RESUMEN
BACKGROUND: Chronic urticaria can be the initial clinical presentation of a number of different diseases. The objective of the present study was to report the associated diseases during a ten-year clinical-laboratory follow-up in patients with an initial diagnosis of chronic spontaneous urticaria (CSU) of unknown cause. METHODS: A prospective, longitudinal cohort study with a ten-year clinical-laboratory follow-up was conducted. Patients with a history of urticarial plaques of over six weeks presenting as the only clinical symptom were selected. Individuals with other clinical conditions, urticaria of known causes or chronic physical urticaria were excluded. The following tests were initially performed: haemogram, urine type I, stool parasite exam and sedimentation rate. The following exams were ordered during follow-up: PPD; urine culture; serology tests; antithyroid and antinuclear antibodies, rheumatoid factor, lupus anticoagulant; thyroid hormones; serum immunoglobulin; paranasal sinus and thorax radiographs; testing for BK and Helicobacter pylori; and prick tests. RESULTS: Infections were diagnosed in 29% of patients (syphilis, parasitosis, H. pylori, urinary infection, tuberculosis, hepatitis B and C); autoimmune diseases in 21% (thyroiditis, rheumatoid arthritis and antiphospholipid antibody syndrome); primary immunodeficiencies in 4% (IgA and IgG2 deficiencies); and chronic myeloid leukaemia in 1%. At ten-years of follow-up, the urticaria diagnosis was CSU of unknown cause in 45% of the cases. CONCLUSION: This ten-year clinical-laboratory follow-up of 100 individuals with chronic urticaria as the initial diagnosis revealed the presence of associated diseases in over half of the cases. The most prevalent diseases were infections and autoimmune diseases besides primary immunodeficiencies and blood diseases.
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Enfermedades Autoinmunes/complicaciones , Enfermedades Transmisibles/complicaciones , Disgammaglobulinemia/complicaciones , Urticaria , Adulto , Enfermedades Autoinmunes/diagnóstico , Enfermedades Autoinmunes/epidemiología , Enfermedad Crónica , Enfermedades Transmisibles/diagnóstico , Enfermedades Transmisibles/epidemiología , Disgammaglobulinemia/diagnóstico , Disgammaglobulinemia/epidemiología , Femenino , Estudios de Seguimiento , Humanos , Estudios Longitudinales , Masculino , Prevalencia , Estudios Prospectivos , Pruebas Cutáneas , Urticaria/diagnóstico , Urticaria/epidemiología , Urticaria/inmunologíaAsunto(s)
Neoplasias Colorrectales/patología , Colorantes/farmacocinética , Sistemas de Liberación de Medicamentos/métodos , Verde de Indocianina/farmacocinética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/secundario , Imagen Molecular/métodos , Colorantes/administración & dosificación , Fluorescencia , Humanos , Verde de Indocianina/administración & dosificación , Neoplasias Hepáticas/patologíaRESUMEN
The present work is part of a preclinical in vitro study to assess the efficacy of BNCT applied to liver or lung coloncarcinoma metastases and to limb osteosarcoma. Adherent growing cell lines can be irradiated as adherent to the culture flasks or as cell suspensions, differences in radio-sensitivity of the two modalities of radiation exposure have been investigated. Dose related cell survival and cell cycle perturbation results evidenced that the radiosensitivity of adherent cells is higher than that of the suspended ones.
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Terapia por Captura de Neutrón de Boro , Adhesión Celular/efectos de la radiación , Rayos gamma , Neoplasias/radioterapia , Neutrones , Animales , Ciclo Celular/efectos de la radiación , Línea Celular Tumoral , Neoplasias/patología , RatasRESUMEN
Ischemia reperfusion injury (IRI) is a major field of study in small bowel transplantation because of its implications regarding intestinal immunity. In this study, we have introduced some variations to the described models of IRI in pigs to make possible a complete isolation of the small bowel for IRI studies. In swine, two anatomical barriers make impossible a complete isolation of the small bowel at the origin of superior mesenteric artery (SMA) and vein (SMV): the main colic vessels, which originate distally to form SMA and SMV, and the blood supply of the distal portion of the duodenum and the cephalic part of the pancreas. In a group of Large White pigs (n = 5), we have performed a complete isolation of the small bowel, including sub-total colectomy and pancreaticoduodenectomy. Both SMA and SMV were isolated at the origin from the aorta and at the junction of the splenic vein, respectively. Intestinal continuity was restored with duodenojejunal anastomosis and with ileotransverse colon anastomosis. One pig died on postoperative day 5 from intestinal occlusion due to adhesions. The remaining four pigs were killed on postoperative day 7 after an uneventful postoperative course. No complications were found at autopsy. In swine, resection of part of the pancreas and duodenum and removal of the large bowel does not affect short-term survival, allowing a full isolation of the entire small bowel mimicking the transplantation procedure. Thus, this model appears to be attractive for IRI studies in the field of intestinal transplantation.
