RESUMEN

BACKGROUND: Oxaliplatin, 5-fluorouracil (5-FU), and leucovorin (LV) are standard first-line treatments for patients with metastatic colorectal cancer (mCRC). The aim of this multicentre, open-label, phase IIIb study was to assess the addition of oxaliplatin to two different 5-FU regimens. PATIENTS AND METHODS: Patients with previously untreated mCRC were randomised to arm A [two-weekly oxaliplatin 85 mg/m(2) + either continuous intravenous infusion (CIV) of 5-FU without LV or two-weekly bolus and CIV 5-FU + LV (LV5FU2)] or arm B (5-FU CIV or LV5FU2 alone). Irinotecan monotherapy was planned on progression. RESULTS: A total of 725 patients were enrolled. After a fixed follow-up of 2 years for each patient, 2-year survival rates were 27.3% and 24.8% in arms A and B, respectively (hazard ratio 0.93; 95% confidence interval 0.78-1.10). The addition of oxaliplatin significantly improved response rates (54.1 versus 29.8%; P < 0.0001) and median progression-free survival (7.9 versus 5.9 months; P < 0.0001). The most common grade 3-4 toxic effects were neutropenia (arm A, 33%; arm B, 5%), diarrhoea (arm A, 14%; arm B, 8%), and fatigue (arm A, 9%; arm B, 8%). CONCLUSIONS: Despite improved rates of tumour control, these results failed to demonstrate a survival benefit from the addition of oxaliplatin to infused 5-FU and lend further support to the use of sequential monotherapy in some patients with mCRC.

Asunto(s)

Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/patología , Fluorouracilo/administración & dosificación , Compuestos Organoplatinos/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Camptotecina/administración & dosificación , Camptotecina/efectos adversos , Camptotecina/análogos & derivados , Camptotecina/uso terapéutico , Neoplasias Colorrectales/mortalidad , Diarrea/inducido químicamente , Progresión de la Enfermedad , Supervivencia sin Enfermedad , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Fatiga/inducido químicamente , Fluorouracilo/efectos adversos , Fluorouracilo/uso terapéutico , Estudios de Seguimiento , Humanos , Infusiones Intravenosas , Irinotecán , Leucovorina/administración & dosificación , Leucovorina/efectos adversos , Leucovorina/uso terapéutico , Metástasis de la Neoplasia , Neutropenia/inducido químicamente , Compuestos Organoplatinos/efectos adversos , Compuestos Organoplatinos/uso terapéutico , Oxaliplatino , Análisis de Supervivencia , Factores de Tiempo , Resultado del Tratamiento

RESUMEN

BACKGROUND: High-dose therapy (HDT) with stem-cell support is the reference treatment for relapsed lymphoma, but is not appropriate for all patients. Conventional salvage chemotherapies have been used with limited efficacy and significant toxicity. Rituximab, gemcitabine and oxaliplatin are active as single agents in relapsed or refractory lymphoma, and have demonstrated synergistic effects in vitro and in vivo. PATIENTS AND METHODS: Forty-six patients with relapsed or refractory B-cell lymphoma received up to eight cycles of R-GemOx (rituximab 375 mg/m(2) on day 1, gemcitabine 1000 mg/m(2) and oxaliplatin 100 mg/m(2) on day 2). The majority (72%) had diffuse large B-cell lymphoma. RESULTS: After four cycles of R-GemOx, the overall response rate was 83% [50% complete response (CR)/unconfirmed CR (CRu)]. High CR/CRu rates were observed in all histological subtypes. In patients who had previously received rituximab, the CR/CRu rate after eight cycles was 65%. The 2-year event-free and overall survival rates (median follow-up of 28 months) were 43% and 66%, respectively. Among responders, the probability of being disease free for 2 years was 62%. Treatment was generally well tolerated. CONCLUSION: R-GemOx shows promising activity with acceptable toxicity in patients with relapsed/refractory B-cell lymphoma who are not eligible for HDT.

