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1.
Rev. latinoam. psicol ; 47(2): 124-125, mayo-ago. 2015. ilus
Article in English, Spanish | LILACS, COLNAL | ID: lil-776352

ABSTRACT

El optimismo y la competitividad se presentan como dos variables importantes a estudiar para comprender las características psicológicas asociadas al máximo rendimiento. La presente investigación se propone analizar las diferencias en optimismo y competitividad en función de: practicar deporte o no; el nivel competitivo; la categoría por edad, y el sexo. Para ello, se aplicaron los cuestionarios LOT-R en su versión en español (Otero, Luengo, Romero, Gómez & Castro, 1998) y Competitividad-10 (Remor, 2007) a una muestra de participantes constituida por 148 atletas (75 hombres y 73 mujeres) y 58 no deportistas (22 hombres y 36 mujeres), con edades comprendidas entre los 14 y los 17 años. En cuanto al rasgo optimismo, los resultados indican que los deportistas son más optimistas que los no deportistas, no se obtienen diferencias en función del nivel competitivo, los deportistas de alto rendimiento de mayor edad son más optimistas que los de menos edad y los hombres son más optimistas que las mujeres. En cuanto a la competitividad, los deportistas de alto rendimiento son más competitivos que los deportistas amateur. No se obtienen diferencias en función de la categoría por edad y los hombres son más competitivos que las mujeres. Se concluye que el optimismo se presenta como una variable asociada principalmente con la práctica deportiva, mientras que la competitividad discrimina el nivel competitivo de los deportistas.


Optimism and competitiveness are presented as two important variables to study in order to understand the psychological characteristics associated with the highest performance. This study attempts to analyze the differences in optimism and competitiveness in relation to: whether or not to sport practice, competitive level, age category, and sex. To do this, the Spanish version of LOT-R (Otero, Luengo, Romero, Gómez and Castro, 1998) and Competitivity-10 (Remor, 2007) questionnaires were applied to a sample of 148 athletes (75 men and 73 women) and 58 non-athletes (22 men and 36 women) with ages ranging from 14 to 17 years. As far as the optimism trait is concerned, the results indicate that athletes are more optimistic than non-athletes. There were no differences in relation to the competition level. The older high performance athletes were, the more optimistic they were compared to their younger counterparts, with males being more optimistic than females. With regard to competitiveness, high performance athletes are more competitive than amateur athletes. There were no differences in relation to age category, with males being more competitive than women. It is concluded that optimism is a construct mainly associated with the sport practice, while competitiveness discriminates athlete competitive level.


Subject(s)
Humans , Male , Female , Adolescent , Binding, Competitive , Athletes , Optimism , Adolescent , Athletic Performance
2.
Indian J Exp Biol ; 2015 Feb; 53(2): 67-74
Article in English | IMSEAR | ID: sea-158377

ABSTRACT

The catabolism of fungal 4-aminobutyrate (GABA) occurs via succinic semialdehyde (SSA). Succinic semialdehyde dehydrogenase (SSADH) from the acidogenic fungus Aspergillus niger was purified from GABA grown mycelia to the highest specific activity of 277 nmol min-1 mg-1, using phenyl Sepharose and DEAE Sephacel chromatography. The purified enzyme was specific for its substrates SSA and NAD+. The substrate inhibition observed with SSA was uncompetitive with respect to NAD+. While product inhibition by succinate was not observed, NADH inhibited the enzyme competitively with respect to NAD+ and noncompetitively with respect to SSA. Dead-end inhibition by AMP and p-hydroxybenzaldehyde (pHB) was analyzed. The pHB inhibition was competitive with SSA and uncompetitive with NAD+; AMP competed with NAD+. Consistent with the kinetic data, a sequential, ordered Bi Bi mechanism is proposed for this enzyme.


Subject(s)
Adenosine Monophosphate/metabolism , Adenosine Monophosphate/pharmacology , Aspergillus niger/enzymology , Aspergillus niger/metabolism , Benzaldehydes/metabolism , Benzaldehydes/pharmacology , Binding, Competitive , Biocatalysis/drug effects , Fungal Proteins/isolation & purification , Fungal Proteins/metabolism , Kinetics , Mycelium/enzymology , Mycelium/metabolism , NAD/metabolism , NAD/pharmacology , Protein Binding , Substrate Specificity , Succinate-Semialdehyde Dehydrogenase/isolation & purification , Succinate-Semialdehyde Dehydrogenase/metabolism , gamma-Aminobutyric Acid/analogs & derivatives , gamma-Aminobutyric Acid/metabolism , gamma-Aminobutyric Acid/pharmacology
3.
Braz. j. phys. ther. (Impr.) ; 18(6): 513-520, 09/01/2015. graf
Article in English | LILACS | ID: lil-732354