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Intestino Delgado/trasplante , Arteria Mesentérica Superior/cirugía , Daño por Reperfusión/prevención & control , Animales , Modelos Animales de Enfermedad , Intestino Delgado/irrigación sanguínea , Sus scrofa , Porcinos , Trasplante AutólogoRESUMEN
AIM: The standard to treat liver tumors is a resection. When the future liver remnant (FLRV) is below 30% (healthy livers) or 40% (cirrhotic livers or previous chemotherapy), surgery carries the risk of severe complications. Portal vein embolization (PVE) gained a worldwide diffusion as a tool to augment the FLRV. Cell therapies are recent players at the frontiers of medicine. This study presents a clinical experience to evaluate the synergistic effect of combined PVE and autologous CD133+ cells coadministration. METHODS: Sixteen patients have been enrolled in the study up today. Inclusion criteria were: primary or metastatic liver malignancy with a FLRV<30% or 40%. A baseline volumetric CT-scan was obtained. CD34+ were mobilized to the blood stream by G-CSF administration and collected by immunomagnetic separation. Simultaneously with PVE, cells were administered to the non occluded liver segments. Follow-up CT scans were taken at 30th post treatment day. RESULTS: The patients (N.=6) showed an increased volume gain (Mann-Whitney test P<0.001, two sided) compared to a set of cases whose treatment was PVE only (N.=10). DISCUSSION: The use of autologous stem cells as an augmenter of liver regeneration has a clinical potential to improve the resectability of liver tumors.
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Antígenos CD/análisis , Embolización Terapéutica , Glicoproteínas/análisis , Neoplasias Hepáticas/cirugía , Regeneración Hepática , Péptidos/análisis , Trasplante de Células Madre de Sangre Periférica/métodos , Vena Porta , Antígeno AC133 , Antígenos CD34/análisis , Factor Estimulante de Colonias de Granulocitos/farmacología , Movilización de Célula Madre Hematopoyética , Leucaféresis , Hígado/diagnóstico por imagen , Hígado/patología , Hígado/cirugía , Neoplasias Hepáticas/terapia , Tamaño de los Órganos , Tomografía Computarizada Espiral , Trasplante AutólogoRESUMEN
Thanks to their immunonodulatory properties, multipotent mesenchymal stromal cells (MSCs) are a promising strategy for preventing/reducing the risk of graft rejection after hematopoietic cell and solid organ transplantation. We have previously demonstrated that porcine MSCs (pMSCs) can be isolated from bone marrow and display similar morphology and differentiative capacity as compared to human MSC (hMSCs). In this study, we investigated the in vitro immunomodulatory properties (namely the ability to suppress lymphocyte proliferation in response to phytohemagglutinin and the cytokine production in the culture supernatants) of pMSCs from six Large White 6-month old piglets. Similarly to hMSCs, pMSCs reduced the phytohemagglutinin-induced lymphocyte proliferation. High levels of IL-6 were found in culture supernatants, whereas IL-10 and TGF-ß were not detectable. In conclusion, ex vivo expanded pMSCs share selected biological/functional properties with hMSCs. pMSCs may be used in in vivo models to investigate novel approaches of prevention of graft rejection in solid organ transplantation.
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Células de la Médula Ósea/inmunología , Células Madre Mesenquimatosas/inmunología , Células Madre Multipotentes/inmunología , Animales , Proliferación Celular , Células Cultivadas , Humanos , Interleucina-10/inmunología , Interleucina-6/inmunología , Linfocitos/citología , Linfocitos/inmunología , Porcinos , Factor de Crecimiento Transformador beta/inmunologíaRESUMEN
(10)B molecular compounds suitable for Boron Neutron Capture Therapy (BNCT) are tagged with a Gd(III) paramagnetic ion. The newly synthesized molecule, Gd-BPA, is investigated as contrast agent in Magnetic Resonance Imaging (MRI) with the final aim of mapping the boron distribution in tissues. Preliminary Nuclear Magnetic Resonance (NMR) measurements, which include (1)H and (10)B relaxometry in animal tissues, proton relaxivity of the paramagnetic Gd-BPA molecule in water and its absorption in tumoral living cells, are reported.