Asunto(s)

Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Linfoma de Células B/tratamiento farmacológico , Recurrencia Local de Neoplasia/tratamiento farmacológico , Adulto , Anciano , Anticuerpos Monoclonales/administración & dosificación , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales de Origen Murino , Desoxicitidina/administración & dosificación , Desoxicitidina/efectos adversos , Desoxicitidina/análogos & derivados , Resistencia a Antineoplásicos , Femenino , Humanos , Estimación de Kaplan-Meier , Linfoma de Células B/mortalidad , Masculino , Persona de Mediana Edad , Compuestos Organoplatinos/administración & dosificación , Compuestos Organoplatinos/efectos adversos , Oxaliplatino , Rituximab , Terapia Recuperativa/métodos , Tasa de Supervivencia , Gemcitabina

RESUMEN

BACKGROUND: This multicentre phase II open-label study evaluated safety and antitumour activity of oxaliplatin in cisplatin or carboplatin (cis/carboplatin) +/- taxane-pretreated advanced ovarian cancer (AOC) patients. PATIENTS AND METHODS: Forty-eight patients received oxaliplatin 130 mg/M2 intravenously every 3 weeks, 94% having a performance status (PS) 0-1. All were pretreated with cis/carboplatin and 21 (44%) with paclitaxel. The median number of involved organs was two, 18 (38%) had liver metastasis, 23 (48%) were platinum-resistant and 14 (29%) were taxane-resistant. Forty-two patients were evaluable for a response, 18 (43%) were platinum-resistant and 11 (26%) were taxane-resistant. RESULTS: A total of 253 cycles was administered (median: 5.5/patient). Median cumulative oxaliplatin dose was 666 mg/m2. National Cancer Institute-Common Toxicity Criteria toxicity analysis showed that seven patients (15%) had grade 3/4 thrombocytopenia, two patients (4%) had grade 3 neutropenia, and one patient had grade 3 anaemia. Eleven patients (23%) experienced grade 3 neurosensory toxicity. Of the 29 patients with peripheral neuropathy at the end of treatment, 55% had recovered or improved 1 month later. Eleven objective responses (two complete) were obtained in the 42 evaluable patients [ORR 26%, 95% confidence interval (CI) 14% to 42%], with 10/24 (42%, 95% CI 22% to 63%) in platinum-sensitive, and 1 of 18 (5.6%, 95% CI 0% to 27%) in platinum-resistant patients. Median response duration was 9.2 months (95% CI 6.6% to 11.8%), and median progression-free and overall survival in all treated patients were 4.3 months (95% CI 3.0% to 5.7%) and 15.0 months (95% CI 11.1% to 18.8%), respectively. CONCLUSION: Oxaliplatin has a good safety profile and is active in cis/carboplatin +/- paclitaxel-pretreated AOC patients.

Asunto(s)

Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Compuestos Organoplatinos/uso terapéutico , Neoplasias Ováricas/tratamiento farmacológico , Adulto , Anciano , Antígeno Ca-125/análisis , Carboplatino/administración & dosificación , Cisplatino/administración & dosificación , Femenino , Humanos , Persona de Mediana Edad , Compuestos Organoplatinos/efectos adversos , Oxaliplatino , Paclitaxel/administración & dosificación

RESUMEN

Mass spectrometry (MS) is one of the most powerful instrumental techniques for protein analysis. The electrospray ionization (ESI) approach is known to be very gentle and at the same time compatible with liquid separation techniques such as HPLC and CE. However, ESI is known to be susceptible to salts and impurities, which often cause a dramatic decrease in sensitivity due to the suppression of the ionization of the product of interest. For this reason, LC-ESI-MS coupling has so far been largely limited to reversed-phase chromatography with its hydro-organic mobile phases. Other chromatographic techniques are typically "linked" to ESI-MS by time consuming, off-line desalting steps. On-line microdialysis has been proposed as a solution to this dilemma. In this paper, we introduce an improved microdialysis system, which enlarges the number of putative applications, thus allowing chromatographic separations of biological compounds to be directly coupled to MS detection with little to no loss in time or chromatographic resolution. Examples include separations by affinity, ion-exchange and size-exclusion chromatography, all of which were connected successfully to the ESI-MS detector via the on-line microdialyzer. We propose that, using this system, any kind of chromatography technique can be coupled to ESI-MS, thus enabling for example application in quality control or process monitoring of many bioproduction and downstream processes.