ABSTRACT

BACKGROUND: Knowing the potential for and limitations of information generated using different evaluation instruments favors the development of more accurate functional diagnoses and therapeutic decision-making. OBJECTIVE: To investigate the relationship between the number of compensatory movements when climbing up and going down stairs, age, functional classification and time taken to perform a tested activity (TA) of going up and down stairs in boys with Duchenne muscular dystrophy (DMD). METHOD: A bank of movies featuring 30 boys with DMD performing functional activities was evaluated. Compensatory movements were assessed using the climbing up and going down stairs domain of the Functional Evaluation Scale for Duchenne Muscular Dystrophy (FES-DMD); age in years; functional classification using the Vignos Scale (VS), and TA using a timer. Statistical analyses were performed using the Spearman correlation test. RESULTS: There is a moderate relationship between the climbing up stairs domain of the FES-DMD and age (r=0.53, p=0.004) and strong relationships with VS (r=0.72, p=0.001) and TA for this task (r=0.83, p<0.001). There were weak relationships between the going down stairs domain of the FES-DMD-going down stairs with age (r=0.40, p=0.032), VS (r=0.65, p=0.002) and TA for this task (r=0.40, p=0.034). CONCLUSION: These findings indicate that the evaluation of compensatory movements used when climbing up stairs can provide more relevant information about the evolution of the disease, although the activity of going down stairs should be investigated, with the aim of enriching guidance and strengthening accident prevention. Data from the FES-DMD, age, VS and TA can be used in a complementary way to formulate functional diagnoses. Longitudinal studies and with broader age groups may supplement this information. .


CONTEXTUALIZAÇÃO: Conhecer as potencialidades e limitações das informações geradas por diferentes instrumentos de avaliação favorece o desenvolvimento mais preciso do diagnóstico funcional e da tomada de decisão terapêutica. OBJETIVO : Investigar a relação entre o número de movimentos compensatórios ao subir e descer escadas, idade, classificação funcional e tempo de realização de atividade (TA) em meninos com Distrofia Muscular de Duchenne (DMD). MÉTODO : Foi utilizado banco de filmes de 30 meninos com DMD realizando atividades funcionais. Os movimentos compensatórios foram avaliados pela Escala de Avaliação Funcional para Distrofia Muscular de Duchenne (FES-DMD), domínio subir e descer escada; a idade, mensurada em anos; a classificação funcional foi pesquisada pela Escala de Vignos (EV), e o TA foi cronometrado. Foi utilizado o teste de correlação de Spearman. RESULTADOS : Existe moderada relação entre a FES-DMD-subir escada e a idade (r=0,53, p=0,004) e forte relação com a EV (r=0,72, p=0,001) e TA dessa tarefa (r=0,83, p<0,001). Houve fraca relação entre a FES-DMD-descer escada e a idade (r=0,40, p=0,032), EV (r=0,65, p=0,002) e o TA dessa tarefa (r=0,40, p=0,034). CONCLUSÃO : Esses achados indicam que a avaliação da tarefa de subir escada pode trazer informações mais relevantes sobre a evolução da doença, embora a atividade de descer escada deva ser pesquisada visando à orientação e prevenção de acidentes. A utilização conjunta de dados provenientes da FES-DMD, da idade e do TA pode se complementar para formulação do diagnóstico funcional. Estudos longitudinais e com outras faixas etárias mais amplas podem complementar tal informação. .


Subject(s)
Humans , Male , Prostatic Hyperplasia/metabolism , Receptors, Androgen/metabolism , Binding, Competitive , Buffers , Charcoal , Cytosol/metabolism , Dextrans , Dihydrotestosterone/metabolism , Electrophoresis, Agar Gel , Enzyme Activation/drug effects , Estrenes/metabolism , Metribolone , Molybdenum/pharmacology , Progesterone/metabolism , Protease Inhibitors/pharmacology , Temperature , Tartrates/pharmacology , Testosterone Congeners/metabolism
4.
Salud colect ; 10(3): 325-337, sep.-dic. 2014. tab
Article in Spanish | LILACS | ID: lil-733293

ABSTRACT

En el marco de la creciente feminización de la profesión médica en México, el artículo indaga sobre las características de este proceso para el caso de la ginecobstetricia. Considerando la feminización como un proceso de cambio, que se analiza cuantitativa y cualitativamente, el artículo se detiene en especial en las experiencias de las mujeres ginecobstetras, experiencias que se dan en el seno de una especialidad que, desde sus orígenes, funcionó como un dispositivo de control del cuerpo de las mujeres. Basado en una investigación etnográfica, el artículo combina fuentes estadísticas, de archivo y de observación de campo. El material que surge de las entrevistas muestra las experiencias y tensiones que viven las ginecobstetras en este contexto.


In the framework of an increasing feminization of the medical profession in Mexico, this article explores the characteristics of this process in the obstetrics and gynecology specialty. Understanding feminization as a process of change to be analyzed both quantitatively and qualitatively, the article focuses special attention on the experiences of female obstetrician-gynecologists within a medical specialty that has since its origins functioned as a mechanism of control over women's bodies. Based on ethnographic research, the article combines statistical and archival sources and field observation. The interviews reveal the experiences and tensions women obstetrician-gynecologists encounter in this context.