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Terapia por Captura de Neutrón de Boro , Boro , Gadolinio , Isótopos , Animales , Imagen por Resonancia Magnética , Espectroscopía de Resonancia Magnética , Neoplasias Experimentales/metabolismo , Protones , RatasRESUMEN
Boron neutron capture therapy (BNCT) is a binary radiotherapy based on thermal-neutron irradiation of cells enriched with (10)B, which produces α particles and (7)Li ions of short range and high biological effectiveness. The selective uptake of boron by tumor cells is a crucial issue for BNCT, and studies of boron uptake and washout associated with cell survival studies can be of great help in developing clinical applications. In this work, boron uptake and washout were characterized both in vitro for the DHDK12TRb (DHD) rat colon carcinoma cell line and in vivo using rats bearing liver metastases from DHD cells. Despite a remarkable uptake, a large boron release was observed after removal of the boron-enriched medium from in vitro cell cultures. However, analysis of boron washout after rat liver perfusion in vivo did not show a significant boron release, suggesting that organ perfusion does not limit the therapeutic effectiveness of the treatment. The survival of boron-loaded cells exposed to thermal neutrons was also assessed; the results indicated that the removal of extracellular boron does not limit treatment effectiveness if adequate amounts of boron are delivered and if the cells are kept at low temperature. Cell survival was also investigated theoretically using a mechanistic model/Monte Carlo code originally developed for radiation-induced chromosome aberrations and extended here to cell death; good agreement between simulation outcomes and experimental data was obtained.
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Apoptosis/efectos de la radiación , Terapia por Captura de Neutrón de Boro/métodos , Neoplasias del Colon/metabolismo , Neoplasias del Colon/radioterapia , Animales , Boro/farmacocinética , Boro/uso terapéutico , Línea Celular Tumoral , Isótopos/farmacocinética , Isótopos/uso terapéutico , Masculino , Tasa de Depuración Metabólica , Radiofármacos/farmacocinética , Radiofármacos/uso terapéutico , Ratas , Distribución Tisular , Resultado del TratamientoRESUMEN
Transvaginal recovery of the kidney has recently been reported, in a donor who had previously undergone a hysterectomy, as a less-invasive approach to perform laparoscopic live-donor nephrectomy. Also, robotic-assisted laparoscopic kidney donation was suggested to enhance the surgeon's skills during renal dissection and to facilitate, in a different setting, the closure of the vaginal wall after a colpotomy. We report here the technique used for the first case of robotic-assisted laparoscopic live-donor nephrectomy with transvaginal extraction of the graft in a patient with the uterus in place. The procedure was carried out by a multidisciplinary team, including a gynecologist. Total operative time was 215 min with a robotic time of 95 min. Warm ischemia time was 3 min and 15 s. The kidney was pre-entrapped in a bag and extracted transvaginally. There was no intra- or postoperative complication. No infection was seen in the donor or in the recipient. The donor did not require postoperative analgesia and was discharged from the hospital 24 h after surgery. Our initial experience with the combination of robotic surgery and transvaginal extraction of the donated kidney appears to open a new opportunity to further minimize the trauma to selected donors.
