RESUMEN
Objective: To investigate the levels of sex hormone and fertility in female patients after hematopoietic stem cell transplantation (HSCT), as well as their correlation with conditioning regimens, and analyse the effect of hormone replacement therapy (HRT) in young women after HSCT. Methods: Retrospective case series study. The clinical data of 147 women who underwent HSCT in the First Affiliated Hospital of Soochow University from January 2010 to January 2021 were retrospectively analyzed. The sex hormone levels were measured and followed-up, and the survival, menstrual fertility and the use of HRT of the patients were also followed-up. The sex hormone levels were measured after transplantation, and the ovarian function was evaluated. Independent sample t test and χ2 test were used for comparison between the two groups. Results: The median age of the 147 patients was 26 (range, 10-45) years. Of them, 135 patients received allogeneic HSCT and 12 patients received autologous HSCT. Furthermore, 129 patients received myeloablative conditioning, and 18 patients received reduced conditioning dose. The median follow-up time was 50 months (range, 18-134 months). Five patients died of disease recurrence during follow-up. Of the 54 patients with subcutaneous injection of zoladex, three recovered menstruation spontaneously after transplantation, and all of them were myeloablative conditioning patients, one patient gave birth to twins through assisted reproductive technology. Ninety-three patients did not use zoladex before conditioning, two patients with aplastic anemia with non-myeloablative transplantation resumed menstruation spontaneously, and conceived naturally. The level of follicle stimulating hormone after transplantation in patients receiving myeloablative conditioning regimen was significantly higher than that in patients receiving reduced-dose conditioning regimen [(95.28±3.94) U/L vs. (71.85±10.72) U/L, P=0.039]. Among 147 patients, 122 patients developed premature ovarian failure, 83 patients received sex hormone replacement therapy after transplantation, and 76 patients recovered menstruation and improved endocrine function. Conclusions: The incidence of premature ovarian failure is high in female patients after HSCT, and patients have a chance at natural conception. Reducing the dose of conditioning regimen and the application of zoladex before transplantation can reduce ovarian of conditioning drugs. HRT after transplantation can partially improve the endocrine function of patients.
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Enfermedad Injerto contra Huésped , Trasplante de Células Madre Hematopoyéticas , Insuficiencia Ovárica Primaria , Humanos , Femenino , Niño , Adolescente , Adulto Joven , Adulto , Persona de Mediana Edad , Estudios Retrospectivos , Insuficiencia Ovárica Primaria/etiología , Estudios de Seguimiento , Goserelina , Pronóstico , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Hormonas Esteroides Gonadales , Acondicionamiento Pretrasplante/efectos adversos , Enfermedad Injerto contra Huésped/etiologíaRESUMEN
In the field of asset allocation, how to balance the returns of an investment portfolio and its fluctuations is the core issue. Capital asset pricing model, arbitrage pricing theory, and Fama-French three-factor model were used to quantify the price of individual stocks and portfolios. Based on the second-order stochastic dominance rule, the higher moments of return series, the Shannon entropy, and some other actual investment constraints, we construct a multiconstraint portfolio optimization model, aiming at comprehensively weighting the returns and risk of portfolios rather than blindly maximizing its returns. Furthermore, the whale optimization algorithm based on FTSE100 index data is used to optimize the above multiconstraint portfolio optimization model, which significantly improves the rate of return of the simple diversified buy-and-hold strategy or the FTSE100 index. Furthermore, extensive experiments validate the superiority of the whale optimization algorithm over the other four swarm intelligence optimization algorithms (gray wolf optimizer, fruit fly optimization algorithm, particle swarm optimization, and firefly algorithm) through various indicators of the results, especially under harsh constraints.