Asunto(s)

Microdiálisis/instrumentación , Proteínas/química , Animales , Bovinos , Cromatografía Líquida de Alta Presión , Cromatografía por Intercambio Iónico , Hemoglobinas/química , Indicadores y Reactivos , Peso Molecular , Mioglobina/química , Proteínas/aislamiento & purificación , Glycine max/química , Espectrometría de Masa por Ionización de Electrospray , Espectrofotometría Ultravioleta , Proteína Estafilocócica A/química , Proteína Estafilocócica A/aislamiento & purificación , Inhibidores de Tripsina/química

RESUMEN

The completion of the human genome project has opened novel scientific avenues in functional genomics, structural genomics and proteomics. These areas have a common goal: the identification of all the proteins acting and cross-talking in a single cell at a defined moment of its lifecycle. The expansion of these areas in bioscience has been facilitated by the rapid development of high throughput screening (HTS) methods which has, in turn, attracted the business community to make investments in this novel business segment of biotechnology. By using these HTS methods, the hope is that novel targets will be validated much more rapidly speeding up the development of novel drugs. Numerous techniques and tools have emerged over the past decade for the identification of small target-specific molecular ligands that exploit a common feature: the exploration of molecular diversity using combinatorial methods. While chemists developed new methods for rapidly and efficiently synthesising and screening large collections of small molecules, biologists used recombinant DNA techniques for selecting displayed repertoires. To this end, the discovery of new low molecular weight peptides is becoming increasingly important, not only as molecular tools for the understanding of protein-protein interactions but also for the generation of lead compounds.

Asunto(s)

Clonación Molecular/métodos , Genoma , Biblioteca de Péptidos , Técnicas Químicas Combinatorias , Humanos

RESUMEN

Single chain Fv antibody fragments (scFv) binding to purified Helicobacter pylori urease were selected from a nonimmune human antibody repertoire displayed on filamentous phage. After three rounds of screening on solid phase urease, 44 clones were found to bind the enzyme and four distinct scFv were identified by sequencing their heavy and light chain variable region genes (V(H) and V(L)). Two of the selected human scFv (scFv B4 and scFv D9) inhibited the activity of H. pylori urease with inhibitory constants (K(i)) of 7 and 2 microM, respectively. Their affinity (K(d)) for H. pylori urease as determined by surface plasmon resonance ranged from 17 to 42 nM. Both scFv were able to bind to urease present on the surface of living H. pylori organisms as demonstrated by flow cytometry analysis. The binding sites of scFv B4 and D9 were mapped by the use of two random hexapeptide libraries (X6 and CX6C) displayed on filamentous bacteriophage. The selected peptide sequences were shown to inhibit scFv binding to H. pylori urease and thus could be used in a vaccination strategy as epitopes mimicking (mimotopes) the region of urease recognized by these human scFv antibody fragments.

Asunto(s)

Anticuerpos Antibacterianos/inmunología , Proteínas Bacterianas/inmunología , Helicobacter pylori/inmunología , Fragmentos de Inmunoglobulinas/inmunología , Ureasa/inmunología , Secuencia de Aminoácidos , Anticuerpos Antibacterianos/aislamiento & purificación , Anticuerpos Antibacterianos/metabolismo , Anticuerpos Antibacterianos/farmacología , Afinidad de Anticuerpos , Especificidad de Anticuerpos , Reacciones Antígeno-Anticuerpo , Proteínas Bacterianas/antagonistas & inhibidores , Proteínas Bacterianas/metabolismo , Vacunas Bacterianas , Diseño de Fármacos , Inhibidores Enzimáticos/inmunología , Inhibidores Enzimáticos/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática , Epítopos/inmunología , Helicobacter pylori/enzimología , Humanos , Fragmentos de Inmunoglobulinas/aislamiento & purificación , Fragmentos de Inmunoglobulinas/metabolismo , Fragmentos de Inmunoglobulinas/farmacología , Imitación Molecular , Datos de Secuencia Molecular , Biblioteca de Péptidos , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Resonancia por Plasmón de Superficie , Ureasa/antagonistas & inhibidores , Ureasa/metabolismo