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Laparoscopía/métodos , Donadores Vivos , Nefrectomía/métodos , Robótica , Recolección de Tejidos y Órganos/métodos , Adulto , Colpotomía , Femenino , Humanos , Trasplante de Riñón/métodos , Masculino , Persona de Mediana Edad , VaginaRESUMEN
Mesenchymal stem cells (MSC) are multipotent cells that differentiate into various mature cell lineages. MSC show immunomodulatory effects by inhibiting T-cell proliferation. We evaluated the effect of the infusion of MSC in rats experimental kidney transplantation. Sprague-Dawley transgenic rats (SD) able to express the green fluorescent protein (EGFP) were used as MSC donors. Syngeneic (Lewis to Lewis, n = 10) and allogeneic (Fischer to Lewis, n = 10) kidney transplantations were performed after bilateral nephrectomy. Five transplanted rats who received syngeneic grafts, were treated with 3 x 10(6) MSC (Gr B), while the other 5 did not received MSC (Gr A). Five rats with allogenic grafts received 3 x 10(6) MSC (Gr C) and another 5 did not receive MSC (Gr D). The MSC were infused directly into the renal artery of the graft. No immunosuppressive therapy was provided. The animals were killed after 7 days. Biochemical analysis for renal function, histological (Banff criteria) and immunohistological analysis (ED1+ and CD8+) were performed on treated animals. MSC improved kidney function in Gr B and D vs Gr A and C. The tubular damage appeared to be less severe among Gr B and Gr D with respect to Gr A and C (P < .01). Vasculitis was more accentuated in Gr A and C (P < .01). MSCs reduced the inflammatory infiltrate; in Gr B and D, the number of ED1+ cells was lower than in Gr A and C (P < .005), which was also observed for CD8+ cells (P < .05). Our study demonstrated that the infusion of MSC attenuated histological damage from acute rejection by reducing the cellular infiltration.
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Rechazo de Injerto/prevención & control , Trasplante de Riñón/inmunología , Trasplante de Células Madre Mesenquimatosas , Animales , Animales Modificados Genéticamente , Técnicas de Cultivo de Célula , Diuresis , Proteínas Fluorescentes Verdes/genética , Masculino , Células Madre Mesenquimatosas/citología , Proteinuria , Ratas , Ratas Endogámicas F344 , Ratas Endogámicas Lew , Ratas Sprague-Dawley , Trasplante IsogénicoRESUMEN
Immunomodulating cell therapy represents a new perspective for the control of cellular immune responses that determine the occurrence of acute rejection (ACR) in allo-transplantation. Mesenchymal stem cells (MSC) demonstrate immunoregulatory effects by inactivating T-cell components that regulate tissue damage in transplantation models. The presumed mechanism of action is recruitment of cells by a cytokine network. The purpose of this study was to test which route of administration (intra-arterial vs intravenous) was the most effective route to achieve immunomodulating effects in experimental rat kidney transplantation. Transgenic Sprague-Dawley rats (SD) expressing the enhanced green fluorescent protein (EGFP) at the somatic level were used as MSC donors: Allogeneic Fischer to Lewis grafts (n = 4 per group) were performed in rats after bilateral nephrectomy. In Gr B, 3 x 10(6) MSCs were infused into the renal graft artery, whereas in Gr C, they were infused into the tail vein. The untreated Gr A were a control group. No immunosuppressive therapy was administered. The animals were sacrificed at day 7 postoperatively. Biochemical analysis for renal function, histological (Banff criteria) and immunohistological (anti-EGFP-Immunoglobulin) analysis were performed on the transplanted animals. In Gr B, functional recovery was more rapid (creatinine: Gr B vs Gr C, P < .05). The inflammatory infiltrate in the graft was less in Gr B vs Gr C, with preservation of tubules, arteries, and glomeruli (P < .01). Intra-arterial infusion of MSCs was more effective to control ACR.
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Trasplante de Riñón/fisiología , Trasplante de Células Madre Mesenquimatosas/métodos , Animales , Técnicas de Cultivo de Célula , Citometría de Flujo , Proteínas Fluorescentes Verdes/genética , Infusiones Intraarteriales , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/fisiología , Ratas , Ratas Endogámicas Lew , Ratas Sprague-Dawley , Ratas Transgénicas , Ratas Wistar , Trasplante HeterotópicoRESUMEN
Pharmacological aspecific immunosuppression, despite being widely used in solid organ transplantation recipients, is unable to completely prevent allograft rejection. It promotes the occurrence of sometimes life-threatening infections. Due to their immunosuppressive and anti- inflammatory properties, there is great interest in the therapeutic use of bone marrow (BM)-derived mesenchymal stromal cells (MSC). Large animal models play a crucial role to investigate the biological and functional properties of MSCs as novel cellular therapy. In the current study we sought to isolate expand ex vivo, and phenotypically characterize MSC derived from BM of 4 Large White 6-month-old piglets. Porcine MSC (pMSC) were characterized for their in vitro differentiation capacity. pMSC were successfully isolated from all BM samples. They showed spindle-shaped morphology and a stable doubling time on culture. They were positive for CD90, CD29, CD105, and negative for CD45 and CD11b. Furthermore, they differentiated, upon specific in vitro conditions toward adipogenic and osteogenic lineages. The optimization of methods for the isolation and characterization of pMSC may be useful to elucidate their biological and functional properties. The anatomy and physiology of the pig, which is similar to humans, make this animal model more attractive than small animals to test the safety and efficacy of MSC in the context of solid organ transplantation.
