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1.
Sci Rep ; 12(1): 3171, 2022 Feb 24.
Article in English | MEDLINE | ID: mdl-35210484

ABSTRACT

The analysis of the background noise in seismic networks has proved to be a powerful tool not only to acquire new insights on the crustal structure, but also to monitor different natural and anthropogenic processes. We show that data acquired during controlled source experiments can also be a valuable tool to monitor such processes, in particular when using high-density deployments. Data from a wide-angle reflection and refraction seismic profile in the central-northwest part of Iberia is used to identify signals related to aircrafts, road traffic, quarry blasts, wind blow, rainfall or thunders. The most prominent observations are those generated by a helicopter and an airplane flying following trajectories subparallel to the profile, which are tracked along 200 km with a spatial resolution of 350 m, hence providing an exceptional dataset. Other highlights are the observation of the Doppler effect on signals generated by moving cars and the high-density recording of acoustic waves generated by thunders. In addition to the intrinsic interest of identifying such signals, this contribution proves that it is worth inspecting the data acquired during seismic experiments beyond the time interval including the arrival of the seismic waves generated by the controlled source.

2.
Acta Ortop Mex ; 36(3): 172-178, 2022.
Article in Spanish | MEDLINE | ID: mdl-36862932

ABSTRACT

Implant loosening, catastrophic failure of the bone-screw interface, material migration, and loss of stability of the fixation component assembly constitute a serious complication in adult spinal surgery. The contribution of biomechanics is based on experimental measurement and simulation of transpedicular spinal fixations. The cortical insertion trajectory showed an increase in the resistance of the screw-bone interface with respect to the pedicle insertion trajectory, both for axial traction forces to the screw and for stress distribution in the vertebra. The double-threaded screws and standard pedicle screws had similar strength. Partially threaded screws with four-thread showed better resistance to fatigue in terms of a higher failure load and number of cycles to fail. Cement or hydroxyapatite augmented screws with also showed a better fatigue resistance in osteoporotic vertebrae. Rigid segment simulations confirmed the presence of higher stresses on the intervertebral discs causing damage to adjacent segments. The posterior body of the vertebra may be subjected to high stresses, in the bone-screw interface, being this bone region more susceptible to failure.


El aflojamiento de los implantes, la falla catastrófica de la interfaz hueso-tornillo, la migración de material y la pérdida de estabilidad de los componentes del ensamble de la fijación constituyen una seria complicación en la cirugía de columna de adultos. La contribución de la biomecánica se basa en la medición experimental y la simulación de las fijaciones transpediculares de columna. La trayectoria de inserción cortical mostró un incremento en la resistencia de la interfase tornillo-hueso respecto a la inserción pedicular tanto para fuerzas de tracción axiales al tornillo como en distribución de esfuerzos en la vértebra. Los tornillos de doble rosca tuvieron una resistencia similar a los pediculares estándar. Los tornillos de rosca parcial de cuatro filetes mostraron mejor resistencia a la fatiga en cuanto a mayor carga de rotura y número de ciclos para la falla. Los tornillos aumentados, ya sea con cemento o hidroxiapatita mejoraron también la resistencia a la fatiga en vértebras osteoporóticas. Las simulaciones de segmentos rígidos confirmaron la presencia de esfuerzos superiores en los discos intervertebrales que provocan el daño de los segmentos adyacentes. La parte posterior de las vértebras puede estar sometida a mayores esfuerzos, fundamentalmente en la superficie de la interfaz hueso-tornillo, por lo que son más susceptibles a la falla en esta región.


Subject(s)
Pedicle Screws , Adult , Humans , Biomechanical Phenomena , Bone Cements , Computer Simulation , Software
3.
Med Intensiva (Engl Ed) ; 45(1): 42-55, 2021.
Article in English, Spanish | MEDLINE | ID: mdl-32646669

ABSTRACT

During the new pandemic caused by SARS-CoV-2, there is short knowledge regarding the management of different disease areas, such as coagulopathy and interpretation of D-dimer levels, its association with disseminated intravascular coagulation (DIC) and controversy about the benefit of anticoagulation. Thus, a systematic review has been performed to define the role of D-dimer in the disease, the prevalence of DIC and the usefulness of anticoagulant treatment in these patients. A literature search was performed to analyze the studies of COVID-19 patients. Four recommendations were drawn based on expert opinion and scientific knowledge, according to the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach. The present review suggests the presence of higher levels of D-dimer in those with worse prognosis, there may be an overdiagnosis of DIC in the course of the disease and there is no evidence on the benefit of starting anticoagulant treatment based only on isolated laboratory data.