RESUMEN

Replacement of the hyperimmune anti-Rhesus (Rh) D immunoglobulin, currently used to prevent haemolytic disease of the newborn, by fully recombinant human anti-RhD antibodies would solve the current logistic problems associated with supply and demand. The combination of phage display repertoire cloning with precise selection procedures enables isolation of specific genes that can then be inserted into mammalian expression systems allowing production of large quantities of recombinant human proteins. With the aim of selecting high-affinity anti-RhD antibodies, two human Fab libraries were constructed from a hyperimmune donor. Use of a new phage panning procedure involving bromelin-treated red blood cells enabled the isolation of two high-affinity Fab-expressing phage clones. LD-6-3 and LD-6-33, specific for RhD. These showed a novel reaction pattern by recognizing the D variants D(III), D(IVa), D(IVb), D(Va), D(VI) types I and II. D(VII), Rh33 and DFR. Full-length immunoglobulin molecules were constructed by cloning the variable regions into expression vectors containing genomic DNA encoding the immunoglobulin constant regions. We describe the first, stable, suspension growth-adapted Chinese hamster ovary (CHO) cell line producing a high affinity recombinant human IgG1 anti-RhD antibody adapted to pilot-scale production. Evaluation of the Fc region of this recombinant antibody by either chemiluminescence or antibody-dependent cell cytotoxicity (ADCC) assays demonstrated macrophage activation and lysis of red blood cells by human lymphocytes. A consistent source of recombinant human anti-RhD immunoglobulin produced by CHO cells is expected to meet the stringent safety and regulatory requirements for prophylactic application.

Asunto(s)

Biotecnología/métodos , Fragmentos Fab de Inmunoglobulinas/genética , Inmunoglobulina G/genética , Globulina Inmune rho(D)/metabolismo , Animales , Bacteriófagos , Secuencia de Bases , Bromelaínas/farmacología , Células CHO , Clonación Molecular , Cricetinae , Eritrocitos , Humanos , Fragmentos Fab de Inmunoglobulinas/aislamiento & purificación , Datos de Secuencia Molecular , Proteínas Recombinantes/metabolismo , Isoinmunización Rh/prevención & control

RESUMEN

Urease is an important virulence factor for Helicobacter pylori and is critical for bacterial colonization of the human gastric mucosa. Specific inhibition of urease activity has been proposed as a possible strategy to fight this bacteria which infects billions of individual throughout the world and can lead to severe pathological conditions in a limited number of cases. We have selected peptides which specifically bind and inhibit H. pylori urease from libraries of random peptides displayed on filamentous phage in the context of pIII coat protein. Screening of a highly diverse 25-mer combinatorial library and two newly constructed random 6-mer peptide libraries on solid phase H. pylori urease holoenzyme allowed the identification of two peptides, 24-mer TFLPQPRCSALLRYLSEDGVIVPS and 6-mer YDFYWW that can bind and inhibit the activity of urease purified from H. pylori. These two peptides were chemically synthesized and their inhibition constants (Ki) were found to be 47 microM for the 24-mer and 30 microM for the 6-mer peptide. Both peptides specifically inhibited the activity of H. pylori urease but not that of Bacillus pasteurii.

Asunto(s)

Proteínas Bacterianas/antagonistas & inhibidores , Bacteriófago M13/genética , Helicobacter pylori/enzimología , Biblioteca de Péptidos , Ureasa/antagonistas & inhibidores , Secuencia de Aminoácidos , Proteínas Bacterianas/aislamiento & purificación , Secuencia de Bases , Inhibidores Enzimáticos/farmacología , Helicobacter pylori/química , Holoenzimas/metabolismo , Cinética , Datos de Secuencia Molecular , Péptidos/genética , Péptidos/farmacología , Unión Proteica , Especificidad por Sustrato , Ureasa/aislamiento & purificación

Asunto(s)

