RESUMEN
PURPOSE: Candida biofilm infections are frequently linked to the use of biomaterials and are of clinical significance because they are commonly resistant to antifungals. Clioquinol is an antiseptic drug and is effective against multidrug-resistant Candida. We investigated the effect of clioquinol and two other 8-hydroxyquinoline derivatives on Candida biofilm. METHODOLOGY: The ability to inhibit biofilm formation, inhibit preformed biofilm and remove established biofilms was evaluated using in vitro assays on microtitre plates. The action of clioquinol on biofilm in intrauterine devices (IUDs) was also investigated, describing the first protocol to quantify the inhibitory action of compounds on biofilms formed on IUDs. RESULTS: Clioquinol was found to be the most effective 8-hydroxyquinoline derivative among those tested. It prevented more than 90â% of biofilm formation, which can be attributed to blockade of hyphal development. Clioquinol also reduced the metabolic activity of sessile Candida but the susceptibility was lower compared to planktonic cells (0.031-0.5 µg ml-1 required to inhibit 50â% planktonic cells and 4-16 µg ml-1 to inhibit 50â% preformed biofilms). On the other hand, almost complete removal of biofilms was not achieved for the majority of the isolates. Candida spp. also showed the ability to form biofilm on copper IUD; clioquinol eradicated 80-100â% of these biofilms. CONCLUSION: Our results indicate a potential application in terms of biomaterials for 8-hydroxyquinoline derivatives. Clioquinol could be used as a coating to prevent morphological switching and thus prevent biofilm formation. Furthermore, clioquinol may have future applications in the treatment of Candida infections linked to the use of IUDs.
Asunto(s)
Antifúngicos/farmacología , Biopelículas/efectos de los fármacos , Candida/efectos de los fármacos , Candidiasis/prevención & control , Clioquinol/farmacología , Oxiquinolina/farmacología , Antifúngicos/química , Antifúngicos/uso terapéutico , Candida/fisiología , Candidiasis/tratamiento farmacológico , Candidiasis/etiología , Candidiasis/microbiología , Clioquinol/análogos & derivados , Clioquinol/química , Clioquinol/uso terapéutico , Cobre , Femenino , Humanos , Dispositivos Intrauterinos/efectos adversos , Dispositivos Intrauterinos/microbiología , Pruebas de Sensibilidad Microbiana , Oxiquinolina/análogos & derivados , Oxiquinolina/químicaRESUMEN
BACKGROUND: Lipiodol (iodized poppy seed oil) accumulates predominately in the tumor rather than in the liver tissue [1, 2]. Therefore, mixing anticancer drugs with Lipiodol may enhance the antitumor effect by increasing the local drug concentration. OBJECTIVE: In this pilot study, we made use of Lipiodol as a potential carrier of three promising antitumor metal complexes (tris(8-quinolato)gallium(III) (KP46), tetrachlorobis(indazole)ruthenate(III) (KP1019) and the hydrolysis product of KP1019, mer,trans-[RuCl3(H2O)(Hind)2]. METHODS: The stability of the drugs in Lipiodol and the release profile into the aqueous phase were examined independently by three different analytical techniques (high pressure liquid chromatography, HPLC; atom absorption spectroscopy, AAS; and electron spray ionization mass spectrometry, ESI-MS). RESULTS: The complexes were stable and remained in the Lipiodol emulsion over 3 days. In contrast to KP1019 and KP46, evaluation of Lipiodol emulsions of mer,trans-[RuCl3 (H2O) (Hind) 2] was not possible due to the insolubility of the compound in Lipiodol. KP1019 released rapidly into the aqueous phase in the first week and after 1 month it was not possible to detect the complex in the emulsion. KP46 showed a gradual release with the time resulting in the release of about 6.4 % of KP46 into the aqueous phase after 1 month of incubation. CONCLUSION: The initial results show that Lipiodol can be successfully employed as a carrier of anticancer Ru- or Ga-complexes. Furthermore, advantages can overcome the poor water solubility of the metal complexes, opening new perspectives for the use of Lipiodol emulsions in molecular imaging and cancer therapy as theragnostic agents.
