RESUMEN
Indole-3-carbinol (I3C) is a phytochemical that exhibits growth-inhibitory activity against various cancer cells. However, there are limited studies on the effects of I3C on colon cancer cells. In this study, the growth-inhibitory activity of I3C against the human colorectal carcinoma cell line (LoVo) was examined. The results of the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide, colony formation, and cell counting assays revealed that I3C suppressed the proliferation of LoVo cells. Microscopy and wound-healing analyses revealed that I3C affected the morphology and inhibited the migration of LoVo cells, respectively. I3C induced apoptosis and DNA fragmentation as evidenced by the results of fluorescein isothiocyanate-conjugated annexin V staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling assay, respectively. Additionally, I3C arrested the cell cycle at the G0/G1 phase and enhanced the reactive oxygen species levels. Western blotting analysis revealed that treatment with I3C resulted in the activation of apoptotic proteins, such as poly(ADP-ribose) polymerase, caspase-3, caspase-7, caspase-9, Bax, Bim, and p53 in LoVo cells. These results indicate that I3C induces apoptosis in LoVo cells by upregulating p53, leading to the activation of Bax and caspases. Taken together, I3C exerts cytotoxic effects on LoVo cells by activating apoptosis.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Neoplasias Colorrectales/tratamiento farmacológico , Indoles/farmacología , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/metabolismo , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Neoplasias Colorrectales/metabolismo , Fragmentación del ADN , Humanos , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
The purpose of this study was to translate and culturally adapt the Carpal Tunnel Questionnaire to produce an equivalent Korean version. A total of 53 patients completed the Korean version of the Carpal Tunnel Questionnaire pre-operatively and 3 months after open carpal tunnel release. All 53 also completed the Korean version of the Disabilities of Arm, Shoulder, and Hand questionnaire pre-operatively and 3 months post-operatively. Reliability was measured by determining the test-retest reliability and internal consistency. Test-retest reliability was assessed using intraclass correlation coefficients and paired t-tests, and internal consistency using Cronbach's alpha coefficients. Pearson correlation analysis was carried out on the Korean version of the Carpal Tunnel Questionnaire scores and the Korean version of the Disabilities of Arm, Shoulder, and Hand scores to assess construct validity. Responsiveness was evaluated using effect sizes and standardized response means. The reliability of the Korean version of the Carpal Tunnel Questionnaire was good. The scores in the Korean version of the Disabilities of Arm, Shoulder, and Hand strongly correlated with the scores in the Korean version of the Carpal Tunnel Questionnaire. Standardized response mean and effect size were both large for the Korean version of the Carpal Tunnel Questionnaire. The study shows that the Korean version of the Carpal Tunnel Questionnaire is a reliable, valid and responsive instrument for measuring outcomes in carpal tunnel syndrome.
Asunto(s)
Síndrome del Túnel Carpiano , Encuestas y Cuestionarios , Adulto , Anciano , Anciano de 80 o más Años , Comparación Transcultural , Evaluación de la Discapacidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Evaluación de Resultado en la Atención de Salud , Reproducibilidad de los Resultados , República de CoreaRESUMEN
After the recent success of several river rehabilitation projects including the Cheong-gye river case, a large number of local governments have been promoting their own projects in Korea. Most of the projects are aimed at securing the soundness of aquatic ecosystems according to the guidelines presented by the Korea Ministry of Environment. However, there is no clear guidance for the management goals of water quality and quantity. In this study, we have made an attempt to construct a habitat database (DB) for each domestic freshwater fish species. The fish population, and physical and physicochemical properties of the habitat of 70 domestic freshwater fish species were investigated using field monitoring data. After the statistical processing, the inhabitable range and optimal range of each species were suggested. Furthermore, based on the DB, a decision support system for ecological river restoration and rehabilitation has been developed, and applied for field tests. It became clear that the decision support procedure based on the fish habitat DB is useful in the planning stage of river rehabilitation projects to select the flagship fish, to decide the restoration goals considering their appropriate habitat and to suggest the optimum quantitative combination of each available water resource.