Subject(s)
Anticoagulants/therapeutic use , Blood Coagulation Disorders/blood , COVID-19/blood , Disseminated Intravascular Coagulation/blood , Fibrin Fibrinogen Degradation Products/analysis , SARS-CoV-2 , Blood Coagulation Disorders/drug therapy , Blood Coagulation Disorders/mortality , COVID-19/epidemiology , COVID-19/mortality , Critical Illness , Disseminated Intravascular Coagulation/diagnosis , Disseminated Intravascular Coagulation/drug therapy , Disseminated Intravascular Coagulation/epidemiology , Humans , Medical Overuse , Observational Studies as Topic , Pandemics , Prevalence , Prognosis , COVID-19 Drug Treatment
4.
Med Intensiva (Engl Ed) ; 44(9): 525-533, 2020 Dec.
Article in English, Spanish | MEDLINE | ID: mdl-32654921

ABSTRACT

OBJECTIVE: To describe the clinical and respiratory characteristics of a cohort of 43 patients with COVID-19 after an evolutive period of 28 days. DESIGN: A prospective, single-center observational study was carried out. SETTING: Intensive care. PATIENTS: Patients admitted due to COVID-19 and respiratory failure. INTERVENTIONS: None. VARIABLES: Automatic recording was made of demographic variables, severity parameters, laboratory data, assisted ventilation (HFO: high-flow oxygen therapy and IMV: invasive mechanical ventilation), oxygenation (PaO2, PaO2/FiO2) and complications. The patients were divided into three groups: survivors (G1), deceased (G2) and patients remaining under admission (G3). The chi-squared test or Fisher exact test (categorical variables) was used, along with the Mann-Whitney U-test or Wilcoxon test for analyzing the differences between medians. Statistical significance was considered for p<0.05. RESULTS: A total of 43 patients were included (G1=28 [65.1%]; G2=10 [23.3%] and G3=5 [11.6%]), with a mean age of 65 years (range: 52-72), 62% males, APACHE II 18 (15-24), SOFA 6 (4-7). Arterial hypertension (30.2%) and obesity (25.6%) were the most frequent comorbidities. High-flow oxygen therapy was used in 62.7% of the patients, with failure in 85%. In turn, 95% of the patients required IMV and 85% received ventilation in prone decubitus. In the general population, initial PaO2/FiO2 improved after 7 days (165 [125-210] vs.194 [153-285]; p=0.02), in the same way as in G1 (164 [125-197] vs. 207 [160-294]; p=0.07), but not in G2 (163 [95-197] vs. 135 [85-177]). No bacterial coinfection was observed. The incidence of IMV-associated pneumonia was high (13 episodes/1000 days of IMV). CONCLUSIONS: Patients with COVID-19 require early IMV, a high frequency of ventilation in prone decubitus, and have a high incidence of failed HFO. The lack of improvement of PaO2/FiO2 at 7 days could be a prognostic marker. .


Subject(s)
COVID-19/epidemiology , Pandemics , SARS-CoV-2 , Age Distribution , Aged , Anti-Bacterial Agents/therapeutic use , Bacterial Infections/complications , Bacterial Infections/drug therapy , COVID-19/mortality , COVID-19/therapy , Chi-Square Distribution , Contraindications, Procedure , Female , Hospital Mortality , Hospitalization/statistics & numerical data , Humans , Intensive Care Units , Male , Middle Aged , Multimorbidity , Noninvasive Ventilation/adverse effects , Prospective Studies , Respiration, Artificial/methods , Spain/epidemiology , Statistics, Nonparametric , Tertiary Care Centers , COVID-19 Drug Treatment
5.
Ann Rheum Dis ; 74(7): 1399-407, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25274633