Asunto(s)
Antineoplásicos/química , Aceite Etiodizado/química , Indazoles/química , Compuestos Organometálicos/química , Oxiquinolina/análogos & derivados , Cromatografía Líquida de Alta Presión , Emulsiones/química , Hidrólisis , Imagen Molecular , Estructura Molecular , Oxiquinolina/química , Proyectos Piloto , Compuestos de Rutenio , Solubilidad , Espectrometría de Masa por Ionización de Electrospray , Espectrofotometría AtómicaRESUMEN
BACKGROUND: Osteosarcoma is the most frequent primary malignant bone tumor. Although survival has distinctly increased due to neoadjuvant chemotherapy in the past, patients with metastatic disease and poor response to chemotherapy still have an adverse prognosis. Hence, development of new therapeutic strategies is still of utmost importance. METHODS: Anticancer activity of KP46 against osteosarcoma cell models was evaluated as single agent and in combination approaches with chemotherapeutics and Bcl-2 inhibitors using MTT assay. Underlying mechanisms were tested by cell cycle, apoptosis and autophagy assays. RESULTS: KP46 exerted exceptional anticancer activity at the nanomolar to low micromolar range, depending on the assay format, against all osteosarcoma cell models with minor but significant differences in IC50 values. KP46 treatment of osteosarcoma cells caused rapid loss of cell adhesion, weak cell cycle accumulation in S-phase and later signs of apoptotic cell death. Furthermore, already at sub-cytotoxic concentrations KP46 reduced the migratory potential of osteosarcoma cells and exerted synergistic effects with cisplatin, a standard osteosarcoma chemotherapeutic. Moreover, the gallium compound induced signs of autophagy in osteosarcoma cells. Accordingly, blockade of autophagy by chloroquine but also by the Bcl-2 inhibitor obatoclax increased the cytotoxic activity of KP46 treatment significantly, suggesting autophagy induction as a protective mechanism against KP46. CONCLUSION: Together, our results identify KP46 as a new promising agent to supplement standard chemotherapy and possible future targeted therapy in osteosarcoma.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Neoplasias Óseas/tratamiento farmacológico , Compuestos Organometálicos/farmacología , Osteosarcoma/tratamiento farmacológico , Oxiquinolina/análogos & derivados , Autofagia/efectos de los fármacos , Neoplasias Óseas/patología , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Sinergismo Farmacológico , Humanos , Indoles , Terapia Molecular Dirigida , Compuestos Organometálicos/administración & dosificación , Osteosarcoma/patología , Oxiquinolina/administración & dosificación , Oxiquinolina/farmacología , Pirroles/administración & dosificaciónRESUMEN
PURPOSE: Anticancer dendritic cell (DC) vaccines require the DCs to relocate to lymph nodes (LN) to trigger immune responses. However, these migration rates are typically very poor. Improving the targeting of ex vivo generated DCs to LNs might increase vaccine efficacy and reduce costs. We investigated DC migration in vivo in humans under different conditions. EXPERIMENTAL DESIGN: HLA-A*02:01 patients with melanoma were vaccinated with mature DCs loaded with tyrosinase and gp100 peptides together with keyhole limpet hemocyanin (NCT00243594). For this study, patients received an additional intradermal vaccination with (111)In-labeled mature DCs. The injection site was pretreated with nonloaded, activated DCs, TNFα, or Imiquimod; granulocyte macrophage colony-stimulating factor was coinjected or smaller numbers of DCs were injected. Migration was measured by scintigraphy and compared with an intrapatient control vaccination. In an ex vivo tissue model, we measured CCL21-directed migration of (19)F-labeled DCs over a period of up to 12 hours using (19)F MRI to supplement our patient data. RESULTS: Pretreatment of the injection site induced local inflammatory reactions but did not improve migration rates. Both in vitro and in vivo, reduction of cell numbers to 5 × 10(6) or less cells per injection improved migration. Furthermore, scintigraphy is insufficient to study migration of such small numbers of (111)In-labeled DCs in vivo. CONCLUSION: Reduction of cell density, not pretreatment of the injection site, is crucial for improved migration of DCs to LNs in vivo.
Asunto(s)
Vacunas contra el Cáncer/uso terapéutico , Células Dendríticas/inmunología , Melanoma/terapia , Adolescente , Adulto , Anciano , Vacunas contra el Cáncer/inmunología , Movimiento Celular , Células Dendríticas/citología , Células Dendríticas/trasplante , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Humanos , Ganglios Linfáticos/citología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/patología , Masculino , Melanoma/inmunología , Melanoma/patología , Persona de Mediana Edad , Estadificación de Neoplasias , Compuestos Organometálicos/química , Oxiquinolina/análogos & derivados , Oxiquinolina/química , Linfocitos T Citotóxicos/inmunologíaRESUMEN
Mixed ligand complexes of dioxouranium(VI) of the type [UO2(Q)(L)-2H2O] have been synthesized using 8-hydroxyquinoline (HQ) as a primary ligand and N- and/or O- donor amino acids (HL) such as L-lysine, L-aspartic acid and L-cysteine as secondary ligands. The metal complexes have been characterized on the basis of elemental analysis, electrical conductance, room temperature magnetic susceptibility measurements, spectral and thermal studies. The electrical conductance studies of the complexes in DMF in 10(-3) M concentration indicate their non-electrolytic nature. Room temperature magnetic susceptibility measurements revealed diamagnetic nature of the complexes. Electronic absorption spectra of the complexes show intra-ligand and charge transfer transitions, respectively. Bonding of the metal ion through N- and O- donor atoms of the ligands is revealed by IR studies and the chemical environment of the protons is also confirmed by NMR studies. The thermal analysis data of the complexes indicate the presence of coordinated water molecules. The agar cup and tube dilution methods have been used to study the antibacterial activity of the complexes against the pathogenic bacteria S. aureus, C. diphtherinae, S. typhi and E. coli.