Asunto(s)
Conservación de los Recursos Naturales , Bases de Datos Factuales , Ecosistema , Peces/fisiología , Ríos , Animales , China , Toma de Decisiones , Contaminación del Agua , Calidad del AguaRESUMEN
Isomerization of a closed to open complex of a promoter upon RNA polymerase binding involves base unpairing at the -10 region. After potassium permanganate sensitivity of unpaired thymine residues, we studied base unpairing at the -10 region during isomerization upon RNA polymerase binding at the P1 and P3 promoters of the gal operon. Substitution of adenine by 2-amino purine (2-AP) at the invariable A small middle dotT base pair at the -11 position of P1 and P3 prevented unpairing not only at that position but also at the other downstream positions, suggesting a "master" role of the adenine base at -11 of the template strand in overall base unpairing. 2-AP at -11 did not inhibit the formation of RNA polymerase small middle dotpromoter complex and subsequent isomerization of the polymerase. Substitution of adenine by 2-AP at several other positions did not affect thymine unpairing. Changing the position of the amino group from C6 in adenine to C2 in 2-AP is mutational only at the master switch position, -11.
Asunto(s)
ARN Polimerasas Dirigidas por ADN/metabolismo , Regiones Promotoras Genéticas , Emparejamiento Base , Secuencia de Bases , ADN Bacteriano , Datos de Secuencia Molecular , Permanganato de Potasio/química , Unión Proteica , Transcripción GenéticaRESUMEN
Previous biochemical assays and a structural model of the protein have indicated that the dimer interface of the Hin recombinase is composed of two alpha-helices. To elucidate the structure and function of the helix, amino acids at the N-terminal end of the helix, where the two helices make their most extensive contact, were randomized, and inversion-incompetent mutants were selected. To investigate why the mutants lost their inversion activities, the DNA binding, hix pairing, invertasome formation, and DNA cleavage activities were assayed using in vivo and in vitro methodologies. The results indicated that the mutants could be divided into four classes based on their DNA binding activity. We propose that the alpha-helices might serve to place a DNA binding motif of Hin in the correct spatial relationship to the minor groove of the recombination site. All the mutants except those that failed to bind DNA were able to perform hix pairing and invertasome formation, suggesting that the dimer interface is not involved in either of these processes. The inversion-incompetent phenotype of the binders was caused by the inability of mutants to perform DNA cleavage. The mutants that showed less binding ability than the wild type nevertheless exhibited a wild-type level of hix pairing activity, because the hix pairing activity overcomes the defect in DNA binding. This phenotype of the mutants that are impaired in DNA binding suggests that the binding domains of Hin may mediate Hin-Hin interaction during hix pairing.
Asunto(s)
Proteínas Bacterianas/genética , ADN Nucleotidiltransferasas/genética , Proteínas de Unión al ADN/genética , Recombinación Genética/genética , Proteínas Bacterianas/metabolismo , Sitios de Unión , ADN Nucleotidiltransferasas/metabolismo , Proteínas de Unión al ADN/metabolismo , Dimerización , Modelos Genéticos , Modelos Moleculares , Mutación , Plásmidos/metabolismo , Unión Proteica , Propiedades de SuperficieRESUMEN
Intracerebroventricular kainate treatment in rats induces neuronal cell death, followed by proliferation and hypertrophy of glial cells in the lesioned area. To further understand the activated signal transduction pathways and to get insights into potential target gene activation, the present study aims to elucidate long-term effects on the phosphorylation state of cAMP response element-binding protein (CREB) in the hippocampal formation. One to four weeks after kainate injection, we found high levels of phosphorylated and hence activated CREB (pCREB) in glial cells of the degenerating CA fields. As shown by electron microscopy, pCREB immunoreactivity was present in reactive astrocytes, oligodendrocyte precursor cells and endothelial cells of blood vessels. It is postulated that pCREB could drive the expression of downstream genes in these cells to promote cell proliferation and survival.