ABSTRACT

OBJECTIVES: Little is known about targeting the metabolome in non-cancer conditions. Choline kinase (ChoKα), an essential enzyme for phosphatidylcholine biosynthesis, is required for cell proliferation and has been implicated in cancer invasiveness. Aggressive behaviour of fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA) led us to evaluate whether this metabolic pathway could play a role in RA FLS function and joint damage. METHODS: Choline metabolic profile of FLS cells was determined by (1)H magnetic resonance spectroscopy ((1)HMRS) under conditions of ChoKα inhibition. FLS function was evaluated using the ChoKα inhibitor MN58b (IC50=4.2 µM). For arthritis experiments, mice were injected with K/BxN sera. MN58b (3 mg/kg) was injected daily intraperitoneal beginning on day 0 or day 4 after serum administration. RESULTS: The enzyme is expressed in synovial tissue and in cultured RA FLS. Tumour necrosis factor (TNF) and platelet-derived growth factor (PDGF) stimulation increased ChoKα expression and levels of phosphocholine in FLS measured by Western Blot (WB) and metabolomic studies of choline-containing compounds in cultured RA FLS extracts respectively, suggesting activation of this pathway in RA synovial environment. A ChoKα inhibitor also suppressed the behaviour of cultured FLS, including cell migration and resistance to apoptosis, which might contribute to cartilage destruction in RA. In a passive K/BxN arthritis model, pharmacologic ChoKα inhibition significantly decreased arthritis in pretreatment protocols as well as in established disease. CONCLUSIONS: These data suggest that ChoKα inhibition could be an effective strategy in inflammatory arthritis. It also suggests that targeting the metabolome can be a new treatment strategy in non-cancer conditions.


Subject(s)
Arthritis, Rheumatoid/metabolism , Arthritis, Rheumatoid/prevention & control , Butanes/therapeutic use , Choline Kinase/antagonists & inhibitors , Choline Kinase/metabolism , Enzyme Inhibitors/therapeutic use , Pyridinium Compounds/therapeutic use , Animals , Apoptosis/drug effects , Arthritis, Rheumatoid/pathology , Butanes/pharmacology , Cell Movement/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Choline/metabolism , Disease Models, Animal , Enzyme Inhibitors/pharmacology , Fibroblasts/drug effects , Fibroblasts/metabolism , Fibroblasts/pathology , Mice , Mice, Inbred C57BL , Mice, Transgenic , Mitogen-Activated Protein Kinase Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Pyridinium Compounds/pharmacology , Synovial Membrane/drug effects , Synovial Membrane/metabolism , Synovial Membrane/pathology
6.
Nature ; 515(7526): 253-6, 2014 Nov 13.
Article in English | MEDLINE | ID: mdl-25391963

ABSTRACT

Whereas subduction recycling of oceanic lithosphere is one of the central themes of plate tectonics, the recycling of continental lithosphere appears to be far more complicated and less well understood. Delamination and convective downwelling are two widely recognized processes invoked to explain the removal of lithospheric mantle under or adjacent to orogenic belts. Here we relate oceanic plate subduction to removal of adjacent continental lithosphere in certain plate tectonic settings. We have developed teleseismic body wave images from dense broadband seismic experiments that show higher than expected volumes of anomalously fast mantle associated with the subducted Atlantic slab under northeastern South America and the Alboran slab beneath the Gibraltar arc region; the anomalies are under, and are aligned with, the continental margins at depths greater than 200 kilometres. Rayleigh wave analysis finds that the lithospheric mantle under the continental margins is significantly thinner than expected, and that thin lithosphere extends from the orogens adjacent to the subduction zones inland to the edges of nearby cratonic cores. Taking these data together, here we describe a process that can lead to the loss of continental lithosphere adjacent to a subduction zone. Subducting oceanic plates can viscously entrain and remove the bottom of the continental thermal boundary layer lithosphere from adjacent continental margins. This drives surface tectonics and pre-conditions the margins for further deformation by creating topography along the lithosphere-asthenosphere boundary. This can lead to development of secondary downwellings under the continental interior, probably under both South America and the Gibraltar arc, and to delamination of the entire lithospheric mantle, as around the Gibraltar arc. This process reconciles numerous, sometimes mutually exclusive, geodynamic models proposed to explain the complex oceanic-continental tectonics of these subduction zones.