Asunto(s)
Aminoácidos/síntesis química , Aminoácidos/farmacología , Antibacterianos/síntesis química , Antibacterianos/farmacología , Óxidos/síntesis química , Óxidos/farmacología , Oxiquinolina/síntesis química , Oxiquinolina/farmacología , Uranio/farmacología , Bacterias/efectos de los fármacos , Bacterias/crecimiento & desarrollo , Pruebas Antimicrobianas de Difusión por Disco , Conductividad Eléctrica , Ligandos , Espectroscopía de Resonancia Magnética , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Oxiquinolina/análogos & derivados , Espectrofotometría Ultravioleta , Espectroscopía Infrarroja por Transformada de Fourier , Relación Estructura-Actividad , Temperatura , Agua/químicaRESUMEN
If a new generation of iron chelators specifically devoted for cancer chemotherapy emerged these last years, any of them has not yet been approved at this time. Accordingly, there is a need to optimize new chelating molecules for iron chelation therapy and cancer treatment. So, the objective of the present investigation was to characterize the antiproliferative activity and the iron chelating capacity of the iron chelator S1 [bis-N-(8-hydroxyquinoline-5-ylmethyl)benzylamine]. Its effects were compared to O-trensox which binds ferric iron with a very high affinity (pFe(3+)=29.5). For this purpose, primary rat hepatocyte stimulated by EGF and human hepatoma HepaRG cell cultures were used. In these models, the anti-proliferative effect, the inhibition of DNA synthesis and the iron-chelating efficiency of increasing concentrations of S1 and O-trensox (0 up to 200 µM) were investigated. In the two cell culture models, we observed that S1 was about 100 times more efficient than O-trensox and the antiproliferative effect of S1 in HepaRG cells appeared at concentrations as low as 0.1 µM without cytotoxicity. Moreover, the stoichiometry of S1 for iron seemed to be in the range S1/Fe(3+)=1. Using the calcein fluorescence assay, we demonstrated that the affinity of S1 for iron was better than that of O-trensox since it was at least two times more effective to restore the fluorescence of calcein previously quenched by iron. So, the iron chelating efficiency of S1 could explain at least partially its higher anti-proliferative effect compared to O-trensox. Finally, these results suggest that molecules such as S1 may constitute a promising starting point to improve cancer treatment.
Asunto(s)
Bencilaminas/farmacología , Carcinoma Hepatocelular/tratamiento farmacológico , Proliferación Celular/efectos de los fármacos , Hepatocitos/efectos de los fármacos , Quelantes del Hierro/farmacología , Oxiquinolina/farmacología , Animales , Bencilaminas/síntesis química , Bencilaminas/química , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Relación Dosis-Respuesta a Droga , Etilaminas/química , Etilaminas/farmacología , Hepatocitos/metabolismo , Humanos , Hidroxiquinolinas/química , Hidroxiquinolinas/farmacología , Quelantes del Hierro/síntesis química , Quelantes del Hierro/química , Masculino , Oxiquinolina/análogos & derivados , Oxiquinolina/síntesis química , Oxiquinolina/química , Ratas , Ratas Sprague-Dawley , Pruebas de ToxicidadRESUMEN
Metal ion-DNA interactions are important in nature, often changing the genetic material's structure and function. A new Yb complex of YbCl(3) (tris(8-hydroxyquinoline-5-sulfonic acid) ytterbium) was synthesized and utilized as an electrochemical indicator for the detection of DNA oligonucleotide based on its interaction with Yb(QS)(3). Cyclic voltammetry (CV) and fluorescence spectroscopy were used to investigate the interaction of Yb(QS)(3) with ds-DNA. It was revealed that Yb(QS)(3) presented an excellent electrochemical activity on glassy carbon electrode (GCE) and could intercalate into the double helix of double-stranded DNA (ds-DNA). The binding mechanism of interaction was elucidated on glassy carbon electrode dipped in DNA solution and DNA modified carbon paste electrode by using differential pulse voltammetry and cyclic voltammetry. The binding ratio between this complex and ds-DNA was calculated to be 1:1. The extent of hybridization was evaluated on the basis of the difference between signals of Yb(QS)(3) with probe DNA before and after hybridization with complementary DNA. With this approach, this DNA could be quantified over the range from 1 × 10(-8) to 1.1 × 10(-7)M. The interaction mode between Yb(QS)(3) and DNA was found to be mainly intercalative interaction. These results were confirmed with fluorescence experiments.