Asunto(s)
Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Hipocampo/metabolismo , Neuroglía/metabolismo , Animales , Endotelio Vascular/metabolismo , Endotelio Vascular/patología , Hipocampo/patología , Inmunohistoquímica , Ácido Kaínico , Masculino , Microscopía Electrónica , Neuronas/metabolismo , Neuronas/patología , Fosforilación , Ratas , Ratas WistarRESUMEN
In order to form the catalytic nucleoprotein complex called the invertasome in the Hin-mediated DNA inversion reaction, interactions of the DNA-binding proteins Hin and Fis are required. Assays for these protein-protein interactions have been exploited with protein cross-linkers in vitro. In this study, an in vivo assay system that probes protein-protein interactions was developed. The formation of a DNA loop generated by protein interactions resulted in transcriptional repression of an artificially designed operon, which in turn increased the chance of survival of Escherichia coli host cells in a streptomycin-containing medium. Using this system, we were able to assay the Hin-Hin interaction that results in the pairing of the two recombination sites and protein interactions that result in the formation of the invertasome. This assay system also led us to find that an individual Hin dimer bound on a recombination site can form a stable complex with Fis bound on the recombinational enhancer; this finding has never been observed in in vitro studies. Possible pathways toward the formation of the invertasome are discussed based on the assay results for a previously reported Hin mutant.
Asunto(s)
Proteínas Bacterianas/metabolismo , Inversión Cromosómica , ADN Nucleotidiltransferasas/metabolismo , ADN Bacteriano , Proteínas de Escherichia coli , Sitios de Unión , Proteínas Portadoras/genética , ADN Nucleotidiltransferasas/genética , Proteínas de Unión al ADN/metabolismo , Dimerización , Elementos de Facilitación Genéticos , Escherichia coli , Factor Proteico para Inverción de Estimulación , Factores de Integración del Huésped , MutaciónRESUMEN
Hin recombinase requires negatively supercoiled DNA for an efficient inversion. We have generated positively supercoiled plasmid DNA using reverse gyrase from Sulfolobus shibatae and subjected it to the Hin-mediated inversion reaction. Both Hin and Fis showed the same DNA binding activity regardless of the superhelical handedness of the substrate plasmid. However, inversion activity on positively supercoiled DNA was less than 1% of negatively supercoiled DNA. Assays designed to probe steps in inversion, showed that on positively supercoiled DNA, Hin was able to cleave the recombination sites with the same efficiency shown on negatively supercoiled DNA but was not able to exchange the cleaved DNA. Based on the theoretical differences between positive and negative supercoiling, our data may suggest that unwinding of the double helix at recombination sites is needed after DNA cleavage for strand exchange to occur.
Asunto(s)
Inversión Cromosómica , ADN Nucleotidiltransferasas/metabolismo , ADN-Topoisomerasas de Tipo II/metabolismo , ADN-Topoisomerasas de Tipo I , ADN Superhelicoidal/metabolismo , Plásmidos/metabolismo , Sitios de Unión , ADN Superhelicoidal/química , Desoxirribonucleasas de Localización Especificada Tipo II , Electroforesis en Gel de Agar , Escherichia coli/genética , Plásmidos/química , Mapeo Restrictivo , Sulfolobus/enzimologíaRESUMEN
We consider the role of polarization in the adsorption of Xe in zeolites of type A by direct comparative analysis of the adsorption isotherms, distributions of occupancies, and 129Xe NMR chemical shifts of Xe(n) in cages containing CaxNa12-2x ions per alpha cage (x = 0, 1, 2, 3, 5). We find that the qualitative trends in the adsorption isotherms, and in the progressions of Xe(n) chemical shifts (for n = 0-8 in cages with x = 0, 1 Ca2+ ions and for n = 0-5 in cages with x = 2, 3 Ca2+ ions) upon increasing the level of Ca2+ ion for Na+ ion substitution could only be accounted for by including polarization of the Xe atom by the zeolite framework and its ions. This system, which permits observation of individual Xe(n) peaks and of directly comparable adsorption isotherms in several cage types, provides a good model system for the interpretation of the more general case in which only the overall average 129Xe NMR chemical shift is observed in open network zeolites, arising from free exchange of Xe among cavities of variable occupancy and variable cation distribution.