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Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/citología , Animales , Antígenos CD/análisis , Células de la Médula Ósea/citología , Diferenciación Celular , División Celular , Medios de Cultivo , Trasplante de Células Madre Hematopoyéticas , Humanos , Tolerancia Inmunológica , Porcinos , Tolerancia al TrasplanteRESUMEN
BACKGROUND: Patients with atopic dermatitis frequently present recurrent infections by pyogenic bacteria or by intracellular microorganisms, suggesting an immune disorder. OBJECTIVE: Laboratorial investigation of phagocyte activity and chemotactic response by neutrophilic polymorphonuclear and mononuclear phagocytes in the peripheral blood of patients with atopic dermatitis from moderate to severe. METHODS: Through a transversal study, patients with atopic dermatitis from moderate to severe were selected. The neutrophilic and mononuclear phagocytes were separated and the phagocytic ingestion of zymosan particles was analysed, in addition to migration distance to the bacterial lipopolysaccharide chemotactic factor, comparing the results to the values obtained from healthy individuals within the same age group. RESULTS: Nineteen patients were selected, 11 female and 8 male. The mean age was 6.47 years (+/-4.65). Among the 19 patients studied, 14 (73.68%) presented a reduction in the neutrophilic and mononuclear phagocyte activity, with two (1.53%) patients presenting a reduction in the activity of both phagocytes. CONCLUSION: Our results demonstrated a reduction in chemotactic response and phagocytic activity by neutrophilic and/or mononuclear phagocytes in the majority of patients with atopic dermatitis from moderate to severe. Our results were coherent with the clinical data concerning the higher incidence of infections by pyogenic bacteria and fungi in patients with atopic dermatitis, which are microorganisms that require defence by the phagocytes researched in the present study.
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Quimiotaxis de Leucocito/inmunología , Dermatitis Atópica/inmunología , Fagocitos/inmunología , Fagocitosis/inmunología , Adolescente , Adulto , Edad de Inicio , Alérgenos/inmunología , Infecciones Bacterianas/complicaciones , Infecciones Bacterianas/inmunología , Niño , Preescolar , Dermatitis Atópica/complicaciones , Dermatitis Atópica/diagnóstico , Dermatitis Atópica/epidemiología , Femenino , Humanos , Hipersensibilidad/complicaciones , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Recuento de Leucocitos , Lipopolisacáridos/inmunología , Macrófagos/citología , Macrófagos/inmunología , Masculino , Micosis/complicaciones , Micosis/inmunología , Neutrófilos/citología , Neutrófilos/inmunología , Fagocitos/citología , Pruebas Cutáneas , Adulto Joven , Zimosan/inmunologíaRESUMEN
To test the possibility to apply boron neutron capture therapy (BNCT) to lung tumors, some rats are planned to be irradiated in the thermal column of the TRIGA reactor of the University of Pavia. Before the irradiation, lung metastases will be induced in BDIX rats, which will be subsequently infused with boronophenylalanine (BPA). During the irradiation, the rats will be positioned in a box designed to shield the whole animal except the thorax area. In order to optimize the irradiation set-up and to design a suitable shielding box, a set of calculations were performed with the MCNP Monte Carlo transport code. A rat model was constructed using the MCNP geometry capabilities and was positioned in a box with walls filled with lithium carbonate. A window was opened in front of the lung region. Different shapes of the holder and of the window were tested and analyzed in terms of the dose distribution obtained in the lungs and of the dose absorbed by the radiosensitive organs in the rat. The best configuration of the holder ensures an almost uniform thermal neutron flux inside the lungs (Phi(max)/Phi(min)=1.5), an irradiation time about 10 min long, to deliver at least 40 Gy(w) to the tumor, a mean lung dose of 5.9+/-0.4 Gy(w), and doses absorbed by all the other healthy tissues below the tolerance limits.