7.
Rev Esp Anestesiol Reanim ; 57(8): 532-5, 2010 Oct.
Article in Spanish | MEDLINE | ID: mdl-21033460

ABSTRACT

To describe the use of the I-gel supraglottic airway device when placing a bronchial blocker for single-lung ventilation, as applied in a preliminary study to explore the feasibility of adopting the technique with appropriate, selected patients undergoing certain thoracic surgery procedures. We used the technique for single-lung ventilation in 25 patients who required isolation of a lung for a variety of thoracic surgical procedures. A bronchial blocker was placed under direct visualization through a fiberoptic bronchoscope and the I-gel supraglottic device. The I-gel mask allowed us to establish a reliable, safe seal of the airway. For an anesthetist with sufficient experience in managing a flexible fiberoptic bronchoscope, it was possible to carry out all procedures without remarkable anesthetic or surgical events. Single-lung ventilation achieved by introducing a bronchial blocker through the I-gel supraglottic device can facilitate safe, effective management of selected patients who must undergo certain thoracic surgery procedures.


Subject(s)
Laryngeal Masks , Respiration, Artificial/instrumentation , Adult , Bronchoscopy , Equipment Design , Female , Glottis , Humans , Male , Respiration, Artificial/methods
8.
Transplant Proc ; 41(6): 2035-43, 2009.
Article in English | MEDLINE | ID: mdl-19715826

ABSTRACT

AIM: The aims of this project were to analyze the factors that influence quality and safety of tissues for transplantation and to develop the method to ensure standards of quality and safety in relation to tissue banking as demanded by European Directive 2004/23/EC and its technical annexes. It is organized in 4 Working Groups, the objectives of each one being focused in a specific area. STANDARDS: The Guide of Recommendations for Tissue Banking is structured into 4 parts: (1) quality systems that apply to tissue banking and general quality system requirements, (2) regulatory framework in Europe, (3) standards available, and (4) recommendations of the fundamental quality and safety keypoints. REGISTRY: This Working Group handled design of a multinational musculoskeletal tissue registry prototype. TRAINING: This Working Group handled design and validation of a specialized training model structured into online and face-to-face courses. The model was improved with suggestions from students, and 100% certification was obtained. AUDIT: The Guide for Auditing Tissue Establishments provides guidance for auditors, a self-assessment questionnaire, and an audit report form. The effectiveness and sustainability of the outputs were assessed. Both guides are useful for experienced tissue establishments and auditors and also for professionals that are starting in the field. The registry prototype proves it is possible to exchange tissues between establishments throughout Europe. The training model has been effective in educating staff and means having professionals with excellent expertise. Member states could adapt/adopt it. The guides should be updated periodically and perhaps a European organization should take responsibility for this and even create a body of auditors.


Subject(s)
Tissue Banks/standards , Certification/standards , Education, Professional , Europe , Guidelines as Topic , Health Planning Guidelines , Humans , Quality Assurance, Health Care , Registries , Safety , Students
11.
Environ Pollut ; 132(3): 479-88, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15325464