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Algoritmos , Ballenas , Animales , Entropía , Inteligencia , Inversiones en SaludRESUMEN
Under the analysis of particle growth mechanism, the monodisperse colloidal silica abrasives for chemical mechanical planarization (CMP) slurry were synthesized by the modified ion-exchanged and hydrothermal step-polymerization process. After the colloidal silica with controllable size was synthesized, its microstructure, stability and CMP performance was characterized and tested by SEM, HRTEM, Zeta potential Analyzer and CMP tester. Results show that the spherical, high stable (Zeta potential: -52.8 mV) colloidal silica with controllable size was achieved. About its CMP performance, the polishing rate for silicon double-side CMP is increased to be 317 nm/min and the polished surface roughness (RMS) was reduced to 0.32 nm.
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Coristoma , Enfermedades del Oído , Trompa Auditiva , Coristoma/cirugía , Enfermedades del Oído/cirugía , Cara , HumanosRESUMEN
AIM: To study the effect of ligustrum fruit on spermatogenesis and blood gonadal hormones in diabetic rats. METHODS: Experimental diabetes was induced in male Wistar rats with streptozotocin. Ligustrum fruit extract was given by gastric gavage at a dose of crude drug 30 g x kg(-1) x d(-1) for 110 days. The serum gonadadotropic hormones and testosterone were determined on d 60 and testicular histology examined on d 110. RESULTS: In the control diabetic rats, the seminiferous tubules were dilated and the spermatogenic cells irregularly arranged. Spermatogenesis was arrested with the number of spermatids highly reduced and spermatozoa not observed. In the treated rats, all types of spermatogenic cells were practically normal. The serum luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone levels were higher in the treated than in the control rats, but the difference was insignificant. CONCLUSION: In experimental diabetic rats, ligustrum fruit extract protects the damaging effect of experimental diabetes on spermatogenesis.
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Diabetes Mellitus Experimental/fisiopatología , Hormonas Esteroides Gonadales/sangre , Extractos Vegetales/farmacología , Espermatogénesis/efectos de los fármacos , Animales , Hormona Folículo Estimulante/sangre , Hormona Luteinizante/sangre , Masculino , Medicina Tradicional China , Ratas , Ratas Wistar , Testosterona/sangreRESUMEN
In order to provide the equivalent of a human anti-human protein antibody as positive control in ELISAs, a goat-human antibody complex was created using chemical cross-linking. The resulting hybrid complex had a larger molecular size on HPLC and SDS-PAGE. In ELISA, the goat-human complex bound to human antigen and was detectable by a secondary anti-human conjugate. The method to make the hybrid complex is simple, cost-effective and can be used to make human-like antibodies to many human proteins.
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Anticuerpos/química , Animales , Cromatografía Líquida de Alta Presión , Reactivos de Enlaces Cruzados , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoglobulinas/análisis , Albúmina Sérica/inmunologíaRESUMEN
The relationships between chronic liver diseases and trace element (TE) contents are debated. Particularly, no defined data are available about the TE levels in viral liver disease patients with or without malnutrition. In this study we evaluated blood and plasma levels of various trace elements in patients with HCV-related chronic liver disease, at different stages of liver damage (8 patients with chronic hepatitis and 32 with liver cirrhosis) with or without malnutrition. We also studied 10 healthy volunteers as control group. We found that cirrhotic subjects had a significant decrease of blood levels of Zn and Se, independently on the nutritional status, whereas plasma levels of Fe were significantly reduced only in malnourished cirrhotic patients. Our data indicate that liver impairment is the main cause of the blood decrease of Se and Zn levels in patients with non alcoholic liver disease, whereas the malnutrition affects Fe levels only.