Asunto(s)
Xenón/química , Zeolitas/química , Espectroscopía de Resonancia Magnética , Método de Montecarlo , Isótopos de XenónRESUMEN
Low level laser therapy (LLLT) has been shown to produce analgesic effects in many clinical applications. The aim of this clinical study was to test the efficacy of LLLT in controlling orthodontic postadjustment pain. Thirty-nine volunteers were selected for this study that used a double-blind design with placebo control. Elastomeric separators were placed at the proximal contacts of one premolar in each quadrant of the dentition to induce orthodontic pain. The tip of a 30 mW gallium-arsenide-aluminium (830 nm) diode laser probe was then placed at the buccal gingiva and directed at the middle third of the root. Three different treatment durations of 15, 30, and 60 seconds and one placebo treatment of 30 seconds were tested within each subject. The study was conducted over 5 days, and the visual analogue scale (VAS) was used to quantify the pain experienced by the subjects before and after laser applications for each day. Analysis of the VAS median scores showed that teeth exposed to laser treatment had lower levels of pain as compared with those with the placebo treatment. However, nonparametric statistical analysis of the data showed that the differences between treatments and placebo within each subject were not statistically significant.
Asunto(s)
Terapia por Láser , Dolor/prevención & control , Técnicas de Movimiento Dental/efectos adversos , Adulto , Aluminio , Analgesia , Arsenicales , Método Doble Ciego , Galio , Humanos , Aparatos Ortodóncicos/efectos adversos , Dimensión del Dolor , Placebos , Goma , Estadística como Asunto , Factores de Tiempo , Técnicas de Movimiento Dental/instrumentaciónRESUMEN
The Hin recombinase exists as a homodimer in solution and binds to its recombination site (hix) as a dimer (Glasgow, A. C., Bruist, M. F., and Simon M. I. (1989) J. Biol. Chem. 264, 10072-10082). Previous mutational and structural studies of related proteins suggested the location of a putative dimerization domain. In order to probe the function of this region of the protein, cysteine residues were introduced at each of the seven positions comprising the domain. Proteins containing cysteine substitutions at positions 101 and 104 were able to form disulfide bonds spontaneously in crude extracts. M101C showed wild type inversion activity only if the cysteine residue was not engaged in a disulfide bond. This mutant, which is cross-linked by a disulfide bond at the dimeric interface, was found to be defective in the DNA cleavage step during inversion. H107C displayed the wild type DNA cleavage activity, even though its DNA binding activity was not detected by three different binding assays. This suggests that it maintains specificity for DNA binding but dissociates rapidly after binding. The remaining inversion-defective mutants fell into two groups. One group lost its capacity to specifically bind to DNA (G102C, F105C, and F106C), and the other group (R103C and F104C), while able to bind to DNA, was defective in subsequent steps of the inversion reaction. Characteristics of these mutants led me to postulate that movement of subunits at the dimerization interface is critical during DNA binding and during formation of the invertasome, which is the recombination structure. Furthermore, the relative position of subunits within the dimer may be important for maintaining stable DNA binding.
Asunto(s)
ADN Nucleotidiltransferasas/metabolismo , Disulfuros/química , Secuencia de Aminoácidos , ADN Nucleotidiltransferasas/química , Proteínas de Unión al ADN/metabolismo , Ditiotreitol/química , Hidrólisis , Cinética , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Oxidación-Reducción , Polímeros , Homología de Secuencia de AminoácidoRESUMEN
We cloned a mouse homolog of sulfated glycoprotein-2 (SGP-2) cDNA by screening a mouse testicular cDNA library and its nucleotide sequence was determined. The predicted amino acid sequence of the cDNA shares 93% identity with that of rat SGP-2. The nucleotide sequences of both cDNAs show extensive homology throughout the open reading frames and 3' untranslated regions. The 5' untranslated regions, however, share homology only up to 28 bp upstream from the start codons; the rest of sequences are quite different. DNA sequence homology search to mouse SGP-2 cDNA through the EMBL/GenBank database and a recent study on the genomic organization of rat TRPM-2 gene suggest a possibility that there are at least two different SGP-2 mRNAs as a result of alternative splicing and/or different promoter usage in mouse.
