. Because of the type of patients undergoing such operations (elderly patients, with osteoporosis and scarce cardiopulmonary reserve), the unclear origin of complications and the lack of consensus on what constitutes adequate monitoring during surgery, hip arthroplasty is problematic for the specialists involved. We report on five deaths during cemented hip arthroplasty; after reviewing the case history and autopsy report of one, we believe the events leading to death were triggered by massive pulmonary embolism.

Análisis de la expresión del pepsinógeno C en metástasis ganglionares de carcinoma de mama / Analysis of pepsinogen C expression in lymph node metastases of breast cancer

En este estudio analizamos en los tumores primarios y en ganglios axilares metastásicos de 30 mujeres con cáncer de mama, mediante análisis inmunohistoquímico, la expresión tumoral del pepsinógeno C, una proteína normalmente expresada por la mucosa gástrica. De los tumores mamarios primarios, 16 (53,3 por ciento) mostraron una tinción inmunohistoquímica positiva para el pepsinógeno C, mientras que 17 (56,6 por ciento) lo hicieron en sus ganglios linfáticos tumorales. Además existió una relación significativamente positiva entre la expresión tumoral de pepsinógeno C en los tumores primarios y los ganglios metastásicos (p < 0,002). Sin embargo, sólo la expresión tumoral de esa proteína en los tumores primarios alcanzó significación estadística (p < 0,05) como factor pronóstico de evolución favorable para predecir la supervivencia de las pacientes. (AU)

Apolipoprotein D expression in metastasic lymph nodes of breast cancer.

BACKGROUND: Apolipoprotein D is a glycoprotein of the human plasma whose functional role remains unclear. On the other hand, this protein is also produced by breast carcinomas and is positively associated with a favorable outcome of patients. However, none study has focused on metastasic lesions. AIM: To analyze apolipoprotein D expression in breast cancer patients and their synchronous metastasic axillary lymph nodes. METHODS: We analyzed by immunohistochemical assay both, the tumoral expression of apolipoprotein D in primary tumors and in their synchronous metastasic axillary lymph nodes of 30 node-positive breast cancer patients. RESULTS: Of the primary tumors, 28 (93.3%) showed a positive immunostaining for apolipoprotein D, although there was wide variability immunostaining values. On the other hand, 16 (53.3%) patients showed a positive immunostaining for the protein in their tumoral lymph nodes. In addition, there was a significant positive relationship between the tumoral expression of apolipoprotein D in primary tumors and metastasic lymph nodes (P < 0.05). However, only immunostaining values of the protein in primary tumors achieve statistical signification (P < 0.05) as prognostic factor of favorable evolution to predict overall survival from patients. CONCLUSIONS: Apolipoprotein D is also expressed in metastasic lymph nodes of breast carcinomas, but with a different pattern of immunostaining and less clinical significance than in primary tumors.

[The utility of digital infiltration of mepivacaine and ketorolac in postoperative analgesia of the unilateral hallux valgus]. / Utilidad de la infiltración digital con mepivacaína y ketorolaco en la analgesia postoperatoria del hallux valgus unilateral.

OBJECTIVES: To determine whether locally injected ketorolac provides analgesia additional to that of mepivacaine, and also to prevent, diminish or delay the peripheral hypersensitivity response of postoperative pain. PATIENTS AND METHODS: Prospective, randomized, double-blind study of 72 patients scheduled for surgery to correct unilateral hallux valgus. Group 1 (n = 24) received median infiltration at the first metatarsus of 5 ml of 2% mepivacaine and 1 ml (30 mg) of ketorolac. Group 2 (n = 21) received local infiltration of 5 ml of 2% mepivacaine and 1 ml of saline solution. Group 3, the control group (n = 27) received the same solution as did group 2, plus 30 mg of ketorolac intravenously. The postoperative analgesia prescribed was 10 mg of ketorolac orally every 8 hours. Pain was measured on a visual analog scale (VAS) 0, 1, 4, 8 and 24 hours after surgery. Time elapsed until the appearance of pain, number of ketorolac pills consumed and overall patient satisfaction were recorded. RESULTS: There were no differences in anthropometric characteristics. Time until pain appeared was significantly longer in group 1 than in groups 2 and 3 (14.66 +/- 7.19, 5.90 +/- 2.27 and 8.70 +/- 5.02 hours, respectively). The VAS scores were significantly lower in group 1 after the fourth postoperative hour. Analgesic consumption was significantly lower in group 1. CONCLUSIONS: Infiltration of 30 mg of ketorolac along with mepivacaine delays the appearance of postoperative pain and diminishes it in the first 24 hours after surgery to correct hallux valgus, in comparison with infiltration of mepivacaine alone plus intravenous ketorolac.

A case of cardiac hydatidosis.

We present the case of a female patient with a diagnosis of hydatidosis located in the heart. Although echinococcosis is endemic to our country, very few cases of cardiac hydatidosis are normally reported. In our patient, the hydatid cyst was located in the septum and in the right ventricular cavity; it presented other unusual features, such as the fact that it was located exclusively in the heart, that it first manifested as anaphylactic shock of unknown origin and that it required immediate surgical treatment because of severe haemodynamic compromise.

[Secondary ureterectomy in an ectopic ureteral opening into the seminal vesicle]. / Ureterectomía secundaria en la desembocadura ureteral ectópica en vesícula seminal.

The ectopic ureter opening to the seminal vesicle is uncommon in the male and even less in a duplex kidney. It commonly presents as recurrent urinary infection with pelviperineal pain. Treatment is by heminephrectomy with total ureterectomy and removal of the seminal vesicle if it is cystic. Herein we describe a 70-year-old patient who had previously undergone a heminephrectomy and partial ureterectomy due to an ectopic ureter opening to the seminal vesicle of the ureter of the upper pelvis of the left kidney. The patient was submitted to a second ureterectomy procedure due to pyoureter in the ureteral stump. The main features of this pathological condition are described and the surgical approach is discussed.

[Parotid metastasis of renal adenocarcinoma: report of a case]. / Metástasis parotidea de adenocarcinoma renal: a propósito de un caso.

Renal adenocarcinomas are tumours which sometimes tend to remain clinically silent, and initially become evident through distant metastasis. This paper presents the case of a 54 year-old patient whose initial clinical evidence was parotid metastasis from kidney clear cells adenocarcinoma. The patient had a metastasis and renal tumour exeresis with application of supplementary chemotherapy and immunotherapy, presenting early pulmonary metastasis which remains currently unchangeable. Some comments apropos of distant metastasis, its prognosis and regression in renal tumours are made.