Subject(s)
Alcohol Oxidoreductases/chemistry , Alcohol Oxidoreductases/metabolism , Arginine/chemistry , Pseudomonas putida/enzymology , Alcohol Oxidoreductases/genetics , Alcohol Oxidoreductases/isolation & purification , Binding, Competitive/genetics , Catalysis , Enzyme Activation/genetics , Flavin Mononucleotide/metabolism , Kinetics , Ligands , Mutagenesis, Site-Directed , Mandelic Acids/metabolism , Phenylacetates/metabolism , Protein Binding/genetics , Pseudomonas putida/genetics , Substrate Specificity/genetics , Sulfites/metabolism
5.
Indian J Exp Biol ; 2014 Apr; 52(4): 375-382
Article in English | IMSEAR | ID: sea-150369

ABSTRACT

The first set of competitive inhibitors of molt inhibiting hormone (MIH) has been developed using the effective approaches such as Hip-Hop, virtual screening and manual alterations. Moreover, the conserved residues at 71 and 72 positions in the molt inhibiting hormone is known to be significant for selective inhibition of ecdysteroidogenesis; thus, the information from mutation and solution structure were used to generate common pharmacophore features. The geometry of the final six-feature pharmacophore was also found to be consistent with the homology-modeled MIH structures from various other decapod crustaceans. The Hypo-1, comprising six features hypothesis was carefully selected as a best pharmacophore model for virtual screening created on the basis of rank score and cluster processes. The hypothesis was validated and the database was virtually screened using this 3D query and the compounds were then manually altered to enhance the fit value. The hits obtained were further filtered for drug-likeness, which is expressed as physicochemical properties that contribute to favorable ADME/Tox profiles to eliminate the molecules exhibit toxicity and poor pharmacokinetics. In conclusion, the higher fit values of CI-1 (4.6), CI-4 (4.9) and CI-7 (4.2) in conjunction with better pharmacokinetic profile made these molecules practically helpful tool to increase production by accelerating molt in crustaceans. The use of feeding sub-therapeutic dosages of these growth enhancers can be very effectively implemented and certainly turn out to be a vital part of emerging nutritional strategies for economically important crustacean livestock.


Subject(s)
Amino Acid Sequence , Animals , Arthropod Proteins/antagonists & inhibitors , Arthropod Proteins/chemistry , Arthropod Proteins/metabolism , Binding, Competitive , Crustacea/metabolism , Drug Design , Guanylate Cyclase/antagonists & inhibitors , Guanylate Cyclase/chemistry , Guanylate Cyclase/metabolism , Invertebrate Hormones/antagonists & inhibitors , Invertebrate Hormones/chemistry , Invertebrate Hormones/metabolism , Models, Molecular , Molecular Sequence Data , Sequence Homology, Amino Acid
6.
Indian J Biochem Biophys ; 2013 Dec; 50(6): 511-520
Article in English | IMSEAR | ID: sea-150265

ABSTRACT

The study focuses on the importance of Tyr11 amino acid (AA) and subsequent stereochemistry involved in the binding process of neurotensin (NT) with its receptor (NTR)/binding protein(s) as well as the size heterogeneity. Using the binding of 125I-NT with several chicken tissues, it is identified that one of the crucial factors behind all high affinity (Kd ~10 pM) interactions is due to phenolic-OH (Φ-OH) at the para (p) position of Tyr11 within RRPYIL-CO2H (NT8-13) sequence. Replacing the p-OH only in Tyr11 by substituting with p-Cl, p-F and p-NH2 results in significant change of the binding affinity (Kd); p-OH ≈ p-NH2 (~10 pM), p-Cl (~100 pM), p-F (~120 pM). Interestingly, p-NH2 equals to p-OH displaying the highest affinity. Experiments conducted by binding several of the 125I-azido–NT analogs having azido group attached at different positions within the NT molecule have further confirmed the necessity of RRPYIL sequence for high affinity ligand-receptor interaction. The role of Tryp11 in place of Tyr11 in addition to the results above establishes a significant possibility of H–bonding occurring between p-OH of NT and NTR inside the docking space. Photo labeling of the liver tissue by substituted 125I-Y3-azido-NT analogs shows several specifically labeled bands with considerable range of molecular weight (Mr ~90-30 kDa) variations. These results indicate the existence of molecular heterogeneity concerning the sizes of NTR or else any NT binding proteins in the avian tissues. Further, the study has revealed that besides liver, several other chicken tissues also express similar specific high affinity binding (Kd ~20 pM) with varying capacities (Bmax). The order for Bmax is: liver (1.2 pMol/mg) gall bladder (1.03 pMol/mg) > spleen (0.43 pMol/mg) > brain (0.3 pMol/mg) > colon lung (0.15 pMol/mg). In all cases, the binding was reduced by GTPgS (ED50 ~ 0.05 nM), NEM (ED50 ~ 0.50 mM) and NaCl (ED50 ~30 mM), indicating the existence of NTR identical to the mammalian type-1.