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Terapia por Captura de Neutrón de Boro/instrumentación , Terapia por Captura de Neutrón de Boro/estadística & datos numéricos , Neoplasias Pulmonares/radioterapia , Reactores Nucleares , Planificación de la Radioterapia Asistida por Computador/estadística & datos numéricos , Animales , Italia , Neoplasias Pulmonares/secundario , Modelos Animales , Método de Montecarlo , Fantasmas de Imagen , Protección Radiológica/instrumentación , Ratas , Efectividad Biológica RelativaRESUMEN
Almost eight years ago, in December 2001, we performed for the first time in the world thermal neutron irradiation on an isolated liver of a patient. The organ was affected by diffuse metastases of a colon carcinoma and had been previously loaded with a (10)B compound. In July 2003, the same procedure was applied again on a patient for the treatment of unresectable and incurable hepatic metastases of a carcinoma of the rectum. Both patients are dead at present. Now we can analyze in depth the clinical history of these patients and evaluate the effectiveness of this therapy. From this exciting experience we learned much, and we also found out about complications till then unknown, which need to be studied and addressed experimentally. Unfortunately we can base our conclusions just on the experience we had with these two patients. We could have been much more detailed and firm in our statements if the number of clinical cases was larger. The BNCT Pavia project has been suspended, but it is more than likely to resume in a short time. Good findings were many. The procedure is feasible; the original concept of complete immersion of the diseased liver in a homogeneous neutron field proved effective and winning. The tumor masses resulted completely necrotic and unknown metastases too appeared radically treated; healthy hepatic tissue was preserved from both morphological and functional points of view; no symptoms of cirrhosis appeared even four years after treatment. For the long term surviving patient, quality of life was excellent. Other findings require to be tackled in depth. The "post-irradiation syndrome" we observed in both patients, with identical symptoms and biochemical derangements, creates a dramatic--even though totally reversible--clinical condition, that is the probable cause of death for our second patient, suffering from cardiomyopathy, 33 days after treatment. For the first patient, recurrences were a late yet fatal complication, for which even a further surgical revision was ineffective. We offer some hypotheses about their origin and possible prevention.
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Terapia por Captura de Neutrón de Boro/métodos , Neoplasias Hepáticas/radioterapia , Neoplasias Hepáticas/secundario , Adulto , Compuestos de Boro/administración & dosificación , Compuestos de Boro/uso terapéutico , Terapia por Captura de Neutrón de Boro/efectos adversos , Cardiomiopatía Dilatada/etiología , Quimioterapia del Cáncer por Perfusión Regional/métodos , Neoplasias del Colon , Circulación Extracorporea , Humanos , Neoplasias Hepáticas/cirugía , Masculino , Persona de Mediana Edad , Necrosis , Fenilalanina/administración & dosificación , Fenilalanina/análogos & derivados , Fenilalanina/uso terapéutico , Traumatismos por Radiación/etiología , Fármacos Sensibilizantes a Radiaciones/administración & dosificación , Fármacos Sensibilizantes a Radiaciones/uso terapéutico , Neoplasias del Recto , Tomografía Computarizada por Rayos XRESUMEN
Osteosarcoma is the most common non-hematologic primary cancer type that develops in bone. Current osteosarcoma treatments combine multiagent chemotherapy with extensive surgical resection, which in some cases makes necessary the amputation of the entire limb. Nevertheless its infiltrative growth leads to a high incidence of local and distant recurrences that reduce the percentage of cured patients to less than 60%. These poor data required to set up a new therapeutic approach aimed to restrict the surgical removal meanwhile performing a radical treatment. Boron neutron capture therapy (BNCT), a particular radiotherapy based on the nuclear capture and fission reactions by atoms of (10)B, when irradiated with thermal neutrons, could be a valid alternative or integrative option in case of osteosarcoma management, thanks to its peculiarity in selectively destroying neoplastic cells without damaging normal tissues. Aim of the present work is to investigate the feasibility of employing BNCT to treat the limb osteosarcoma. Boronophenylalanine (BPA) is used to carry (10)B inside the neoplastic cells. As a first step the endocellular BPA uptake is tested in vitro on the UMR-106 osteosarcoma cell line. The results show an adequate accumulation capability. For the in vivo experiments, an animal tumor model is developed in Sprague-Dawley rats by means of an intrafemoral injection of UMR-106 cells at the condyle site. The absolute amounts of boron loading and the tumor to normal tissue (10)B ratio are evaluated 2 h after the i.v. administration of BPA. The boron uptake by the neoplastic tissue is almost twice the normal one. However, higher values of boron concentration in tumor are requested before upholding BNCT as a valid therapeutic option in the treatment of osteosarcoma.