ABSTRACT

A field study was performed to evaluate the effect of exposure to organophosphorus (OP) and carbamate (CB) pesticides on the lizard Gallotia galloti palmae. Butyrylcholinesterase (BChE) activity was measured in the plasma of 420 lizards collected from agricultural and reference areas on the Island of La Palma (Canary Islands, Spain) in two sampling periods. Exposure to cholinesterase-inhibiting pesticides was evaluated by a statistical criterion based on a threshold value (two standard deviations below the mean enzyme activity) calculated for the reference group, and a chemical criterion based on the in vitro reactivation of BChE activity using pyridine-2-aldoxime methochloride (2-PAM) or after water dilution of the sample. Mean (+/- SD) BChE activity for lizards from agricultural areas was significantly lower (Fuencaliente site = 2.00 +/- 0.98 micromol min(-1) ml(-1), Tazacorte site = 2.88 +/- 1.08) than that for lizards from the reference areas (Los Llanos site = 3.06 +/- 1.17 micromol min(-1) ml(-1), Tigalate site = 3.96 +/- 1.62). According to the statistical criterion, the number of lizards with BChE depressed was higher at Fuencaliente (22% of males and 25.4% of females) than that sampled at Tazacorte (7.8% of males and 6.2% of females). According to the chemical criterion, Fuencaliente also yielded a higher number of individuals (112 males and 47 females) with BChE activity inhibited by both OP and CB pesticides. CBs appeared to be the pesticides most responsible for BChE inhibition because most of the samples showed reactivation of BChE activity after water treatment (63.3% from Fuencaliente and 29% from Tazacorte). We concluded that the use of reactivation techniques on plasma BChE activity is a better and more accurate method for assessing field exposure to OP/CB pesticides in this lizard species than making direct comparisons of enzyme activity levels between sampling areas.


Subject(s)
Butyrylcholinesterase/blood , Carbamates/toxicity , Lizards/blood , Organophosphorus Compounds/toxicity , Pesticides/toxicity , Animals , Butyrylcholinesterase/drug effects , Cholinesterase Inhibitors/toxicity , Cholinesterase Reactivators/metabolism , Environmental Exposure/adverse effects , Female , Hematocrit , Male , Pralidoxime Compounds/metabolism , Sex Factors
12.
J Pept Res ; 64(2): 51-64, 2004 Aug.
Article in English | MEDLINE | ID: mdl-15251031

ABSTRACT

By screening a solid-phase combinatorial peptide library, a short peptide ligand, YYWLHH, has been discovered that binds with high affinity and selectivity to staphylococcal enterotoxin B (SEB), but only weakly to other SEs that share sequence and structural homology with SEB. Using column affinity chromatography with an immobilized YYWLHH stationary phase, it was possible to separate SEB quantitatively from Staphylococcus aureus fermentation broth, a complex mixture of proteins, carbohydrates and other biomolecules. The immobilized peptide was also used to purify native SEB from a mixture containing denatured and hydrolyzed SEB, and showed little cross-reactivity with other SEs. To our knowledge this is the first report of a highly specific short peptide ligand for SEB. Such a ligand is a potential candidate to replace antibodies for detection, removal and purification strategies for SEB.


Subject(s)
Enterotoxins/metabolism , Ligands , Peptides/chemistry , Peptides/metabolism , Amino Acid Sequence , Humans , Peptide Library , Protein Binding , Staphylococcus aureus/metabolism , Superantigens/metabolism
13.
An Med Interna ; 21(12): 590-2, 2004 Dec.
Article in Spanish | MEDLINE | ID: mdl-15628954

ABSTRACT

Familial mediterranean fever (FMF) is an hereditary disease transmitted in an autosomal recessive way and characterized by recurrent and brief episodes of fever and pain secondary to serositis. The pain is usually located in abdomen simulating an acute abdomen, and in thorax in the form of pleuritic pain. The most severe complication of the FMF is the development of amyloidosis being the main cause of death. This illness affects an specific ethnic group of the mediterranean area, but the prevalence in our area is low. We present the case of a 30 years old man with recurrent thoracic and abdominal pain, whose final diagnostic was FMF. Insisting on the difficulty that it was recognize this proper illness.


Subject(s)
Abdominal Pain/etiology , Familial Mediterranean Fever/complications , Fever/etiology , Adult , Familial Mediterranean Fever/diagnosis , Humans , Male
15.
J Agric Food Chem ; 49(12): 5765-70, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11743761

ABSTRACT

The present work investigates the mechanism of binding of alpha-lactalbumin to the peptide ligand WHWRKR and its variants HWRKR and acetylated WHWRKR immobilized on a polymethacrylate chromatographic resin. The presence of two temperature-dependent binding mechanisms and one temperature-independent mechanism was demonstrated. Injections of different forms of alpha-lactalbumin (apo-alpha-lactalbumin, D87A mutant alpha-lactalbumin) displayed similar behaviors when compared to native alpha-lactalbumin, while lysozyme showed little or no binding to the WHWRKR and AcWHWRKR resins. An alternative process for isolation of alpha-lactalbumin from WPI was shown, using consecutive injections of WPI with limited elution.