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Hepatopatías/sangre , Oligoelementos/sangre , Adolescente , Adulto , Anciano , Cromo/sangre , Enfermedad Crónica , Cobre/sangre , Femenino , Humanos , Hierro/sangre , Cirrosis Hepática/sangre , Hepatopatías/complicaciones , Masculino , Manganeso/sangre , Persona de Mediana Edad , Trastornos Nutricionales/sangre , Trastornos Nutricionales/complicaciones , Rubidio/sangre , Selenio/sangre , Zinc/sangreRESUMEN
We evaluated the effect of one year of supplementation with iron plus zinc (12 mg/day of Fe+++ and 12.5 mg/day of Zn++), zinc alone (12.5 mg/day of Zn++) and placebo on growth and on the iron, zinc, copper and selenium tissue contents in 30 well-selected children of short stature (16 M and 14 F; 4-11 years old). Before and after supplementation, we measured the concentrations of iron, transferrin, ferritin, zinc and copper in serum, of zinc in erythrocytes and leukocytes, and of zinc, copper and selenium in hair, as well as glutathione peroxidase activity in erythrocytes. Before supplementation, ferritin and serum, erythrocyte and hair zinc contents were significantly lower than in age-matched controls, while the other measured indices were in the normal range. Iron plus zinc supplementation caused an improvement in growth rate in all subjects, i.e., the median Z-score increased from -2.22 +/- 0.45 to -0.64 +/- 0.55; (p < 0.01). In the zinc-supplemented group, only the subjects whose ferritin levels were higher than 20 ng/L before supplementation showed a similar improvement of growth rate. Iron plus zinc supplementation could be a reasonable treatment in short, prepubertal children affected by marginal zinc and iron deficiency.
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Estatura , Suplementos Dietéticos , Hierro/administración & dosificación , Oligoelementos/análisis , Zinc/administración & dosificación , Determinación de la Edad por el Esqueleto , Antropometría , Niño , Preescolar , Cobre/análisis , Cobre/sangre , Eritrocitos/química , Femenino , Ferritinas/sangre , Glutatión Peroxidasa/sangre , Cabello/química , Humanos , Masculino , Selenio/análisis , Zinc/análisis , Zinc/sangreRESUMEN
Trace elements are involved in chronic liver diseases because these elements may have a direct hepatic toxicity or may be decreased as a consequence of the impaired liver function, particularly in patients with alcoholic cirrhosis and/or malnutrition. In this study, we determined plasma and erythrocytes trace elements in 50 inpatients with nonalcoholic chronic liver disease (11 with biopsy-proven chronic hepatitis, 39 with cirrhosis [16 in stage A according to Child-Pugh criteria, 23 Child B+C]), and in a control group of 10 healthy subjects by the proton induced x-ray emission method. The relationship between trace element concentration and the extent of liver damage, the nutritional status (by anthropometric evaluations), and various blood markers of oxidative stress--reduced glutathione, total lipoperoxides and malonyldialdehyde--was investigated. We found that cirrhotics had a significant decrease of Fe, Zn, Se, and GSH levels in the plasma and of GSH and Se in the erythrocytes with respect to the control and chronic hepatitis groups. GSH levels were related to the degree of liver damage; a significant direct correlation was observed among Se, Zn, and GSH plasma values and between GSH and Se in the erythrocytes. The trace element decrease was, on the contrary, independent of the degree of liver function impairment and only partially affected by the nutritional status. Data indicate that liver cirrhosis, even if not alcohol related, induces a decrease of Se and Zn and that, in these patients, an oxidative stress is present, as documented by the significant correlation between Se and GSH. The plasma Br level was higher in cirrhotics with respect to the control and chronic hepatitis groups.
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Cirrosis Hepática/sangre , Hepatopatías/sangre , Hígado/lesiones , Estrés Oxidativo , Oligoelementos/sangre , Adolescente , Adulto , Anciano , Eritrocitos/metabolismo , Glutatión/sangre , Glutatión/metabolismo , Humanos , Hierro/sangre , Peróxidos Lipídicos/sangre , Malondialdehído/sangre , Persona de Mediana Edad , Fenómenos Fisiológicos de la Nutrición , Selenio/sangre , Rayos X , Zinc/sangreRESUMEN
A new ferulic acid ester, C16H22O9, mp 141-143 degrees C (EtOAc), named sibirate (I), was isolated from the aerial part of Sibiraea angustata (Rchd.) Hand.-Mazz. in addition to a known compound ferulic acid. By means of IR, EI-MS, FAB-MS, 1H NMR, 13C NMR and chemical evidences the structure of sibirate was established as glucityl ferulate.