Asunto(s)
Técnicas Biosensibles/métodos , ADN/análisis , Electroquímica/métodos , Sustancias Intercalantes/metabolismo , Oxiquinolina/análogos & derivados , Iterbio/metabolismo , Carbono/química , ADN/química , Electrodos , Sustancias Intercalantes/química , Oxiquinolina/química , Potenciometría , Espectrometría de Fluorescencia , Iterbio/químicaRESUMEN
Postoperative ileus (POI) is a transient bowel dysmotility that occurs following many types of operations and is one of the most common complications of gastrointestinal surgery. We hypothesized that enteral supplementation of glutamine after abdominal surgery would restore fuel to the small intestine, suppress oxidative stress, and lead to improvement in POI. Twelve dogs underwent distal gastrectomy and were each randomly assigned to one of two groups based on postoperative treatment: the water injection (control) group and the glutamine injection group. Water (40 ml) or L(+)-glutamine (1 g/40 ml water) was injected into the residual stomach through the gastric tube every 12 h after surgery for 7 days. Changes in the plasma and intestinal intracellular concentration of glutamine and in gastrointestinal motility were measured. The plasma and intracellular glutamine levels decreased after the operation in both groups, although the decreased intracellular glutamine levels were not significantly different than preoperative levels. The glutamine group showed a significantly smaller decrease of the plasma glutamine level compared with the control group (P < 0.05). All the dogs showed gastrointestinal dysmotility after the operation. The mean length of time between the operation and the appearance of interdigestive migrating contractions in the glutamine group was significantly shorter than in the control group (22.4 +/- 3.1 h versus 37.8 +/- 4.0 h, respectively; P < 0.05). In conclusion, glutamine could act as a motility-recovery agent after abdominal surgery and thereby decrease the duration of POI.
Asunto(s)
Gastrectomía/efectos adversos , Motilidad Gastrointestinal/efectos de los fármacos , Glutamina/farmacología , Complicaciones Posoperatorias/tratamiento farmacológico , Animales , Perros , Combinación de Medicamentos , Femenino , Glutamina/administración & dosificación , Glutamina/sangre , Oxiquinolina/análogos & derivados , QuinaldinasRESUMEN
Neurodegenerative disorders include a variety of pathological conditions, which share similar critical metabolic processes such as protein aggregation and oxidative stress, both of which are associated with the involvement of metal ions. Chelation therapy could provide a valuable therapeutic approach to such disease states, since metals, particularly iron, are realistic pharmacological targets for the rational design of new therapeutic agents.
Asunto(s)
Quelantes del Hierro/uso terapéutico , Enfermedades Neurodegenerativas/tratamiento farmacológico , Secuencia de Aminoácidos , Péptidos beta-Amiloides/química , Animales , Humanos , Datos de Secuencia Molecular , Oxiquinolina/análogos & derivados , Oxiquinolina/uso terapéutico , Fragmentos de Péptidos/química , Piridinas/uso terapéuticoRESUMEN
Alumina nanotubes were fabricated by a template method. Tris-(8-hydroxyquinoline) gallium (GaQ3) organic nanowires were used as a soft template for coating with alumina using an atomic layer deposition technique. The deposition was conducted at 25 degrees C by using trimethylaluminum and distilled water as the precursors of Al2O3. Amorphous alumina nanotubes were obtained after removing the GaQ3 by dissolving in toluene or by heat treatment at 350 degrees C. The amorphous nanotubes could be crystallized by heating at 900 degrees C for 1 h in vacuum.
Asunto(s)
Óxido de Aluminio/química , Cristalización/métodos , Nanotecnología/métodos , Nanotubos/química , Nanotubos/ultraestructura , Compuestos Organometálicos/química , Oxiquinolina/análogos & derivados , Sustancias Macromoleculares/química , Ensayo de Materiales , Conformación Molecular , Oxiquinolina/química , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
PURPOSE: Cell-based therapy by transplantation of progenitor cells has emerged as a promising development for organ repair, but non-invasive imaging approaches are required to monitor the fate of transplanted cells. Radioactive labelling with (111)In-oxine has been used in preclinical trials. This study aimed to validate (111)In-oxine labelling and subsequent in vivo and ex vivo detection of haematopoietic progenitor cells. METHODS: Murine haematopoietic progenitor cells (10(6), FDCPmix) were labelled with 0.1 MBq (low dose) or 1.0 MBq (high dose) (111)In-oxine and compared with unlabelled controls. Cellular retention of (111)In, viability and proliferation were determined up to 48 h after labelling. Labelled cells were injected into the cavity of the left or right cardiac ventricle in mice. Scintigraphic images were acquired 24 h later. Organ samples were harvested to determine the tissue-specific activity. RESULTS: Labelling efficiency was 75 +/- 14%. Cellular retention of incorporated (111)In after 48 h was 18 +/- 4%. Percentage viability after 48 h was 90 +/- 1% (control), 58 +/- 7% (low dose) and 48 +/- 8% (high dose) (p<0.0001). Numbers of viable cells after 48 h (normalised to 0 h) were 249 +/- 51% (control), 42 +/- 8% (low dose) and 32 +/- 5% (high dose) (p<0.0001). Cells accumulated in the spleen (86.6 +/- 27.0% ID/g), bone marrow (59.1 +/- 16.1% ID/g) and liver (30.3 +/- 9.5% ID/g) after left ventricular injection, whereas most of the cells were detected in the lungs (42.4 +/- 21.8% ID/g) after right ventricular injection. CONCLUSION: Radiolabelling of haematopoietic progenitor cells with (111)In-oxine is feasible, with high labelling efficiency but restricted stability. The integrity of labelled cells is significantly affected, with substantially reduced viability and proliferation and limited migration after systemic transfusion.