Subject(s)
Lactalbumin/metabolism , Oligopeptides/metabolism , Adsorption , Amino Acid Sequence , Ligands , Mutagenesis, Site-Directed , Protein Binding , Protein Precursors/metabolism , Recombinant Proteins/metabolism , Resins, Plant , Thermodynamics
16.
J Chromatogr A ; 874(1): 21-6, 2000 Mar 31.
Article in English | MEDLINE | ID: mdl-10768496

ABSTRACT

Tryptophan enantiomers have been separated by zwitterion pair chromatography using L-leucine-L-leucine-L-leucine peptide as the zwitterion pairing agent. The peptide ligand is adsorbed onto an octadecylsilane support with excess ligand present in bulk solution. This article examines the roles of the hydrophobic matrix and the mobile phase components on tryptophan enantiomer binding and resolution. Capacity factors and enantioselectivites are given for both hydrophobic and hydrophilic matrices using mobile phases containing Leu-Leu-Leu peptide and/or salt. A decrease in selectivity upon the addition of mobile phase salt suggests that quadrupolar ion-pairing contributes to chiral recognition. Results indicate that binding is significantly reduced and separation is not achieved when Leu-Leu-Leu is coupled onto cross-linked or polymerized hydrophilic resins as well as onto macroporous polystyrene resin. However, resin-immobilized Leu-Leu-Asp-Leu-Leu-Leu, Leu-Leu-Glu-Leu-Leu-Leu, and Leu-Leu-Leu-Glu-Leu-Leu peptides, with ion-pairing sites designed to mimic the Leu-Leu-Leu-saturated C18 support, also do not resolve tryptophan enantiomers. This suggests the Leu-Leu-Leu structure is critical for enantiomer resolution. Because D- and L-tryptophan are separated in the absence of bulk Leu-Leu-Leu, chiral discrimination is believed to occur at the surface of the octadecylsilane support.


Subject(s)
Chromatography, High Pressure Liquid/methods , Tryptophan/isolation & purification , Oligopeptides/chemistry , Stereoisomerism , Tryptophan/chemistry
17.
Rev Esp Cardiol ; 53(4): 590-3, 2000 Apr.
Article in Spanish | MEDLINE | ID: mdl-10758039

ABSTRACT

We report the case of a 43 year-old male with a long history of paroxysmal palpitations aborted by vagal manoeuvres, where atrial fibrillation was the only documented arrhythmia. During the electrophysiological study an AV nodal reentrant tachycardia was demonstrated with rapid degeneration into atrial fibrillation. After slow pathway ablation, nodal tachycardia became non-inducible. No palpitations were reported and no arrhythmic event was recorded after a follow-up of 20 months. In selected cases, the electrophysiological study could reveal potentially curable causes of atrial fibrillation.


Subject(s)
Atrial Fibrillation/etiology , Tachycardia, Atrioventricular Nodal Reentry/complications , Adult , Atrial Fibrillation/physiopathology , Electrocardiography , Humans , Male , Tachycardia, Atrioventricular Nodal Reentry/physiopathology
18.
Bioseparation ; 9(6): 385-92, 2000.
Article in English | MEDLINE | ID: mdl-11518242

ABSTRACT

The isolation of individual proteins from whey would allow production of more consistent and reliable products by the food industry and possibly would also increase their use in the pharmaceutical industry. Alpha-lactalbumin is the second most prevalent protein in bovine milk whey and has many uses including serving as an excellent protein source in infant formulas, power drinks and other beverages that require soluble, nutritional protein. In this study, we describe two methods for production of alpha-lactalbumin from whey protein isolate using bioselective adsorption. The use of a peptide ligand (WHWRKR) attached to a resin allowed production of an alpha-lactalbumin-rich fraction with a purity of 90.6% and a recovery of 47.9%, while also producing other fractions of commercial interest. The combined use of an amino resin followed by the WHWRKR resin produce a highly purified alpha-lactalbumin (100%) with a yield of 35.2%.