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Cinamatos/aislamiento & purificación , Medicamentos Herbarios Chinos/química , Sorbitol/análogos & derivados , Cinamatos/química , Estructura Molecular , Sorbitol/química , Sorbitol/aislamiento & purificaciónRESUMEN
Three compounds were isolated from the flower of Lilium devidii. On the basis of chemical reaction, UV, MS, 1HNMR and 13CNMR spectral data, they were identified as beta-sitosterol, stigmasterol and emodin. These compounds are obtained from the plant for the first time.
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Medicamentos Herbarios Chinos/química , Emodina/aislamiento & purificación , Sitoesteroles/aislamiento & purificación , Estigmasterol/aislamiento & purificaciónRESUMEN
As one of the most effective methods to modify proteins, enzymatic hydrolysis is used widely in the preparation of wheat products in the food industry. During the same process, starch pasting occurs frequently. The effects of wheat protein hydrolysis with papain, pepsin, and trypsin on the pasting properties of 3 different kinds of flour were investigated in 5 concentrations. Results showed that the peak viscosity, trough, final, and integral area of pasting curve of these flours decreased with increasing enzymatic hydrolysis of protein, and decreased significantly with the increasing enzyme concentrations. Medium-gluten flour was the least sensitive to enzymatic activity and weak-gluten the most sensitive. Downtrends appeared with increasing papain and trypsin concentrations in the form of breakdown. Enzymes had no significant different effect on the peak times of strong- and medium-gluten flour, but prolonged peak time slightly in weak-gluten flour. The pasting time and temperature of strong- and medium-gluten flour were significantly increased in a concentration-dependent manner. However, there were no significant effects on the pasting times of weak-gluten flour. These results could supply a basis for utilization of enzymatic hydrolysis of wheat protein in food industry and for further studies into the interactions between hydrolyzed protein and starch in food or processing industries.
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Harina/análisis , Glútenes/análisis , Glútenes/metabolismo , Triticum/química , Triticum/enzimología , Manipulación de Alimentos/métodos , Hidrólisis , Papaína/metabolismo , Pepsina A/metabolismo , Almidón/metabolismo , Temperatura , Tripsina/metabolismo , ViscosidadRESUMEN
Polypeptide growth factors promote cellular proliferation by binding to specific plasma membrane-anchored receptors. This interaction triggers the phosphorylation of signal transducing molecules and the transcriptional activation of numerous genes. We have used a differential display approach to identify fibroblast growth factor (FGF)-1-inducible genes in murine NIH 3T3 fibroblasts. Here we report that one of these genes encodes ank, a type IIIa transmembrane protein reported to function in cells as an inorganic pyrophosphate transporter. FGF-1 induction of ank mRNA expression is first detectable at 2 h after growth factor addition and is dependent on de novo RNA and protein synthesis. Ank gene expression is also upregulated after treating quiescent fibroblasts with several other mitogenic agents (e.g., calf serum or platelet-derived growth factor-BB) or the tumor promoter phorbol 12-myristate 13-acetate. Furthermore, in comparison to parental NIH 3T3 cells, oncogene-transformed NIH 3T3 cells constitutively express elevated levels of ank mRNA. FGF-1 also increases ank gene expression in non-immortalized human embryonic lung fibroblasts. Finally, the murine and human ank genes are expressed in vivo in a tissue-specific manner, with highest levels of mRNA expression found in brain, heart, and skeletal muscle. These results indicate that ank is a growth factor-regulated delayed-early response gene in mammalian cells, and we propose that increased ank expression during cell cycle progression may be necessary to maintain proper intracellular pyrophosphate levels during conditions of high cellular metabolic activity.