Subject(s)
Amino Acid Sequence , Amino Acid Substitution , Animals , Azides/chemistry , Binding, Competitive , Cell Membrane/metabolism , Chickens , Ethylmaleimide/pharmacology , Female , Guanosine 5'-O-(3-Thiotriphosphate) , Liver/cytology , Male , Molecular Weight , Neurotensin/chemistry , Neurotensin/genetics , Neurotensin/metabolism , Protein Binding/drug effects , Pyrazoles/pharmacology , Quinolines/pharmacology , Receptors, Neurotensin/antagonists & inhibitors , Receptors, Neurotensin/chemistry , Receptors, Neurotensin/metabolism , Sodium Chloride/pharmacology , Stereoisomerism , Tyrosine
7.
Article in Chinese | WPRIM | ID: wpr-318711

ABSTRACT

<p><b>OBJECTIVE</b>To determine the plasma protein binding rate of arctiin and arctigenin.</p><p><b>METHOD</b>The ultrafiltration combined with HPLC was employed to determine the plasma protein binding rate of arctiin and arctigenin as well as rat plasma and healthy human plasma proteins.</p><p><b>RESULT</b>The plasma protein binding rate of arctiin with rat plasma at the concentrations of 64. 29, 32.14, 16.07 mg x L(-1) were (71.2 +/- 2.0)%, (73.4 +/- 0.61)%, (78.2 +/- 1.9)%, respectively; while the plasma protein binding rate of arctiin with healthy human plasma at the above concentrations were (64.8 +/- 3.1)%, (64.5 +/- 2.5)%, (77.5 +/- 1.7)%, respectively. The plasma protein binding rate of arctigenin with rat plasma at the concentrations of 77.42, 38.71, 19.36 mg x L(-1) were (96.7 +/- 0.41)%, (96.8 +/- 1.6)%, (97.3 +/- 0.46)%, respectively; while the plasma protein binding rate of arctigenin with normal human plasma at the above concentrations were (94.7 +/- 3.1)%, (96.8 +/- 1.6)%, (97.9 +/- 1.3)%, respectively.</p><p><b>CONCLUSION</b>The binding rate of arctiin with rat plasma protein was moderate, which is slightly higher than the binding rate of arctiin with healthy human plasma protein. The plasma protein binding rates of arctigenin with both rat plasma and healthy human plasma are very high.</p>


Subject(s)
Animals , Binding, Competitive , Blood Proteins , Metabolism , Chromatography, High Pressure Liquid , Furans , Metabolism , Glucosides , Metabolism , Humans , Lignans , Metabolism , Male , Protein Binding , Rats , Rats, Sprague-Dawley , Ultrafiltration , Methods
8.
Chinese Journal of Biotechnology ; (12): 1093-1105, 2012.
Article in Chinese | WPRIM | ID: wpr-342413

ABSTRACT

Protein A and protein G are two well-defined immunoglobulin (Ig)-binding proteins (IBPs), which show affinity for specific sites on Ig of mammalian hosts. Protein A and protein G contained several highly homologous IgG-binding domains which had been demonstrated to have function to bind to IgG. Whether combinations of Ig-binding domains of various IBPs could produce useful novel binding properties remains interesting. We constructed a combinatorial phage library which displayed randomly-rearranged A, B, C, D and E domains of protein A, B2 and B3 domains of protein G. Four rounds molecular evolution of this library directed by all four human IgG subclasses respectively generated a common arrangement of D-C respectively which didn't exist in SpA. The dynamic loss of control phages and increase of the phages displaying two or more binding domains, especially the selective enrichment of D-C and strict selection of its linking peptides demonstrated the efficient molecular evolutions and the significance of the selected D-C arrangement. The phage binding assays confirmed that D-C possessed a binding advantage with four human IgG subclasses compared to SpA. In this work, a novel combination of Ig-binding domains, D-C, was obtained and presented the novel Ig binding properties which provided a novel candidate molecule for the purification, production and detection of IgG antibodies and a new approach for the further study of structures and functions of IBPs.


Subject(s)
Amino Acid Sequence , Antibody Specificity , Bacterial Proteins , Allergy and Immunology , Metabolism , Binding Sites , Binding, Competitive , Evolution, Molecular , Immunoglobulin G , Allergy and Immunology , Metabolism , Molecular Sequence Data , Peptide Library , Sequence Alignment , Staphylococcal Protein A , Allergy and Immunology , Metabolism
9.
Article in Korean | WPRIM | ID: wpr-100065

ABSTRACT

Erlotinib is a low-molecular-weight quinazoline derivative that inhibits the activation of epidermal growth factor receptor (EGFR) tyrosine kinase through competitive binding of the adenosine triphosphate binding domain of the receptor. Patients undergoing anti-EGFR therapy frequently present with cutaneous reactions like a sterile follicular and pustular rash, xerosis, pruritus, paronychia, hair abnormalities and mucositis, which can cause serious discomfort and negatively affect the compliance with anti-EGFR therapy. We report here on an interesting case of hair abnormalities induced by erlotinib (Tarceva(R)) and this presented as eyelash lengthening and hair curling in a 62-year-old woman.