Subject(s)
Chromatography, Affinity/methods , Milk Proteins/isolation & purification , Animals , Cattle , Lactalbumin/isolation & purification , Ligands , Oligopeptides , Resins, Synthetic , Whey Proteins
19.
J Microencapsul ; 16(5): 647-64, 1999.
Article in English | MEDLINE | ID: mdl-10499844

ABSTRACT

Boron neutron capture therapy (BNCT) is based on the nuclear reaction that occurs when the stable isotope, Boron-10, is irradiated with low-energy thermal neutrons to yield ionizing Helium and Lithium ions that are highly damaging and usually lethal to cells. The successful treatment of cancer by BNCT requires the selective concentration of Boron-10 within malignant tumours. Liposomes have been used as therapeutic compound delivery vehicles for in vivo application, including several anticancer agents. The ability of the boron-containing compound, o-carboranylpropylamine chloride, to accumulate within unilamellar liposomes in response to a transmembrane pH gradient is evaluated. Characterization of the systems obtained is performed for conventional and polyethylene glycol (PEG)-modified (stealth) liposomes, in terms of lipid and CPA contents, vesicle size and stability in detergent solutions. Results demonstrate that CPA loading and vesicle stability can be controlled by the experimental procedure. The loading of CPA into liposomes with average diameters of 100 nm is estimated at 13000 molecules per vesicle for the most stable systems. CPA toxicity to normal human peripheral blood lymphocytes and to adherent glioblastoma multiforme SK-MG-1 cells in vitro is observed to decrease as a result of the entrapment of CPA in liposomes.


Subject(s)
Boron Compounds/chemistry , Boron Compounds/pharmacokinetics , Boron Compounds/toxicity , Boron Neutron Capture Therapy , Cell Adhesion , Detergents/chemistry , Drug Stability , Drug Storage , Glioblastoma/drug therapy , Glioblastoma/metabolism , Glioblastoma/pathology , Humans , Hydrogen-Ion Concentration , Isotopes , Liposomes , Lymphocytes/drug effects , Phosphatidylcholines/chemistry , Phosphatidylcholines/metabolism , Phosphatidylethanolamines/chemistry , Phosphatidylethanolamines/metabolism , Polyethylene Glycols/chemistry , Polyethylene Glycols/metabolism , Tumor Cells, Cultured
20.
Biotechnol Bioeng ; 63(6): 633-41, 1999 Jun 20.
Article in English | MEDLINE | ID: mdl-10397820

ABSTRACT

Affinity chromatography using immobilized S-protein was used for the screening of affinity peptide ligands from two soluble peptide libraries. Peptide library I consisted of octamers with glycine (G) at both termini of each peptide, i.e. GXXXXXXG. The six center positions were constructed using random sequences of six L-amino acids (Y, N, F, E, V, and L). Peptide library II also consisted of octamers but with glycine and valine (V) at both termini of each peptide (GVZZZZVG). The four variable center positions of peptide library II were random sequences of 18 L-amino acids. Peptides that were retained specifically on the immobilized S-protein column were eluted by 2% acetic acid. The peptides in the acid eluate were further separated using reversed-phase HPLC. Each separated peptide fraction was collected and the peptide sequences deconvoluted by mass spectrometry (MS/MS). The screenings of peptide libraries I and II resulted in 12 and 7 affinity peptides, respectively. Eight out of the twelve peptides from peptide library I contained the clear consensus sequence NFEV. Peptide library II resulted in affinity peptides with the sequences GVNFEVVG, GVNFTVVG and GVFFEL(I)VG. The advantages and limitations of affinity chromatography in peptide library screening are discussed.


Subject(s)
Chromatography, Affinity/methods , Peptide Library , Peptides/chemistry , Amino Acid Sequence , Chromatography, High Pressure Liquid/methods , Mass Spectrometry/methods , Peptides/chemical synthesis , Peptides/metabolism , Spectrometry, Mass, Fast Atom Bombardment
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