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Factor 1 de Crecimiento de Fibroblastos/farmacología , Fibroblastos/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Proteínas de la Membrana/genética , Ésteres del Forbol/farmacología , Células 3T3 , Animales , Carcinógenos/farmacología , Línea Celular Transformada , ADN Complementario/análisis , ADN Complementario/aislamiento & purificación , Embrión de Mamíferos/metabolismo , Fibroblastos/metabolismo , Perfilación de la Expresión Génica/métodos , Humanos , Proteínas de la Membrana/metabolismo , Ratones , Proteínas de Transporte de Fosfato , ARN Mensajero/efectos de los fármacos , ARN Mensajero/metabolismo , Distribución TisularRESUMEN
Single-chain Fv proteins are known to aggregate and form multimeric species. We report here that these molecules represent a new class of molecular assembly, which we have termed multivalent Fvs. Each binding site in a multivalent Fv comprises the variable light-chain (VL) domain from a single-chain Fv, and the variable heavy-chain (VH) domain from a second single-chain Fv. Each single-chain Fv in a multivalent Fv is part of two binding sites. We have characterized the multivalent forms of the 4-4-20, CC49 and B6.2 sFvs. The degree of multivalent Fv formation is linker-dependent. Multivalent Fvs cannot form in the absence of an intact linker. Multivalent Fvs can be stabilized by their antigen. The conversion between different forms of the multivalent Fvs can be catalyzed by disassociating agents such as 0.5 M guanidine hydrochloride with 20% ethanol. Multivalent Fvs have significantly different stabilities depending on the specific variable domains from which they are constructed. Two models have been proposed for the structure of a multivalent Fv. We have tested each model by attempting to produce a heterodimer from the anti-fluorescein 4-4-20 and anti-tumor CC49 variable regions. We successfully produced a 4-4-20/CC49 heterodimer that comprises two mixed sFvs. The first mixed sFv is composed of the 4-4-20 VL domain, a 12 residue linker and the CC49 Vh domain. The second mixed sFv is composed of a CC49 VL domain, a 12 residue linker and the 4-4-20 VH domain. The 4-4-20/CC49 heterodimer bound both fluorescein and the tumor-associated glycoprotein-72 antigen. These results support a VH/VL 'rearrangement' model in which each variable domain of a multivalent Fv binding site comes from a different polypeptide chain.
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Especificidad de Anticuerpos , Fragmentos de Inmunoglobulinas/inmunología , Cadenas Pesadas de Inmunoglobulina/inmunología , Cadenas Ligeras de Inmunoglobulina/inmunología , Región Variable de Inmunoglobulina/inmunología , Animales , Antígenos de Neoplasias/metabolismo , Carcinoma , Neoplasias del Colon , Escherichia coli/genética , Femenino , Fluoresceína , Fluoresceínas/metabolismo , Glicoproteínas/metabolismo , Inmunoensayo , Fragmentos de Inmunoglobulinas/genética , Cadenas Pesadas de Inmunoglobulina/genética , Cadenas Ligeras de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/genética , Ratones , Ratones Desnudos , Modelos Inmunológicos , Neoplasias Experimentales , Unión Proteica , Ingeniería de Proteínas , Proteínas Recombinantes/inmunología , SolubilidadRESUMEN
Quantum computing is a quickly growing research field. This article introduces the basic concepts of quantum computing, recent developments in quantum searching, and decoherence in a possible quantum dot realization.
RESUMEN
Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a vascular smooth muscle cell (SMC) mitogen and chemotactic factor that is expressed by endothelial cells, SMCs, monocytes/macrophages, and T lymphocytes. Both the membrane-anchored HB-EGF precursor and the secreted mature HB-EGF protein are biologically active; thus, HB-EGF may stimulate SMC growth via autocrine, paracrine, and juxtacrine mechanisms. In the present study, we report that HB-EGF treatment of serum-starved at aortic SMCs can induce fibroblast growth factor (FGF)-2 (basic FGF) gene expression but not FGF-1 (acidic FGF) gene expression. Increased FGF-2 mRNA expression is first detectable at 1 hour after HB-EGF addition, and maximal FGF-2 mRNA levels, corresponding to an approximately 46-fold level of induction, are present at 4 hours. The effect of HB-EGF on FGF-2 mRNA levels appears to be mediated primarily by a transcriptional mechanism and requires de novo synthesized proteins. HB-EGF induction of FGF-2 mRNA levels can be inhibited by treating cells with the anti-inflammatory glucocorticoid dexamethasone or the glycosaminoglycan heparin. Finally, Western blot analyses indicate that HB-EGF-treated SMCs also produce an increased amount of FGF-2 protein. These results indicate that HB-EGF expressed at sites of vascular injury or inflammation in vivo may upregulate FGF-2 production by SMCs.