Subject(s)
Adenosine Triphosphate , Binding, Competitive , Compliance , Exanthema , Female , Hair , Humans , Middle Aged , Mucositis , Paronychia , Polyphosphates , Protein-Tyrosine Kinases , Pruritus , Quinazolines , ErbB Receptors , Erlotinib Hydrochloride
10.
Acta Pharmaceutica Sinica ; (12): 165-169, 2011.
Article in Chinese | WPRIM | ID: wpr-348981

ABSTRACT

With IL-6R as target, a new compound 2460A was identified from fungus using HTS screening model. The taxonomics of the produced strain was confirmed to be Trichoderma hazianum rifai after sequencing analysis of rDNA-ITS (internal transcribed spacer). Results showed that this compound has a binding activity on IL-6R competed with IL-6, thus it is a new ligand of IL-6R originating from microbe. With MTT assay, the anti-tumor activities of 2460A were demonstrated on CM126 and HT-29 cell lines separately, the IC50 are 2.17 x 10(-5) mol x L(-1) and 1.8 x 10(-5) mol x L(-1) respectively. The compound affected lightly the HT-29 cell cycle at S phase. Studies for the anti-tumor activity of 2460A in vivo are in progress in our lab.


Subject(s)
Antineoplastic Agents , Metabolism , Pharmacology , Binding, Competitive , Bone Marrow Neoplasms , Pathology , Cell Cycle , Cell Line, Tumor , Cell Proliferation , HT29 Cells , High-Throughput Screening Assays , Humans , Interleukin-6 , Metabolism , Ligands , Receptors, Interleukin-6 , Metabolism , Trichoderma , Chemistry
11.
Protein & Cell ; (12): 320-332, 2011.
Article in English | WPRIM | ID: wpr-757093

ABSTRACT

Proteins of the complement system are known to interact with many charged substances. We recently characterized binding of C1q and factor H to immobilized and liposomal anionic phospholipids. Factor H inhibited C1q binding to anionic phospholipids, suggesting a role for factor H in regulating activation of the complement classical pathway by anionic phospholipids. To extend this finding, we examined interactions of C1q and factor H with lipid A, a well-characterized activator of the classical pathway. We report that C1q and factor H both bind to immobilized lipid A, lipid A liposomes and intact Escherichia coli TG1. Factor H competes with C1q for binding to these targets. Furthermore, increasing the factor H: C1q molar ratio in serum diminished C4b fixation, indicating that factor H diminishes classical pathway activation. The recombinant forms of the Cterminal, globular heads of C1q A, B and C chains bound to lipid A and E. coli in a manner qualitatively similar to native C1q, confirming that C1q interacts with these targets via its globular head region. These observations reinforce our proposal that factor H has an additional complement regulatory role of down-regulating classical pathway activation in response to certain targets. This is distinct from its role as an alternative pathway down-regulator. We suggest that under physiological conditions, factor H may serve as a downregulator of bacterially-driven inflammatory responses, thereby fine-tuning and balancing the inflammatory response in infections with Gram-negative bacteria.


Subject(s)
Binding, Competitive , Allergy and Immunology , Complement Activation , Allergy and Immunology , Complement C1q , Chemistry , Allergy and Immunology , Metabolism , Complement C4b , Complement Factor H , Chemistry , Allergy and Immunology , Metabolism , Complement Pathway, Classical , Allergy and Immunology , Escherichia coli , Allergy and Immunology , Metabolism , Humans , Iodine Radioisotopes , Isotope Labeling , Lipid A , Allergy and Immunology , Metabolism , Liposomes , Allergy and Immunology , Metabolism , Protein Binding , Allergy and Immunology , Recombinant Proteins , Chemistry , Allergy and Immunology , Metabolism , Substrate Specificity
12.
Protein & Cell ; (12): 437-444, 2011.
Article in English | WPRIM | ID: wpr-757079