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/diagnóstico por imagen , Radioisótopos de Indio , Compuestos Organometálicos , Oxiquinolina/análogos & derivados , Radiofármacos , Animales , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Modelos Animales de Enfermedad , Femenino , Ventrículos Cardíacos/efectos de los fármacos , Ventrículos Cardíacos/efectos de la radiación , Células Madre Hematopoyéticas/efectos de la radiación , Pulmón/efectos de los fármacos , Pulmón/efectos de la radiación , Ratones , Ratones Endogámicos C57BL , Cintigrafía/métodos , Factores de TiempoRESUMEN
PURPOSE: To determine the feasibility of in vivo localization and quantification of indium 111 (111In)-oxine-labeled bone marrow (BM) with high-resolution whole-body helical single photon emission computed tomography (SPECT) in an established murine model of atherosclerosis and vascular repair. MATERIALS AND METHODS: The institutional animal care and use committee approved this study. BM from young B6 Rosa 26 Lac Z+/+ mice was radiolabeled with 111In-oxine. On days 1, 4, and 7 after administration of radiolabeled cells, five C57/BL6 apolipoprotein E-deficient mice and five wild-type (WT) control mice were imaged with whole-body high-resolution helical SPECT. Quantification with SPECT was compared with ex vivo analysis by means of gamma counting. Autoradiography and beta-galactosidase staining were used to verify donor cell biodistribution. Linear regression was used to assess the correlation between continuous variables. Two-tailed Student t test was used to compare values between groups, and paired two-tailed t test was used to assess changes within subjects at different time points. RESULTS: SPECT image contrast was high, with clear visualization of BM, liver, and spleen 7 days after administration of radiolabeled cells. SPECT revealed that 42% and 58% more activity was localized to the aorta and BM (P<.05 for both), respectively, in apolipoprotein E-deficient mice versus WT mice. Furthermore, 28% and 27% less activity was localized to the liver and spleen (P<.05 for both), respectively, in apolipoprotein E-deficient mice versus WT mice. SPECT and organ gamma counts showed good quantitative correlation (r=0.9). beta-Galactosidase staining and microautoradiography of recipient aortas showed donor cell localization to the intima of visible atherosclerotic plaque but not to unaffected regions of the vessel wall. CONCLUSION: High-resolution in vivo helical pinhole SPECT can be used to monitor and quantify early biodistribution of 111In-oxine-labeled BM in a murine model of progenitor cell therapy for atherosclerosis.
Asunto(s)
Aterosclerosis/diagnóstico por imagen , Aterosclerosis/cirugía , Células de la Médula Ósea/diagnóstico por imagen , Trasplante de Médula Ósea/diagnóstico por imagen , Aumento de la Imagen/métodos , Compuestos Organometálicos , Oxiquinolina/análogos & derivados , Tomografía Computarizada de Emisión de Fotón Único/métodos , Animales , Trasplante de Médula Ósea/métodos , Modelos Animales de Enfermedad , Estudios de Factibilidad , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , RadiofármacosRESUMEN
The development of tumor vasculature is thought to occur through two complementary processes: sprouting angiogenesis from preexisting blood vessels of the host, and vasculogenesis, which involves the spontaneous development of vessels through specific recruitment, differentiation, and vascular incorporation of circulating endothelial cells (EC), endothelial progenitor cells (EPC), or potentially bone marrow-derived cells. Recent reports, however, have challenged the belief that bone marrow-derived cells contribute to tumor neovascularization, claiming an exclusive role for sprouting angiogenesis in tumor blood vessel development. In the present study, we explored the recruitment behavior of bone marrow-derived lin(-)c-kit(+)Sca-1+ stem cells to subcutaneously implanted Lewis lung carcinoma in a syngeneic bone marrow transplantation model. We observed that although lin(-)c-kit(+)Sca-1+ and their derived cells demonstrate significant recruitment to carcinomas in vivo, they do not appear to functionally contribute to tumor neovascularization. Furthermore, our results support the hypothesis that new vessel formation in carcinomas occurs primarily through endothelialization from adjacent and preexisting vasculature.