ADN/genética , ADN/metabolismo , Programas Informáticos , Factores de Transcripción/metabolismo , Algoritmos , Secuencia de Bases , Sitios de Unión/genética , Simulación por Computador , Humanos , Técnicas In Vitro , Modelos Biológicos

RESUMEN

A new type of high avidity binding molecule, termed "peptabody" was created by harnessing the effect of multivalent interaction. A short peptide ligand was fused via a semi-rigid hinge region with the coiled-coil assembly domain of the cartilage oligomeric matrix protein, resulting in a pentameric multivalent binding molecule. In the first peptabody (Pab-S) described here, a peptide (S) specific for the mouse B-cell lymphoma BCL1 surface Ig idiotype, was selected from a phage display library. A fusion gene was constructed encoding peptide S, followed by the 24 aa hinge region from camel IgG and a modified 55 aa cartilage oligomeric matrix protein pentamerization domain. The Pab-S fusion protein was expressed in Escherichia coli in a soluble form at high levels and purified in a single step by metal-affinity chromatography. Pab-S specifically bound the BCL1 surface idiotype with an avidity of about 1 nM, which corresponds to a 2 x 10(5)-fold increase compared with the affinity of the synthetic peptide S itself. Biochemical characterization showed that Pab-S is a stable homopentamer of about 85 kDa, with interchain disulfide bonds. Pab-S can be dissociated under denaturing and reducing conditions and reassociated as a pentamer with full-binding activity. This intrinsic feature provides an easy way to combine Pab molecules with two different peptide specificities, thus producing heteropentamers with bispecific and/or chelating properties.

Asunto(s)

Proteínas de la Matriz Extracelular , Glicoproteínas/metabolismo , Péptidos/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Animales , Unión Competitiva , Western Blotting , Cromatografía en Gel , Simulación por Computador , Escherichia coli/genética , Glicoproteínas/química , Ligandos , Linfoma de Células B , Proteínas Matrilinas , Ratones , Ratones Endogámicos BALB C , Modelos Moleculares , Péptidos/química , Péptidos/genética , Plásmidos , Unión Proteica , Conformación Proteica , Receptores de Superficie Celular/metabolismo , Proteínas Recombinantes de Fusión/química , Análisis de Secuencia , Transducción de Señal , Células Tumorales Cultivadas

RESUMEN

This is the first study describing recombinant human antibody fragments directed to the autoantigen proteinase 3 (PR3) from an immune B cell source. Detection of these autoantibodies has proven valid for the diagnosis and monitoring of Wegener's granulomatosis. The described antibody fragment (scFv) was isolated from a phage display library prepared from the IgG-positive splenic lymphocytes of a patient with systemic autoimmunity. The cloning strategy was designed to maintain the diversity of the antibody variable gene repertoire, and sequencing of several variable genes demonstrated that all major heavy and light chain families were represented. We found an over-representation of particular heavy chain variable domains in splenic lymphocytes which differ from the ones frequently found in peripheral blood lymphocytes. It was possible to obtain specific scFv to PR3 after a single round of selection and the binding could be inhibited by the patients' sera. Although the antibody fragments in the splenic repertoire were found to be highly mutated, it was interesting to find that the selected scFv showed only limited somatic mutation. Furthermore, we could demonstrate that the removal of the mutations had no effect on binding specificity.

Asunto(s)

Autoantígenos/inmunología , Bacteriófagos/genética , Fragmentos de Inmunoglobulinas/análisis , Serina Endopeptidasas/inmunología , Secuencia de Aminoácidos , Autoinmunidad/genética , Secuencia de Bases , Biblioteca de Genes , Genes de Inmunoglobulinas , Granulomatosis con Poliangitis/inmunología , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/genética , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Mieloblastina