ABSTRACT

Diverse subtypes of voltage-gated sodium channels (VGSCs) have been found throughout tissues of the brain, muscles and the heart. Neurotoxins extracted from the venom of the Asian scorpion Buthus martensi Karsch (BmK) act as sodium channel-specific modulators and have therefore been widely used to study VGSCs. α-type neurotoxins, named BmK I, BmK αIV and BmK abT, bind to receptor site-3 on VGSCs and can strongly prolong the inactivation phase of VGSCs. In contrast, β-type neurotoxins, named BmK AS, BmK AS-1, BmK IT and BmK IT2, occupy receptor site-4 on VGSCs and can suppress peak currents and hyperpolarize the activation kinetics of sodium channels. Accumulating evidence from binding assays of scorpion neurotoxins on VGSCs, however, indicate that pharmacological sensitivity of VGSC subtypes to different modulators is much more complex than that suggested by the simple α-type and β-type neurotoxin distinction. Exploring the mechanisms of possible dynamic interactions between site 3-/4-specific modulators and region- and/or species-specific subtypes of VGSCs would therefore greatly expand our understanding of the physiological and pharmacological properties of diverse VGSCs. In this review, we discuss the pharmacological and structural diversity of VGSCs as revealed by studies exploring the binding properties and cross-competitive binding of site 3- or site 4-specific modulators in VGSC subtypes in synaptosomes from distinct tissues of diverse species.


Subject(s)
Animals , Binding Sites , Binding, Competitive , Brain , Metabolism , Heart , Physiology , Humans , Insect Proteins , Genetics , Metabolism , Insecta , Ion Channel Gating , Physiology , Kinetics , Mammals , Muscles , Metabolism , Neurotoxins , Chemistry , Classification , Pharmacology , Protein Binding , Scorpions , Chemistry , Sodium , Metabolism , Sodium Channel Blockers , Pharmacology , Sodium Channels , Classification , Genetics , Metabolism , Synaptosomes , Metabolism
13.
Article in Chinese | WPRIM | ID: wpr-355011

ABSTRACT

<p><b>OBJECTIVE</b>To screen and identify the peptides that specifically bind to CD13 on monocytes.</p><p><b>METHODS</b>The phages capable of specific binding to CD13 were screened in the phage-displayed 12-peptide library. The affinity of the selected phages with CD13 was verified with enzyme-linked immunosorbent assay (ELISA). The sequences of the peptides bound to the phages were deduced according to the phage DNA sequences, and the functional peptides aligned using the BLASTP on the Website NCBI were synthesized. To analyze the biological function of the screened peptides, the location of the peptides bound to THP-1 cells was detected using immunofluorescence assay. The blocking effect of WM15 on the peptide binding to THP-1 cells was assessed by immunofluorescence assay.</p><p><b>RESULTS</b>The phages that specifically bound to CD13 were effectively enriched to approach saturation after 4 rounds of panning. The recovery rate in the fourth round was 30 times that in the first round. Twenty selected phages were verified by ELISA, and the signals of 10 phages were higher than the control. The sequences of the peptides P9 and P7 showed 83% and 100% identity with those of human cytomegalovirus (HCMV) UL38 and UL105, respectively. The peptides bound to the cell membrane of THP-1 cells as shown by immunofluorescence assay. The binding of the peptides P9 and P7 to THP-1 cells was blocked by CD13-specific monoclonal antibody WM15 at different levels.</p><p><b>CONCLUSION</b>Two peptides (P7 and P9) that can specifically bind to CD13 have been screened successfully, and these two peptides show specific binding to CD13 on the membrane of THP-1 cells.</p>


Subject(s)
Amino Acid Sequence , Binding, Competitive , CD13 Antigens , Metabolism , Cell Line , Humans , Molecular Sequence Data , Peptide Library , Peptides , Metabolism , Protein Binding
14.
Article in Korean | WPRIM | ID: wpr-200688

ABSTRACT

Avian metapneumovirus (aMPV) causes an acute and highly contagious upper respiratory tract infection in turkeys and chickens. In this study, a competitive ELISA (C-ELISA) was developed for the detection of antibodies to aMPV in chicken sera and/or their egg yolks. This assay is based on the competitive binding of monoclonal antibody with serum antibodies to recombinant aMPV N protein expressed by a recombinant baculovirus. The C-ELISA showed specificity and sensitivity of 100% and 98.0%, respectively, when compared to the virus neutralization test. In specific pathogen-free chickens experimentally infected with aMPV SC1509 strain, the C-ELISA started to detect antibodies to aMPV as early as 5 days post infection from birds infected with aMPV, while a commercial ELISA kit detected first 10 days post infection. The C-ELISA was similar or superior to a commercial ELISA kit when serum and egg yolk samples collected from chickens on six outbreak farms were tested for diagnosis. The C-ELISA developed in the present work provides a short turnaround time and can be a useful diagnostic and screening tool for aMPV infection in the field.