Asunto(s)
Antígenos Ly/genética , Antígenos Ly/fisiología , Células de la Médula Ósea/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/fisiología , Neovascularización Patológica , Oxiquinolina/análogos & derivados , Proteínas Proto-Oncogénicas c-kit/genética , Proteínas Proto-Oncogénicas c-kit/fisiología , Animales , Médula Ósea/patología , Células de la Médula Ósea/citología , Trasplante de Médula Ósea , Carcinoma , Carcinoma Pulmonar de Lewis/patología , Diferenciación Celular , Línea Celular Tumoral , Linaje de la Célula , Células Endoteliales/metabolismo , Endotelio Vascular/patología , Citometría de Flujo , Técnica del Anticuerpo Fluorescente Indirecta , Ratones , Ratones Endogámicos C57BL , Microscopía Fluorescente , Trasplante de Neoplasias , Compuestos Organometálicos/química , Oxiquinolina/química , Células Madre/citología , Distribución TisularRESUMEN
BACKGROUND AND OBJECTIVES: In accordance with Food and Drug Administration (FDA) regulations, platelets can be stored in the liquid state at 22 degrees C for only 5 days. Platelets frozen with 6% dimethylsulphoxide (DMSO) can be stored at -80 degrees C for 2 years, and platelets frozen with 5% DMSO can be stored at -150 degrees C for 3 years. Studies are being conducted to determine the effects of lyophilization of platelets. In the present study, we assessed the survival of autologous lyophilized-reconstituted platelets in the baboon. MATERIALS AND METHODS: We studied fresh baboon platelets and baboon platelets that had been treated with paraformaldehyde, frozen, lyophilized, thawed and reconstituted. Aliquots of these platelets were labelled with (111)In-oxine or biotin-X-N-hydroxysuccinimide (biotin-X-NHS) before autotransfusion, and measurements were made of the in vivo recovery and lifespan. We also evaluated the response of fresh and lyophilized platelets to in vitro agonists by measuring the level of platelet surface markers and heterotypic aggregates in the peripheral blood following the autotransfusions. RESULTS: The (111)In-oxine- or biotin-X-NHS-labelled lyophilized, reconstituted platelets exhibited survival times of less than 15 min. These platelets did not respond to stimulation with agonists to decrease platelet GPIb and increase platelet P-selectin and platelet GPIIb-IIIa levels 1 min post-transfusion and they accumulated more procoagulant factor V than did the fresh platelets. CONCLUSIONS: Lyophilized reconstituted baboon platelets labelled with (111)In-oxine or biotin-X-NHS before autotransfusion exhibited an in vivo circulation time of less than 15 min. Further study of the lyophilized, reconstituted platelets is required to evaluate their haemostatic function.
Asunto(s)
Biotina/análogos & derivados , Plaquetas/citología , Transfusión de Sangre Autóloga , Liofilización , Oxiquinolina/análogos & derivados , Transfusión de Plaquetas , Animales , Plaquetas/química , Conservación de la Sangre/métodos , Conservación de la Sangre/normas , Supervivencia Celular , Masculino , Compuestos Organometálicos , Selectina-P/análisis , Papio , Activación Plaquetaria , Complejo GPIb-IX de Glicoproteína Plaquetaria/análisis , SuccinimidasRESUMEN
BACKGROUND AND OBJECTIVES: Studies were carried out in five healthy male baboons to determine the 111indium oxine (111In-oxine) survival of autologous fresh, liquid-preserved and cryopreserved platelets. Simultaneous organ-distribution studies were performed to determine the percentage uptake of platelets by the spleen and/or liver. MATERIALS AND METHODS: Each of five baboons was transfused, on three different occasions, with autologous fresh platelets stored at 22 degrees C for 18 h, liquid-preserved platelets stored at 22 degrees C for 5 days and washed previously frozen platelets, labelled with 111In-oxine. RESULTS: In vivo recovery at 2 h was 81% for the fresh platelets, 54% for the previously frozen platelets and 44% for the 5-day-old liquid-preserved platelets. The weighted mean life span was 5.4 days for fresh platelets, 4.2 days for previously frozen platelets and 2 days for liquid preserved platelets. Increased radioactivity was detected over the liver 2 h after transfusion for both the previously frozen and liquid-preserved platelets. CONCLUSIONS: Cryopreserved platelets and liquid-preserved platelets stored at 22 degrees C for 5 days had reduced survival 2 h post-transfusion and reduced life span values compared to fresh platelets. In addition, the finding of increased radioactivity over the liver in the baboons that received cryopreserved and liquid-preserved platelets suggested that the liver was the site for removal of the non-viable platelets.