Asunto(s)
Glicoproteínas/genética , Chaperonas Moleculares , ARN Mensajero/genética , Testículo/química , Empalme Alternativo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Clusterina , Exones/genética , Biblioteca de Genes , Masculino , Ratones , Datos de Secuencia Molecular , Regiones Promotoras Genéticas/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido NucleicoRESUMEN
A series of biochemical assays were developed and performed to monitor the molecular events that occur during the Hin-mediated DNA inversion reaction. These events can be divided into five different stages: 1) binding of proteins (Hin, Fis, and HU) to DNA; 2) pairing of Hin-binding sites; 3) invertasome formation; 4) DNA strand cleavage; 5) strand rotation and religation. A series of topoisomers of the wild type DNA substrate plasmid (ranging from fully relaxed molecules to those with more than the physiological superhelical density (the physiological superhelical density of pKH336 from Escherichia coli DH10B is -0.072 in this study)) was generated, and the role of negative supercoiling in each step of the inversion reaction was investigated. We found differences in the dependence of the formation of paired Hin-binding sites and of the invertasome formation on the superhelical density of the substrate plasmid. Pairing of Hin-binding sites occurs independently from invertasome formation, and a relatively low degree of negative supercoiling is enough to promote maximal pairing. However, efficient invertasome formation requires higher levels of negative supercoiling.
Asunto(s)
ADN Superhelicoidal/genética , Recombinación Genética , Sitios de Unión , Inversión Cromosómica , ADN Nucleotidiltransferasas/metabolismo , Electroforesis en Gel de Agar , Mutagénesis Sitio-Dirigida , PlásmidosRESUMEN
An artificial recombination site hixC composed of two identical half-sites that bind the Hin recombinase served as a better operator in vivo than the wild type site hixL (Hughes, K. T., Youderian, P., and Simon, M. I (1988) Genes & Dev. 2, 937-948). In vitro binding assays such as gel retardation assay and methylation protection assay demonstrated that Hin binds to hixC as tightly as it binds to hixL, even when the sites are located in negatively supercoiled plasmids. However, hixC served as a poor recombination site when it was subjected to the standard inversion assay in vitro. hixC showed a 16-fold slower inversion rate than the wild type. A series of biochemical assays designed to probe different stages of the Hin-mediated inversion reaction, demonstrated that Hin dimers bound to hixC have difficulty in forming paired hix site intermediates. KMnO4 and S1 nuclease assays detected an anomalous structure of the center of hixC only when the site was in negatively supercoiled plasmids. Mutational analysis in the central region of hixC and assays of paired hix site formation with topoisomers of the hixC substrate plasmid suggested that Hin is not able to pair hixC sites because of the presence of the anomalous structure in the center of the site. The structure does not behave like a DNA "cruciform" since Hin dimers still bind efficiently to the site. It is thought to consist of a short denatured "bubble" encompassing 2 base pairs. During the study of mutations in the center of hixC, it was found that Hin is not able to cleave DNA if a guanine residue is one of the two central nucleotides close to the cleavage site. Furthermore, Hin acts in a concerted fashion and cannot cleave any DNA strand if one of the four strands in the inversion intermediate is not cleavable.