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Compuestos de Boro/farmacocinética , Compuestos de Boro/uso terapéutico , Terapia por Captura de Neutrón de Boro/métodos , Osteosarcoma/metabolismo , Osteosarcoma/radioterapia , Fenilalanina/análogos & derivados , Fármacos Sensibilizantes a Radiaciones/farmacocinética , Fármacos Sensibilizantes a Radiaciones/uso terapéutico , Animales , Transporte Biológico Activo , Neoplasias Óseas/metabolismo , Neoplasias Óseas/radioterapia , Línea Celular Tumoral , Supervivencia Celular/efectos de la radiación , Técnicas In Vitro , Isótopos/farmacocinética , Isótopos/uso terapéutico , Masculino , Fenilalanina/farmacocinética , Fenilalanina/uso terapéutico , Ratas , Ratas Sprague-Dawley , Distribución TisularRESUMEN
INTRODUCTION: The shortage of organs in the last 20 years is stimulating the development of new strategies to expand the pool of donors. The harvesting of a graft from non-heart-beating donors (NHBDs) has been successfully proposed for kidney and liver transplantation. To our knowledge, no studies are available for small bowel transplantation using NHBDs. In an experimental setting of small bowel transplantation, we studied the feasibility of using intestinal grafts retrieved from NHBDs. MATERIALS AND METHODS: Twenty five Large White piglets underwent total orthotopic small bowel transplantation and were randomly divided as follow: NHBD group (n = 15) received grafts from NHBDs; heart-beating donor (HBD) group (n = 10) received grafts from HBDs. The NHBD pigs were sacrificed inducing the cardiac arrest by a lethal potassium injection. After 20 minutes (no touch period = warm ischemia), they underwent cardiac massage, laparotomy, and aorta cannulation for flushing and cooling the abdominal organs. In HBDs, the cardiac arrest was induced at the time of organ cold perfusion. In both groups, immunosuppression was based on tacrolimus oral monotherapy. The animals were observed for 30 days. The graft absorptive function was studied at day 30 using the D-xylose absorption test. Histological investigation included HE (Hematoxilin and Eosin) microscopical analysis and immunohistological staining. RESULTS: Animals in the NHBD group died due to infection (n = 3), acute cellular rejection (n = 2), technical complications (n = 2), and intestinal failure (n = 8). In the HBD group, all animals but two were alive at the end of the study. The D-xylose absorption was significantly lower among the NHBD compared with the HBD group (P < .05). CONCLUSIONS: This study confirmed that intestinal mucosa is sensitive to ischemic injury. When the intestinal graft is harvested from NHBDs, the infectious-related mortality was higher and the absorptive function lower. Histological examination confirmed a higher grade of ischemic injury in the NHBD grafts that correlated with the clinical data. Therefore, this experimental study suggested that non-heart-beating donation may not be indicated for small bowel transplantation.
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Intestino Delgado/trasplante , Animales , Peso Corporal , Muerte Encefálica , Supervivencia de Injerto , Paro Cardíaco/inducido químicamente , Inmunosupresores/uso terapéutico , Mucosa Intestinal/patología , Isquemia/etiología , Donadores Vivos , Modelos Animales , Potasio/toxicidad , Porcinos , Factores de Tiempo , Donantes de Tejidos , Trasplante Homólogo , Xilosa/uso terapéuticoRESUMEN
Malononitrilamide 715 (FK778) is a new class of immunosuppressant, derived from the active metabolite of leflunomide A77 1726. We investigated the efficacy of two different immunosuppressive induction protocols with tacrolimus plus FK778 followed by FK778 monotherapy. In a swine model of small bowel transplantation, we observed three groups, divided by different therapy regimens: group 1 (n = 5): no immunosuppressant (control group); group 2 (n = 10): oral tacrolimus (from postoperative day [POD] 0 to 30) and FK778 (from POD 0 to 60); group 3 (n = 8): oral tacrolimus, as group 2, and FK778 (from POD 7 to POD 60). Median survival was 11, 60, and 21 days in groups 1, 2, and 3, respectively. In group 1 all animals died of acute rejection; in group 2 the causes of death were technical complication (n = 1) and sepsis (n = 1); in group 3, one animal died from obstruction, two from pneumonia, one from peritonitis, one from sepsis. Group 2 accounted for 0.5 infection episode/animal versus 0.62 in group 3 (P < .05). Acute rejection was absent or mild in 66% and 75% of group 3 and 2 biopsies, respectively (P < .05). The D-xylose absorption curves from groups 2 and 3 were similar to those of the nontransplanted healthy animals. In conclusion, FK778 monotherapy after a consistent induction period with tacrolimus combined immunosuppression is able to extend survival and preserve optimal absorptive capacity of the small bowel allograft in our pig model. The association of tacrolimus and FK778 from day 1, compared to the delayed administration of FK778 from day 7, results in a significant reduction of infections and postoperative complications.