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Aorta/metabolismo , Factor de Crecimiento Epidérmico/farmacología , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Músculo Liso Vascular/metabolismo , Animales , Aorta/citología , Northern Blotting , Western Blotting , Células Cultivadas , Cicloheximida/farmacología , Dactinomicina/farmacología , Dexametasona/farmacología , Factor 2 de Crecimiento de Fibroblastos/genética , Heparina/farmacología , Factor de Crecimiento Similar a EGF de Unión a Heparina , Péptidos y Proteínas de Señalización Intercelular , Cinética , Músculo Liso Vascular/citología , Concentración Osmolar , Inhibidores de la Síntesis de la Proteína/farmacología , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas WKY , Transcripción Genética/efectos de los fármacosRESUMEN
Polypeptide growth factors stimulate mammalian cell proliferation by binding to specific cell surface receptors. This interaction triggers numerous biochemical responses including the activation of protein phosphorylation cascades and the enhanced expression of specific genes. We have identified several fibroblast growth factor (FGF)-inducible genes in murine NIH 3T3 cells and recently reported that one of them, the FGF-inducible 14 (Fn14) immediate-early response gene, is predicted to encode a novel, cell surface-localized type Ia transmembrane protein. Here, we report that the human Fn14 homolog is located on chromosome 16p13.3 and encodes a 129-amino acid protein with approximately 82% sequence identity to the murine protein. The human Fn14 gene, like the murine Fn14 gene, is expressed at elevated levels after FGF, calf serum or phorbol ester treatment of fibroblasts in vitro and is expressed at relatively high levels in heart and kidney in vivo. We also report that the human Fn14 gene is expressed at relatively low levels in normal liver tissue but at high levels in liver cancer cell lines and in hepatocellular carcinoma specimens. Furthermore, the murine Fn14 gene is rapidly induced during liver regeneration in vivo and is expressed at high levels in the hepatocellular carcinoma nodules that develop in the c-myc/transforming growth factor-alpha-driven and the hepatitis B virus X protein-driven transgenic mouse models of hepatocarcinogenesis. These results indicate that Fn14 may play a role in hepatocyte growth control and liver neoplasia.
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Carcinoma Hepatocelular/genética , Regulación de la Expresión Génica , Expresión Génica , Genes Inmediatos-Precoces , Neoplasias Hepáticas/genética , Regeneración Hepática/genética , Proteínas de la Membrana/genética , Receptores del Factor de Necrosis Tumoral , Células 3T3 , Secuencia de Aminoácidos/genética , Animales , Células Cultivadas , Mapeo Cromosómico , ADN Complementario/genética , Humanos , Hígado/metabolismo , Ratones , Ratones Endogámicos , Datos de Secuencia Molecular , ARN Mensajero/metabolismo , Receptor de TWEAKRESUMEN
Polypeptide growth factors promote cell-cycle progression in part by the transcriptional activation of a diverse group of specific genes. We have used an mRNA differential-display approach to identify several fibroblast growth factor (FGF)-1 (acidic FGF)-inducible genes in NIH 3T3 cells. Here we report that one of these genes, called FGF-regulated (FR)-3, is predicted to encode G/T mismatch-binding protein (GTBP), a component of the mammalian DNA mismatch correction system. The murine GTBP gene is transiently expressed after FGF-1 or calf serum treatment, with maximal mRNA levels detected at 12 and 18 h post-stimulation. FGF-1-stimulated NIH 3T3 cells also express an increased amount of GTBP as determined by immunoblot analysis. These results indicate that elevated levels of GTBP may be required during the DNA synthesis phase of the cell cycle for efficient G/T mismatch recognition and repair.