Asunto(s)
Circulación Sanguínea , Plaquetas/citología , Conservación de la Sangre , Criopreservación , Oxiquinolina/análogos & derivados , Transfusión de Plaquetas , Animales , Transfusión de Sangre Autóloga , Movimiento Celular , Supervivencia Celular , Senescencia Celular , Hígado , Masculino , Compuestos Organometálicos/farmacocinética , Oxiquinolina/farmacocinética , Papio , Trazadores Radiactivos , BazoRESUMEN
Immune thrombocytopenic purpura (ITP) is an autoimmune disease related to the presence of elevated levels of platelet-associated immunoglobulin, or autoantibodies. In recent years the importance of macrophage Fc gamma receptors in the uptake of platelets in ITP has been confirmed. Although in patients with ITP the platelet destruction occurs in liver and spleen, in this present experimental mouse model the liver was the principal organ of sequestration of sensitized platelets. The uptake in the spleen, bone marrow, lung, and kidneys was negligible and not different from that in control animals. In addition, the trapped platelets did not return to circulation, and new cells derived from the platelet-storage pool or new thrombocytogenesis were necessary to restore the platelet count. The depletion of splenic and hepatic murine macrophages by liposome-encapsulated clodronate (lip-clod) was studied as a new strategy for ITP treatment. Lip-clod inhibits, in a dose-dependent manner, the antibody-induced thrombocytopenia. Moreover, lip-clod treatment rapidly restored (24 hours) the platelet count in thrombocytopenic animals to hematologic safe values, and despite additional antiplatelet antiserum treatment, mice were able to maintain this level of platelets at least up to 48 hours. The bleeding times in lip-clod-treated animals was not different from those in controls, demonstrating that the hemostasis was well controlled in these animals. The results presented in this study demonstrate that lip-clod treatment can be effective in the management of experimental ITP. (Blood. 2000;96:2834-2840)
Asunto(s)
Enfermedades Autoinmunes/tratamiento farmacológico , Ácido Clodrónico/uso terapéutico , Hígado/fisiopatología , Macrófagos/fisiología , Púrpura Trombocitopénica Idiopática/tratamiento farmacológico , Animales , Enfermedades Autoinmunes/fisiopatología , Tiempo de Sangría , Plaquetas/inmunología , Plaquetas/patología , Ácido Clodrónico/administración & dosificación , Ácido Clodrónico/farmacología , Relación Dosis-Respuesta a Droga , Composición de Medicamentos , Evaluación Preclínica de Medicamentos , Recuento de Eritrocitos , Sueros Inmunes/toxicidad , Radioisótopos de Indio , Recuento de Leucocitos , Liposomas , Macrófagos/efectos de los fármacos , Ratones , Modelos Animales , Especificidad de Órganos , Compuestos Organometálicos , Oxiquinolina/análogos & derivados , Recuento de Plaquetas , Púrpura Trombocitopénica Idiopática/inducido químicamente , Púrpura Trombocitopénica Idiopática/fisiopatología , Conejos , Bazo/fisiopatologíaRESUMEN
When infection of implants is suspected, optimal management requires accurate confirmation or exclusion of infection. However, in spite of demonstrative clinical signs cultures of smears or chemical parameters of inflammation frequently are ambiguous. Scintigraphy with indium-labeled white blood cells (WBC) has been reported to be sensitive and specific in the diagnosis of low-grade sepsis of the musculoskeletal system. Twenty-eight patients with possible infection were prospectively studied. Infection was suspected in 19 cases with total hip joint prosthesis, 14 cases with knee joint prosthesis and 1 case with shoulder joint prosthesis. All of them underwent scanning with indium-111-labeled WBC and subsequently underwent surgery. At surgery infections were determined by means of culture or histologic results. When correlated with culture and histologic results sensitivity of 111-indium-WBC imaging was 89% with a specificity of 67% and a predictive accuracy of 77%. In patients with rheumatoid arthritis, however, predictive accuracy of 111-indium-labeled WBC imaging was higher than with standard diagnostic methods. The difference was statistically significant (P < 0.05, chi(2)-test). In the patients examined as a whole, predictive accuracy of 111-indium-labeled WBC imaging does not differ from that of standard diagnostic methods. That is why expensive and time-consuming 111-indium-WBC imaging is not justified generally in diagnosis of infection of implants. 111-Indium-WBC imaging is well suited to supplement standard diagnostic methods in patients with rheumatoid arthritis.
Asunto(s)
Artritis/diagnóstico por imagen , Prótesis de Cadera/efectos adversos , Prótesis Articulares/efectos adversos , Prótesis de la Rodilla/efectos adversos , Leucocitos/diagnóstico por imagen , Infecciones Relacionadas con Prótesis/diagnóstico por imagen , Articulación del Hombro , Adulto , Anciano , Enfermedad Crónica , Reacciones Falso Negativas , Reacciones Falso Positivas , Humanos , Radioisótopos de Indio , Persona de Mediana Edad , Compuestos Organometálicos , Oxiquinolina/análogos & derivados , Cintigrafía , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: The inhibiting effect of nitric oxide on the aggregation and adhesion of neutrophils and platelets has been well documented in vitro. In vivo evidence, however, is more scant. In this study, we studied the effects of inhaled nitric oxide on pulmonary cellular sequestration in our sham hemodialysis model. Accumulation of neutrophils and platelets in the lungs has been shown to be an early event in this model. DESIGN: Prospective, randomized, controlled study. SETTING: Animal laboratory at a university medical center. SUBJECTS: Twenty-six anesthetized, mechanically ventilated pigs. INTERVENTIONS: 111Indium-oxine was used to selectively label neutrophils or platelets and the activity over the lungs was followed dynamically with a gamma camera. Sham hemodialysis, using a cuprophan hollow-fiber dialyzer, was instituted via catheters in the femoral vessels. The animals were divided into two main groups: a) the nitric oxide recipient group (n = 12, with platelets labeled in seven animals and neutrophils labeled in five animals); and b) the control group (n = 14, with platelets labeled in seven animals and neutrophils labeled in seven animals). The animals in the former group were given 50 parts per million of nitric oxide in the inspiratory gas from the beginning of dialysis and for 30 mins onward. MEASUREMENTS AND MAIN RESULTS: Inhalation of nitric oxide attenuated the increase in activity over the lungs in both the neutrophil and platelet groups during sham hemodialysis. In addition, an inhibiting effect on the increase in pulmonary pressure was noted. CONCLUSION: Apart from the effects of nitric oxide on central hemodynamics in this model, the scintigraphic findings indicate an in vivo effect of nitric oxide on the accumulation of platelets and neutrophils in the lungs, probably due to inhibition of the adhesion and/or aggregation of these cells.