Subject(s)
Antibodies , Baculoviridae , Binding, Competitive , Birds , Chickens , Egg Yolk , Enzyme-Linked Immunosorbent Assay , Mass Screening , Metapneumovirus , Neutralization Tests , Respiratory Tract Infections , Sensitivity and Specificity , Sprains and Strains , Turkeys , Viruses
15.
Protein & Cell ; (12): 563-572, 2010.
Article in English | WPRIM | ID: wpr-757695

ABSTRACT

Surfactant proteins A (SP-A) and D (SP-D), both members of the collectin family, play a well established role in apoptotic cell recognition and clearance. Recent in vitro data show that SP-A and SP-D interact with apoptotic neutrophils in a distinct manner. SP-A and SP-D bind in a Ca(2+)-dependent manner to viable and early apoptotic neutrophils whereas the much greater interaction with late apoptotic neutrophils is Ca(2+)-independent. Cell surface molecules on the apoptotic target cells responsible for these interactions had not been identified and this study was done to find candidate target molecules. Myeloperoxidase (MPO), a specific intracellular defense molecule of neutrophils that becomes exposed on the outside of the cell upon apoptosis, was identified by affinity purification, mass-spectrometry and western blotting as a novel binding molecule for SP-A and SP-D. To confirm its role in recognition, it was shown that purified immobilised MPO binds SP-A and SP-D, and that MPO is surface-exposed on late apoptotic neutrophils. SP-A and SP-D inhibit binding of an anti-MPO monoclonal Ab to late apoptotic cells. Fluorescence microscopy confirmed that anti-MPO mAb and SP-A/SP-D colocalise on late apoptotic neutrophils. Desmoplakin was identified as a further potential ligand for SP-A, and neutrophil defensin as a target for both proteins.


Subject(s)
Apoptosis , Binding, Competitive , Fluorescent Antibody Technique, Indirect , Humans , Neutrophils , Chemistry , Cell Biology , Metabolism , Peroxidase , Metabolism , Protein Binding , Pulmonary Surfactant-Associated Protein A , Metabolism , Pulmonary Surfactant-Associated Protein D , Metabolism
16.
Protein & Cell ; (12): 664-674, 2010.
Article in English | WPRIM | ID: wpr-757422

ABSTRACT

Mannan-binding lectin (MBL) is a soluble innate immune protein that binds to glycosylated targets. MBL acts as an opsonin and activates complement, contributing to the destruction and clearance of infecting microorganisms. Hepatitis C virus (HCV) encodes two envelope glycoproteins E1 and E2, expressed as non-covalent E1/E2 heterodimers in the viral envelope. E1 and E2 are potential ligands for MBL. Here we describe an analysis of the interaction between HCV and MBL using recombinant soluble E2 ectodomain fragment, the full-length E1/E2 heterodimer, expressed in vitro, and assess the effect of this interaction on virus entry. A binding assay using antibody capture of full length E1/E2 heterodimers was used to demonstrate calcium dependent, saturating binding of MBL to HCV glycoproteins. Competition with various saccharides further confirmed that the interaction was via the lectin domain of MBL. MBL binds to E1/E2 representing a broad range of virus genotypes. MBL was shown to neutralize the entry into Huh-7 cells of HCV pseudoparticles (HCVpp) bearing E1/E2 from a wide range of genotypes. HCVpp were neutralized to varying degrees. MBL was also shown to neutralize an authentic cell culture infectious virus, strain JFH-1 (HCVcc). Furthermore, binding of MBL to E1/E2 was able to activate the complement system via MBL-associated serine protease 2. In conclusion, MBL interacts directly with HCV glycoproteins, which are present on the surface of the virion, resulting in neutralization of HCV particles.


Subject(s)
Binding, Competitive , Glycosylation , Hepacivirus , Genetics , Virulence , Physiology , Humans , Mannose-Binding Lectin , Metabolism , Mannose-Binding Protein-Associated Serine Proteases , Metabolism , Monosaccharides , Metabolism , Protein Binding , Protein Multimerization , Tumor Cells, Cultured , Viral Envelope Proteins , Metabolism , Virion , Virulence , Physiology , Virus Internalization
17.
Braz. j. med. biol. res ; 42(3): 299-304, Mar. 2009. ilus, tab, graf
Article in English | LILACS | ID: lil-507340

ABSTRACT

Sleep disturbances have far-reaching effects on the neuroendocrine and immune systems and may be linked to disease manifestation. Sleep deprivation can accelerate the onset of lupus in NZB/NZWF1 mice, an animal model of severe systemic lupus erythematosus. High prolactin (PRL) concentrations are involved in the pathogenesis of systemic lupus erythematosus in human beings, as well as in NZB/NZWF1 mice. We hypothesized that PRL could be involved in the earlier onset of the disease in sleep-deprived NZB/NZWF1 mice. We also investigated its binding to dopaminergic receptors, since PRL secretion is mainly controlled by dopamine. Female NZB/NZWF1 mice aged 9 weeks were deprived of sleep using the multiple platform method. Blood samples were taken for the determination of PRL concentrations and quantitative receptor autoradiography was used to map binding of the tritiated dopaminergic receptor ligands [³H]-SCH23390, [³H]-raclopride and [³H]-WIN35,428 to D1 and D2 dopaminergic receptors and dopamine transporter sites throughout the brain, respectively. Sleep deprivation induced a significant decrease in plasma PRL secretion (2.58 ± 0.95 ng/mL) compared with the control group (25.25 ± 9.18 ng/mL). The binding to D1 and D2 binding sites was not significantly affected by sleep deprivation; however, dopamine transporter binding was significantly increased in subdivisions of the caudate-putamen - posterior (16.52 ± 0.5 vs 14.44 ± 0.6), dorsolateral (18.84 ± 0.7 vs 15.97 ± 0.7) and ventrolateral (24.99 ± 0.5 vs 22.54 ± 0.7 µCi/g), in the sleep-deprived mice when compared to the control group. These results suggest that PRL is not the main mechanism involved in the earlier onset of the disease observed in sleep-deprived NZB/NZWF1 mice and the reduction of PRL concentrations after sleep deprivation may be mediated by modifications in the dopamine transporter sites of the caudate-putamen.