Asunto(s)
ADN Nucleotidiltransferasas/metabolismo , Recombinación Genética , Secuencia de Bases , Sitios de Unión , Inversión Cromosómica , ADN Superhelicoidal/genética , ADN Superhelicoidal/metabolismo , Datos de Secuencia Molecular , PlásmidosRESUMEN
An amino acid residue functioning as a general base has been proposed to assist in the hydrolysis of beta-lactam antibiotics by the zinc-containing Bacillus cereus beta-lactamase II [Bicknell & Waley (1985) Biochemistry 24, 6876-6887]. Oligonucleotide-directed mutagenesis of cloned Bacillus cereus 5/B/6 beta-lactamase II was used in an 'in vivo' study to investigate the role of carboxy-group-containing amino acids near the active site of the enzyme. Substitution of asparagine for the wild-type aspartic acid residue at position 81 resulted in fully functional enzyme. An aspartic acid residue at position 90 is essential for beta-lactamase II to confer any detectable ampicillin and cephalosporin C resistance to Escherichia coli. Conversion of Asp90 into Asn90 or Glu90 lead to the synthesis of inactive enzyme, suggesting that the spatial position of the beta-carboxy group of Asp90 is critical for enzyme function.
Asunto(s)
Asparagina , Bacillus cereus/enzimología , Cefalosporinasa/genética , Mutagénesis Sitio-Dirigida , Secuencia de Aminoácidos , Ácido Aspártico , Bacillus cereus/genética , Secuencia de Bases , Sitios de Unión , Western Blotting , Cefalosporinasa/metabolismo , Clonación Molecular , Escherichia coli/genética , Vectores Genéticos , Datos de Secuencia Molecular , Fenotipo , Proteínas Recombinantes/metabolismo , Mapeo RestrictivoRESUMEN
Selected factors have been evaluated in order to determine their influences on the plasma lipoprotein proton NMR spectra of normal and cancer patients. The variables were donor's diet (fasting/non-fasting), temperature and time of sample storage, processing procedure, centrifugation speed, and water pre-saturation time. Plasma samples from fasting individuals that were placed immediately on ice, spun at 1,000 and 3,000 g for 15 minutes, and the proton NMR spectrum acquired with the Carr-Purcell Meiboom-Gill (CPMG) pulse sequence, using a two-second water pre-saturation time, consistently gave reproducible results. Resonances attributed to lactate were minimized under these processing conditions. Centrifugation speed and pre-saturation time did not affect the average line width; however, donor fasting state, processing temperature, and storage time did alter the line width. Most important, blood chemistry analysis revealed an inverse correlation between triglyceride levels and average methyl and methylene line widths. Thus, these factors alone caution against the indiscriminate use of proton NMR spectra to differentiate plasma from normal and cancer patients.
Asunto(s)
Lipoproteínas , Neoplasias/diagnóstico , Adulto , Dieta/efectos adversos , Humanos , Lipoproteínas/sangre , Espectroscopía de Resonancia Magnética , Persona de Mediana Edad , Reproducibilidad de los Resultados , Temperatura , Factores de Tiempo , Triglicéridos/sangreRESUMEN
Random in vitro mutagenesis of a cloned Bacillus cereus 5/B/6 beta-lactamase II gene was used to select defective genes unable to confer ampicillin or cephalosporin C resistance to Escherichia coli. DNA sequencing of mutant genes identified histidine at position 28 as important to beta-lactamase II function. In addition, the isolation of six identical frameshift mutants established that the carboxyl-terminal end of beta-lactamase II is critical for enzyme function. Random mutagenesis also revealed that His88 (implicated previously as one of 4 residues acting as a zinc ligand) is crucial to enzymatic activity and that a glycine to glutamic acid substitution at position 148 produced a defective beta-lactamase. Oligonucleotide mutagenesis directed at Glu37 and Glu212 suggests that these residues are inconsequential to enzyme function but that histidine at position 28 may be involved in substrate binding or recognition.