Asunto(s)
Neutrófilos/efectos de los fármacos , Óxido Nítrico/farmacología , Adhesividad Plaquetaria/efectos de los fármacos , Agregación Plaquetaria/efectos de los fármacos , Diálisis Renal/efectos adversos , Adhesividad/efectos de los fármacos , Administración por Inhalación , Animales , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Radioisótopos de Indio , Compuestos Organometálicos , Oxiquinolina/análogos & derivados , Inhibidores de Agregación Plaquetaria/farmacología , Distribución Aleatoria , Porcinos , Factores de TiempoRESUMEN
Photodynamic therapy is being investigated as an adjuvant treatment for intracranial neoplasms. The efficacy of this therapy is based on the uptake of photosensitizer by neoplastic tissue, its clearance from surrounding brain tissue, and the timing and placement of photoactivating sources. Photofrin-II is the photosensitizer most actively being investigated. We labeled Photofrin-II with Indium-111 and studied the uptake and distribution of this agent in 20 patients with intracranial neoplasms, using single photon emission computed tomography (SPECT) with volume rendering in three dimensions. Of these patients, 16 had malignant glial tumors, 2 had metastatic deposits, 1 had a chordoma, and 1 had a meningioma. Anatomical-spatial data correlated well between the SPECT images and contrast-enhanced computed tomography or magnetic resonance images. Regions of focal uptake on SPECT images correlated with the surgical histopathological findings of the neoplasm. The kinetics of photosensitizer uptake varied according to the tumor's histological findings, the patient's use of steroids, and among patients with similar types of tumor histology. Peak ratios of target-to-nontarget tissue varied from 24 to 72 hours after injection. The study data show that, to be most effective, photodynamic therapy may need to be tailored for each patient by correlating SPECT images with anatomical data produced by computed tomography or magnetic resonance images. Photoactivating sources then can be placed, using computer-assisted stereotactics, to activate a prescribed volume of photosensitized tumor at the optimal time for treatment.
Asunto(s)
Neoplasias Encefálicas/tratamiento farmacológico , Éter de Dihematoporfirina/farmacocinética , Fotorradiación con Hematoporfirina/instrumentación , Tomografía Computarizada de Emisión de Fotón Único/instrumentación , Adulto , Anciano , Astrocitoma/diagnóstico por imagen , Astrocitoma/tratamiento farmacológico , Encéfalo/diagnóstico por imagen , Encéfalo/efectos de los fármacos , Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/secundario , Éter de Dihematoporfirina/administración & dosificación , Femenino , Glioblastoma/diagnóstico por imagen , Glioblastoma/tratamiento farmacológico , Glioma/diagnóstico por imagen , Glioma/tratamiento farmacológico , Humanos , Neoplasias Pulmonares/diagnóstico por imagen , Neoplasias Pulmonares/tratamiento farmacológico , Masculino , Meningioma/diagnóstico por imagen , Meningioma/tratamiento farmacológico , Tasa de Depuración Metabólica/fisiología , Persona de Mediana Edad , Recurrencia Local de Neoplasia/diagnóstico por imagen , Recurrencia Local de Neoplasia/tratamiento farmacológico , Compuestos Organometálicos , Oxiquinolina/análogos & derivadosRESUMEN
Autologous 111In oxine-labelled platelet scintigraphy was used to detect left ventricular thrombi in 20 patients with anterior and 18 patients with inferior Q wave myocardial infarction within 48-72 h. Left ventricular thrombi were found in 8/20 patients with anterior myocardial infarction and in 1/18 patients with inferior myocardial infarction, giving a total incidence of 24%. Patients with left ventricular thrombi were older (64.3 versus 58.2 years), had higher peak creatinine kinase (CK) levels (4523 versus 2749 IU 1-1), lower ejection fraction (19.5 versus 37.8%, P less than 0.005) and were more likely to have an enlarged left ventricle than those without left ventricular thrombi (87.5 versus 54.5%, P less than 0.001). Left ventricular thrombi were found overlying sites of myocardial infarction in 8 out of 9 patients. Apical left ventricular thrombi were 1.7 times more common than septal left ventricular thrombi. All patients received minidose heparin for prevention of deep venous thrombosis. This technique is complementary to echocardiography and may provide additional information in the difficult cases where the decision about full-dose anticoagulation is in doubt.