RESUMEN

It has been suggested that recombination and shuffling between exons has been a key feature in the evolution of proteins. We propose that this strategy could also be used for the artificial evolution of proteins in bacteria. As a first step, we illustrate the use of a self-splicing group I intron with inserted lox-Cre recombination site to assemble a very large combinatorial repertoire (> 10(11) members) of peptides from two different exons. Each exon comprised a repertoire of 10 random amino acids residues; after splicing, the repertoires were joined together through a central five-residue spacer to give a combinatorial repertoire of 25-residue peptides. The repertoire was displayed on filamentous bacteriophage by fusion to the pIII phage coat protein and selected by binding to several proteins, including beta-glucuronidase. One of the peptides selected against beta-glucuronidase was chemically synthesized and shown to inhibit the enzymatic activity (inhibition constant: 17 nM); by further exon shuffling, an improved inhibitor was isolated (inhibition constant: 7 nM). Not only does this approach provide the means for making very large peptide repertoires, but we anticipate that by introducing constraints in the sequences of the peptides and of the linker, it may be possible to evolve small folded peptides and proteins.

Asunto(s)

Bacteriófago P1 , Escherichia coli/genética , Exones , Péptidos/química , Péptidos/farmacología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cartilla de ADN , Ensayo de Inmunoadsorción Enzimática , Evolución Molecular , Glucuronidasa/antagonistas & inhibidores , Glucuronidasa/biosíntesis , Intrones , Modelos Genéticos , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Conformación de Ácido Nucleico , Péptidos/síntesis química , Reacción en Cadena de la Polimerasa , ARN Ribosómico/biosíntesis , ARN Ribosómico/genética , Proteínas Recombinantes de Fusión/biosíntesis , Recombinación Genética , Tetrahymena thermophila/genética

RESUMEN

The monoclonal antibody (MAb SEN31, a mouse IgG1 which recognizes the cluster-5a antigen on small-cell lung cancer (SCLC) cells, was used to prepare a selective and potent blocked ricin immunotoxin. In a series of experiments in vitro and in a SCLC xenograft model in nude mice, the tumor localization potential of the radiolabeled antibody SEN31 and the anti-tumor activity of the immunotoxin SEN31-bR, the non-specific binding activity of which had been greatly reduced by blocking of the galactose binding domains of the B-chain, was determined. Radiolabeling of SEN31 was performed by linking a 67Ga-labeled desferrioxamine moiety to the oligosaccharide side chains of the antibody in order to preserve the specific cell-binding activity. 67Ga-SEN31 bound to the antigenic sites on cells of the SW2 SCLC cell line, with a dissociation constant of 3.5 nM and, when injected i.v., selectively localized at the site of s.c.-growing SW2 tumor xenografts in nude mice, with a tumor-to-blood ratio of 3.5. The immunotoxin SEN31-bR was potently and selectively active against SCLC cell lines both of classic and of variant morphologies. At a concentration of 300 pM the immunotoxin selectively eliminated 4.5 logs of clonogenic tumor cells. In nude mice, SEN31-bR was cleared from the blood with biphasic kinetics following i.v. injection and maintained a stable serum level during continuous i.p. infusion. The growth of s.c. SW2 solid-tumor xenografts was delayed following a single i.v. injection or a continuous i.p. infusion, each at a non-toxic dose.

Asunto(s)

Antígenos de Neoplasias/inmunología , Carcinoma de Células Pequeñas/tratamiento farmacológico , Inmunotoxinas/uso terapéutico , Neoplasias Pulmonares/tratamiento farmacológico , Ricina/uso terapéutico , Animales , Anticuerpos Monoclonales/metabolismo , Anticuerpos Monoclonales/uso terapéutico , Carcinoma de Células Pequeñas/inmunología , Femenino , Humanos , Inmunotoxinas/metabolismo , Neoplasias Pulmonares/inmunología , Ratones , Ratones Endogámicos ICR , Ratones Desnudos , Ricina/farmacocinética , Células Tumorales Cultivadas