Asunto(s)
Bacillus cereus/enzimología , Mutación , beta-Lactamasas/genética , Secuencia de Aminoácidos , Resistencia a la Ampicilina/genética , Secuencia de Bases , Catálisis , Cefalosporinas , ADN/análisis , Farmacorresistencia Microbiana/genética , Electroforesis en Gel de Poliacrilamida , Datos de Secuencia Molecular , Plásmidos , Especificidad por SustratoRESUMEN
Seventy-two patients attending this hospital with a diagnosis of lichen planus were examined. There was a striking predominance of Indians (69%) and a surprisingly low incidence of Chinese (24%) and Malays (4%) in the study population, compared to the racial composition of the general clinic population. The age distribution curve of the study population was bimodal with peaks at age groups 21-30 (22%) and 41-60 (33%) years. The most common morphologic variant was lichen planus vulgaris (common type) which we observed in 46 (64%) patients, followed by lichen planus hypertrophicus in 8 (11%) and lichen planus atrophicus in 2 (3%). Lichen planus confined to the skin was observed in 49 (68%) patients. Mucous membrane involvement was seen in 21 (29%)-17 (24%) had oral mucosa membrane involvement and 4 (6%) genital mucous membrane involvement. In 16 (22%) cases, mucous membranes were exclusively involved--12 (17%) had oral mucous membrane involvement and 4 (6%) genital mucous membrane involvement. Nail changes were only observed in 2 (3%) patients. Eight (11%) patients had associated diabetes mellitus. Overall, lichen planus appeared to pursue a protracted course with only 4 (6%) patients clearing completely after a disease duration of 5-12 months.
Asunto(s)
Liquen Plano , Adolescente , Adulto , Factores de Edad , Anciano , Niño , China/etnología , Estudios de Cohortes , Femenino , Humanos , India/etnología , Liquen Plano/epidemiología , Liquen Plano/etnología , Liquen Plano/patología , Masculino , Persona de Mediana Edad , Membrana Mucosa/patología , SingapurRESUMEN
The Rd gene is expressed in the livers and oviducts of laying hens and codes for the riboflavin-binding protein (RfBP) of egg yolk and egg white. A lambda gt11 cDNA library derived from chicken oviduct poly(A)+ RNA was screened with polyclonal rabbit antiserum to chicken RfBP. Positive clones were isolated and rescreened with a mixed oligonucleotide probe corresponding to residues 20-25 of the mature protein. The largest cDNA clone (969 base pairs) was subcloned into plasmid pIBI21, and the nucleotide sequence was determined by the dideoxynucleotide method. This clone contained the entire coding region for RfBP. The published amino acid sequence of the mature protein was confirmed. In addition, the following 17-residue signal peptide was deduced: Met-Leu-Arg-Phe-Ala-Ile-Thr-Leu-Phe-Ala-Val-Ile-Thr-Ser-Ser-Thr-Cys. Unexpectedly, the nucleotide sequence codes for 2 adjacent arginine residues at the carboxyl terminus that are not observed in the mature protein. The amino acid sequence of RfBP is homologous with bovine milk folate-binding protein. Eight of the nine pairs of cysteines involved in disulfide bonds in RfBP are conserved in folate-binding protein, as are all of the tryptophan residues. Sequence identity between homologous regions of these two vitamin-binding proteins is more than 30%.
Asunto(s)
Proteínas Portadoras/genética , ADN/análisis , Proteínas de Transporte de Membrana , Receptores de Superficie Celular , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Proteínas Portadoras/análisis , Bovinos , Pollos , Receptores de Folato Anclados a GPI , Leche/análisis , Datos de Secuencia MolecularRESUMEN
Two forms of heat-stable, zinc-containing beta-lactamase II have been described for strains of Bacillus cereus and have been shown to differ in substrate specificity (R. B. Davies, E. P. Abraham, J. Fleming, and M. R. Pollock, Biochem. J. 145: 409-411, 1975). We report here the nucleotide sequence, inferred amino acid sequence, and expression of beta-lactamase II from B. cereus 5/B/6 and compare our results with those for its homolog characterized in B. cereus 569/H (M. Hussain, C. Anthony, M. J. Madonna, and J. O. Lampen, J. Bacteriol. 164: 223-229, 1985) to document amino acid differences contributing to the specific properties of these enzymes.