Asunto(s)
Alquinos/uso terapéutico , Intestino Delgado/trasplante , Isoxazoles/uso terapéutico , Nitrilos/uso terapéutico , Trasplante Homólogo/fisiología , Adyuvantes Inmunológicos/uso terapéutico , Animales , Infecciones Bacterianas/mortalidad , Causas de Muerte , Modelos Animales , Complicaciones Posoperatorias/clasificación , Complicaciones Posoperatorias/mortalidad , Sepsis/mortalidad , Análisis de Supervivencia , Porcinos , Trasplante Homólogo/mortalidadRESUMEN
The main goals for a successful small bowel transplantation (SBTx) are the control of acute rejection and maintenance of the mucosal barrier, which plays a key role in preventing bacterial translocation and preserving absorptive capacity. According to recent evidence that sustaining enteral nutrition (EN) as rehabilitative therapy improves the integrity of the mucosal barrier after SBTx, we studied the trophic effect of a new elemental enteral solution whose proteinic supply is represented by oligomeric-aminoacidic chains. In a swine SBTx model we studied three groups, divided by the different postoperative feeding: group 1 (n = 5): standard swine chow, group 2 (n = 5): polymeric enteral solution, group 3 (n = 5): elemental enteral solution (Peptamen, Nestlè Corp). All animals were immunosuppressed with a tacrolimus/FK778 combined oral therapy. The nutritional indices evaluated were: body weight, episodes of diarrhea, D-xylose absorption test, and histopatological and villi morphometric analysis. Three pigs died before the end of the study, two in group 1 (pneumonia and sepsis), one in group 2 (pneumonia). Mean days of diarrhea were 15, 10, and 3 in groups 1, 2, and 3, respectively (P < .05). The final/starting weight ratio was 1.08 for group 3 and 0.92 for group 2 (P < .05); the D-xylose curves showed a statistically significant difference for group 3 versus the groups 2 and 1 (P < .05), as well as for the villi height (P < .01) and width (P < .05). In conclusion, elemental enteral solution, with its basic protein supply, does not require a very complex enzymatic system to be metabolized. Thus, it may contribute to a faster recovery of the mucosal barrier and to limit the hypercatabolic state.
Asunto(s)
Nutrición Enteral , Mucosa Intestinal/fisiología , Intestino Delgado/trasplante , Microvellosidades/fisiología , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Absorción Intestinal , Modelos Animales , Neumonía , Complicaciones Posoperatorias/clasificación , Sepsis , Porcinos , Trasplante Homólogo , XilosaRESUMEN
AIM: The liver as a solid graft has a known immunological privilege. Its tolerogenic property has been demonstrated in rodents. In humans the onset of chronic rejection and the severity of such complication is less frequent after liver transplantation compared to other organs. The underlying events whose effect is graft acceptance instead of rejection should be further investigated. Their control could open new ways to decrease the need for long-term immunosuppression after transplantation of other organs. Aim of this study is to evaluate a model of liver transplantation in swine as a preliminary step for immunological studies. METHODS: Ten outbred Landrace/Large White mismatched swine underwent to liver transplantation with a simple passive portocaval jugular bypass. The onset of rejection was monitored daily by liver function test. After death or sacrifice the liver parenchyma was studied to evaluate tissue damage and inflammatory infiltrate. RESULTS: The postoperative liver function showed a critical period for organ rejection about postoperative day 5. The animals that survived longer were sacrificed with a normal biochemical hepatic function. However, histology consistently showed a pattern of mild rejection in a still preserved architecture. CONCLUSIONS: The evidence of a prolonged liver function in a rejecting model of liver transplantation makes this model suitable for studies of tolerance induction.