RESUMEN

Previous experimental results in nude mice showing that radiolabeling the monoclonal antibody anti-CEA 35 with 67Ga-aminooxyacetyldeferroxamine could give better tumor localization than radioiodination prompted us to initiate the present clinical study. The 67Ga-labeled antibody anti-CEA 35 (185 MBq, 0.7-1.7 mg) was injected preoperatively into 14 patients for colorectal carcinoma imaging. The same antibody labeled with 125I (3.7 MBq, 0.25 mg) was injected simultaneously to compare the 67Ga and 125I dose recoveries in surgical specimens. Twelve of 14 primary tumors gave a positive 67Ga scintigraph. The mean %ID/g recovered in all tumors 3-9 days after injection was significantly higher for 67Ga (0.019%) than for 125I (0.005%) (p < 0.001, paired t test). The tumor-to-normal tissue ratios were generally higher for 67Ga, with the exception of liver. We conclude that 67Ga-aminooxyacetyldeferroxamine improved immunoscintigraphy outside the liver, particularly in the pelvic region. We also show that deferroxamine infusion accelerates the excretion of 67Ga in eight patients and propose that this could lead to further improvement of immunoscintigraphy.

Asunto(s)

Neoplasias Colorrectales/diagnóstico por imagen , Deferoxamina , Radioisótopos de Galio , Galio , Radioinmunodetección , Anciano , Antígeno Carcinoembrionario/inmunología , Deferoxamina/análogos & derivados , Femenino , Humanos , Radioisótopos de Yodo , Masculino , Tomografía Computarizada de Emisión de Fotón Único

RESUMEN

We propose a novel method for the site-specific labeling of antibodies under mild conditions and give as an example the modification of an F(ab')2-like fragment of the chimeric monoclonal antibody B72.3. The F(ab')2-like fragment was produced by the action of the protease lysyl endopeptidase. Reverse proteolysis, catalyzed by the same enzyme, was then used to attach carbohydrazide specifically to the carboxyl termini of the heavy chains of the fragment. Finally, a radiolabeled chelator possessing an aldehyde group was conjugated to the modified fragment through a hydrazone linkage. The resulting site-specifically labeled F(ab')2-like fragment was characterized by gel electrophoresis and by enzymic digestion. It was found to possess immunoreactivity equivalent to that of the unmodified F(ab')2-like fragment as determined by immunofluorescence and ELISA (enzyme-linked immunosorbent assay) techniques. The advantages and disadvantages of this labeling method, which appear to be of quite general applicability, are discussed.

Asunto(s)

Anticuerpos Monoclonales/química , Fragmentos Fab de Inmunoglobulinas/química , Anticuerpos Monoclonales/inmunología , Línea Celular , Quelantes , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Hidrazinas/química , Fragmentos Fab de Inmunoglobulinas/inmunología , Radioisótopos de Hierro , Serina Endopeptidasas/metabolismo

RESUMEN

A two-step approach to the production of well-defined protein conjugates is described. In the first step, a linker group, carbohydrazide, having unique reactivity (a hydrazide group) is attached specifically to the carboxyl terminus by using enzyme-catalyzed reverse proteolysis. Since the hydrazide group exists nowhere else on the protein, specificity is assured in a subsequent chemical reaction (formation of a hydrazone bond) of the modified protein with a molecule (chelator, drug, or polypeptide) carrying an aldehyde or keto group. The product is sufficiently stable at neutral pH, no reduction of the hydrazone bond being necessary for the hydrazones described. Protein modification is thus restricted to the carboxyl terminus and a homogeneous product results. With insulin as a model, conditions are described for producing such well-defined conjugates in good yields. The use of other linker groups besides carbohydrazide, and applications of these techniques to antibody fragments, are discussed.

Asunto(s)

Péptido Hidrolasas/química , Proteínas/química , Química Orgánica , Reactivos de Enlaces Cruzados/química , Dimetilsulfóxido , Hidrazinas/síntesis química , Hidrazinas/química , Hidrazonas/síntesis química , Insulina/análogos & derivados , Insulina/química , Fenómenos Químicos Orgánicos , Proteínas/síntesis química , Soluciones , Agua

RESUMEN

In a study of 63 female patients with cervical carcinoma and 37 with corpus carcinoma (all patients from Hamburg), polymorphisms of the blood were determined, which have not previously or only rarely been studied in such patients. Comparisons with a control group revealed no significant differences for the MNSs, P, Jk, SEP, PGM, GPT, EsD, Hp, Gc, and Gm systems. The patients with corpus carcinoma, however, exhibited a lesser frequency of Ccee and an increased frequency of homozygotes in the Fy system. Furthermore, the proportion of patients with GLO1 was greater among the women with cervix carcinoma.