Subject(s)
Animals , Female , Male , Mice , Dopamine Plasma Membrane Transport Proteins/physiology , Lupus Erythematosus, Systemic/etiology , Prolactin/blood , Receptors, Dopamine/physiology , Sleep Deprivation/complications , Autoradiography , Binding, Competitive , Disease Models, Animal , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/metabolism , Mice, Inbred NZB , Sleep Deprivation/metabolism
18.
J Health Popul Nutr ; 2009 Feb; 27(1): 20-30
Article in English | IMSEAR | ID: sea-685

ABSTRACT

The degree of binding of a drug to plasma proteins has a marked effect on its distribution, elimination, and pharmacological effect since only the unbound fraction is available for distribution into extra-vascular space. The protein-binding of atenolol was measured by equilibrium dialysis in the bovine serum albumin (BSA). Free atenolol concentration was increased due to addition of arsenic which reduced the binding of the compounds to BSA. During concurrent administration, arsenic displaced atenolol from its high-affinity binding Site I, and free concentration of atenolol increased from 4.286 +/- 0.629% and 5.953 +/- 0.605% to 82.153 +/- 1.924% and 85.486 +/- 1.158% in absence and presence of Site I probe respectively. Thus, it can be suggested that arsenic displaced atenolol from its binding site resulting in an increase of the free atenolol concentration in plasma.


Subject(s)
Adrenergic beta-Antagonists/chemistry , Animals , Arsenic/chemistry , Atenolol/chemistry , Binding Sites , Binding, Competitive , Biological Availability , Cattle , Drug Interactions , Humans , Serum Albumin/chemistry
19.
Chinese Journal of Biotechnology ; (12): 1042-1048, 2009.
Article in Chinese | WPRIM | ID: wpr-296959

ABSTRACT

We constructed and expressed an anti-CD3/anti-Pgp (P-glycoprotein) diabody previously. However, the two chains of diabody are associated non-covalently, resulting in being capable of dissociating. The aim of this study is to enhance the stability of the diabody. We introduced cysteine residues into the CD3 or Pgp V-domain to covalently lock the two chains together. The disulphide crosslinked diabody were expressed by Escherichia coli (E. coli) 16C9 and purified by a cation exchange column and an anti-Etag affinity chromatography. The purified proteins were verified through SDS-PAGE. Flow cytometry (FCM) was used to analyse the binding properties, competitive binding capacity and stability in vitro. The dsPpg-diabody failed to form disulphide bond properly. The designed disulphide bridge between the different chains of dsCD3-diabody was formed correctly. FCM demonstrated the dsCD3-diabody has specific antigen binding activity, the same binding activity and competitive binding activity as its parent diabody. The dsCD3-diabody retained the full activity even after 72 h incubation at 37 degrees C in human serum, in contrast, the parent diabody began to lose activity after only 1 h and lose all its activity 24 hours later. The induced disulphide bond in the CD3 V-domain effectively enhanced the stability of anti-CD3/anti-Pgp diabody. The method of stabilizing a diabody by introducing a disulphide bond into is practical.


Subject(s)
ATP Binding Cassette Transporter, Subfamily B , Allergy and Immunology , Antibodies, Bispecific , Chemistry , Genetics , Allergy and Immunology , Binding, Competitive , CD3 Complex , Allergy and Immunology , Cell Line , Disulfides , Chemistry , Drug Stability , Escherichia coli , Genetics , Metabolism , Humans
20.
Article in Chinese | WPRIM | ID: wpr-294612

ABSTRACT

The preparation of a novel amphiphilic dietary fiber LDL adsorbent with sulfonic group and laurylamine group was studied. The effects of reaction time and reaction temperature on the adsorption rate were studied. The results show that the adsorption rates for the removal of Total cholesterol (TC), Low-density lipoprotein cholesterol (LDL-C) and High-density lipoprotein cholesterol (HDL-C) are 40.8%, 50.8% and 23.6%, respectively. The amphiphilic adsorbent has better selectivity in removing LDL-C.


Subject(s)
Adsorption , Binding, Competitive , Cholesterol , Blood , Dietary Fiber , Pharmacology , Lipoproteins, LDL , Blood
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