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Reparación del ADN , Proteínas de Unión al ADN/biosíntesis , Factor 1 de Crecimiento de Fibroblastos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Células 3T3 , Secuencia de Aminoácidos , Animales , ADN Complementario , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/genética , Electroforesis en Gel de Poliacrilamida , Humanos , Ratones , Datos de Secuencia Molecular , ARN Mensajero/metabolismo , Análisis de Secuencia de ADNRESUMEN
The effects of linker length on binding affinity and degree of aggregation have been examined in the antifluorescein 4-4-20 and anticarcinoma CC49 single-chain Fvs. Longer linkers in the antifluorescein sFvs have higher affinities for fluorescein and aggregate less. A proteolytically susceptible site between Lys8 and Ser9, in the previously reported 212 linker has been identified. A new linker sequence, 218 (GSTSGSGKPGSGEGSTKG) was designed in which a proline was placed at the C-terminal side of the proteolytic clip site in the 212 linker. The CC49 sFv containing the 218 linker showed reduced aggregation and was found to be more stable to proteolysis in vitro, when compared to the CC49/212 sFv. The CC49 sFv with the longer 218 linker had higher affinity than CC49/212 sFv. An aggregated CC49/212 sFv sample had higher affinity than CC49/218 sFv. The CC49/218 and CC49/212 sFvs had similar blood clearances in mice, while the aggregated CC49/212 sFv remained in circulation significantly longer. In mice bearing LS-174T human colon carcinoma xenografts, the CC49/218 sFv showed higher tumor uptake than the CC49/212 sFv and lower tumor uptake than the aggregated CC49/212 sFv. The higher tumor uptake of the CC49/218 is most likely a result of its higher resistance to proteolysis. The higher affinity and higher tumor uptake of the aggregated CC49/212 sFv are most likely due to the repetitive nature of the TAG-72 antigen and the higher avidity of multivalent aggregates. When the sFvs were radiolabeled with a lutetium-chelate the CC49/218 sFv showed a lower accumulation in the liver and spleen compared to the aggregated CC49/212 sFv.
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Anticuerpos Monoclonales/genética , Antígenos de Neoplasias/inmunología , Glicoproteínas/inmunología , Fragmentos de Inmunoglobulinas/genética , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/metabolismo , Afinidad de Anticuerpos , Antígenos de Neoplasias/metabolismo , Unión Competitiva , Clonación Molecular , Glicoproteínas/metabolismo , Fragmentos de Inmunoglobulinas/metabolismo , Ratones , Ratones Desnudos , Datos de Secuencia Molecular , Radioinmunoensayo , Distribución TisularRESUMEN
The binding of polypeptide growth factors to their appropriate cell surface transmembrane receptors triggers numerous biochemical responses, including the transcriptional activation of specific genes. We have used a differential display approach to identify fibroblast growth factor-1-inducible genes in murine NIH 3T3 cells. Here, we report that the fibroblast growth factor-inducible-14 (Fn14) gene is a growth factor-regulated, immediate-early response gene expressed in a developmental stage- and adult tissue-specific manner in vivo. This gene, located on mouse chromosome 17, is predicted to encode an 129-amino acid type Ia membrane protein with no significant sequence similarity to any known protein. We have used two experimental approaches, direct fluorescence microscopy and immunoprecipitation analysis of biotinylated cell surface proteins, to demonstrate that Fn14 is located on the plasma membrane. To examine the biological consequences of constitutive Fn14 expression, we isolated NIH 3T3 cell lines expressing variable levels of epitope-tagged Fn14 and analyzed their phenotypic properties in vitro. These experiments revealed that Fn14 expression decreased cellular adhesion to the extracellular matrix proteins fibronectin and vitronectin and also reduced serum-stimulated cell growth and migration. These results indicate that Fn14 is a novel plasma membrane-spanning molecule that may play a role in cell-matrix interactions.