Asunto(s)

Antígenos de Grupos Sanguíneos/genética , Carcinoma/sangre , Polimorfismo Genético , Neoplasias Uterinas/sangre , Antígenos/análisis , Antígenos de Grupos Sanguíneos/inmunología , Eritrocitos/inmunología , Femenino , Humanos , Fenotipo , Neoplasias del Cuello Uterino/sangre

RESUMEN

Both cross-sectional and longitudinal analysis of data from 13,358 women showed that oral contraceptive use is associated with a slight but statistically significant (P lesser than .05) rise in mean blood pressure, which is reversible. The age-adjusted proportion of oral contraceptive users with a blood pressure over 140/90 mm Hg was about three times that on nonusers. These findings are caused by a uniform upward shift in the blood pressure distribution of oral contraceptive users compared to nonusers. Women continuing oral contraceptive use had no appreciably greater change in blood pressure between two visits than persistent nonusers. The clinical implications of a mild contraceptive-induced blood pressure elevation (systolic, 5 to 6 mm Hg; diastolic, 1 to 2 mm Hg) remain unsettled but disturbing.

Asunto(s)

Presión Sanguínea/efectos de los fármacos , Anticonceptivos Orales/efectos adversos , Hipertensión/inducido químicamente , Adolescente , Adulto , Factores de Edad , Peso Corporal , Enfermedades Cardiovasculares/inducido químicamente , Trastornos Cerebrovasculares/inducido químicamente , Complicaciones de la Diabetes , Femenino , Humanos , Hiperlipidemias/complicaciones , Persona de Mediana Edad , Obesidad/complicaciones , Riesgo , Fumar/complicaciones

RESUMEN

Data from 14,961 healthy women were analyzed to determine the relative importance of factors that alter the white blood cell (WBC) count. The effects of smoking, oral contraceptive use, and obesity were most striking. A total leukocyte count greater than 10,00/cu mm was found in 44% of obese, heavily smoking women who took oral contraceptives (central 95% of the distribution was 5,800 to 14,200/cu mm) as compared to 2% of women without these attributes (3,500 to 9,400/cu mm). Other factors such as age, time of day, phase of the menstrual cycle, and red blood cell variables were of lesser significance. The use of noncontraceptive estrogenic hormones did not affect the WBC count. Recognition of these findings is important because excessive laboratory studies, lost physician time, and patient inconvenience may thereby be avoided.

Asunto(s)

Anticonceptivos Orales/farmacología , Obesidad/sangre , Fumar , Adolescente , Adulto , Factores de Edad , Ritmo Circadiano , Femenino , Humanos , Recuento de Leucocitos , Menstruación , Persona de Mediana Edad , Análisis de Regresión

RESUMEN

The relation between oral contraceptive (OC) use, ABO blood groups, and in vitro fibrin formation, as measured by thromboelastography, was investigated in 4315 women. The thromboelastogram (TEG) measures the speed of fibrin formation (R, K) and clot firmness (Ma). More rapid fibrin formation and increased clot firmness were associated with current OC use, but not to the extent seen in pathologic states. This relation was not related to dose, estrogen-progestin type, or duration of exposure. Past and "never" OC users had similar TEG values, suggesting reversibility, and noncontraceptive estrogens had a minimal effect on the TEG. ABO blood groups were found to have only a slight association with TEG values. Subjects with blood group O showed a somewhat prolonged speed of fibrin formation as compared to all other subjects. A decreased clot firmness was found in AB subjects, but this did not persist among OC users.

Asunto(s)

Sistema del Grupo Sanguíneo ABO , Coagulación Sanguínea/efectos de los fármacos , Anticonceptivos Orales/farmacología , Fibrina , Adolescente , Adulto , Factores de Edad , Estrógenos/farmacología , Femenino , Fibrina/fisiología , Humanos , Persona de Mediana Edad , Tromboelastografía , Factores de Tiempo