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1.
Bone Joint Res ; 5(8): 353-61, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27587787

RESUMO

OBJECTIVES: Tranexamic acid (TXA) is an antifibrinolytic agent used as a blood-sparing technique in total knee arthroplasty (TKA), and is routinely administered by intravenous (IV) or intra-articular (IA) injection. Recently, a novel method of TXA administration, the combined IV and IA application of TXA, has been applied in TKA. However, the scientific evidence of combined administration of TXA in TKA is still meagre. This meta-analysis aimed to investigate the efficacy and safety of combined IV and IA TXA in patients undergoing TKA. MATERIALS AND METHODS: A systematic search was carried out in PubMed, the Cochrane Clinical Trial Register (Issue12 2015), Embase, Web of Science and the Chinese Biomedical Database. Only randomised controlled trials (RCT) evaluating the efficacy and safety of combined use TXA in TKA were identified. Two authors independently identified the eligible studies, extracted data and assessed the methodological quality of included studies. Meta-analysis was conducted using Review Manager 5.3 software. RESULTS: A total of ten RCTs (1143 patients) were included in this study. All the included studies were randomised and the quality of included studies still needed improvement. The results indicated that, compared with either placebo or the single-dose TXA (IV or IA) group, the combination of IV and IA TXA group had significantly less total blood loss, hidden blood loss, total drain output, a lower transfusion rate and a lower drop in haemoglobin level. There were no statistically significant differences in complications such as wound infection and deep vein thrombosis between the combination group and the placebo or single-dose TXA group. CONCLUSIONS: Compared with placebo or the single-dose TXA, the combined use of IV and IA TXA provided significantly better results with respect to all outcomes related to post-operative blood loss without increasing the risk of thromboembolic complications in TKA.Cite this article: Z. F. Yuan, H. Yin, W. P. Ma, D. L. Xing. The combined effect of administration of intravenous and topical tranexamic acid on blood loss and transfusion rate in total knee arthroplasty: combined tranexamic acid for TKA. Bone Joint Res 2016;5:353-361. DOI: 10.1302/2046-3758.58.BJR-2016-0001.R2.

2.
Eur Rev Med Pharmacol Sci ; 19(11): 1951-8, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26125253

RESUMO

OBJECTIVE: Radiofrequency (RF) shrinkage has been widely conducted in clinical practice and the anterior cruciate ligament (ACL) laxity is regarded as one of the indications. However, basic researches regarding the postoperative histological changes were still insufficient. The study aimed to investigate postoperative histological changes of different areas of ACL for further identifying the optimal area for RF shrinkage. MATERIALS AND METHODS: A total of 29 healthy canine (16.5 ± 2.2 kg, 4.1 ± 0.7 years) were recruited, 24 of which were randomly divided into group A and group B. The epiphyseal arrest was confirmed by X-ray examination in all animals. On one canine, an ACL's vascular perfusion model was established by the ink-perfusion method to observe the blood supply of the ACL. The mid-portion of ACL was conducted by RF in group A while the amph-portions of ACL were conducted in group B. Two legs of each canine were sub-divided into fixation group (group A1 and B1) and non-fixation group (group A2 and B2). 8 ACLs were separated from the rest 4 canine. 2 ACLs were sent for the histological examination after RF shrinkage and the rest 6 ACLs were served as blank controls. Masson staining and hematoxylin-eosin (H-E) staining were applied to observe the features of inner fibrous changes of ACL, cell count and vascular density. RESULTS: According to the Masson staining, collagenous tissues were observed in area after RF shrinkage, which was more evident among group B1 than the others. The cellar density in both group A and B was found lower at 12 weeks postoperatively than that at 6 weeks postoperatively (p < 0.05). In addition, the cellar density in B1 group was found higher than that in A1 group at both 6 and 12 weeks postoperatively (p < 0.05). The density of subsynovial vessel in B1 group was found higher than that in A1 group at 6 weeks postoperatively (p < 0.05) and the density of subsynovial vessel in both A1 and B1 groups was found lower at 12 postoperatively weeks than that at 6 weeks postoperatively (p < 0.05). In both A2 and B2 groups, all ACLs were found ruptured at 12 weeks postoperatively. CONCLUSIONS: The postoperative revascularization pattern of RF-treated ACL was permeating from the synovium to the RF-treated areas, and the best area for the RF shrinkage treatment was the amph-portions of the ACL. Moreover, the application of postoperative external fixation to restrict the movement of injured limb was necessary.


Assuntos
Ligamento Cruzado Anterior/efeitos da radiação , Terapia por Radiofrequência , Animais , Ligamento Cruzado Anterior/cirurgia , Cães , Cuidados Pós-Operatórios , Distribuição Aleatória
3.
Eur J Med Chem ; 43(3): 569-76, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18255197

RESUMO

Neuraminidase (NA) is a critical enzyme of the influenza virus and many inhibitors targeting this enzyme are quite efficient anti-influenza agents. In this paper, quantitative structure-activity relationship (QSAR) method was used to predict the activity of different kinds of 46 NA inhibitors. Heuristic method (HM) and radial basis function network (RBFNN) were first used to build linear and nonlinear models, respectively. The prediction results were in agreement with the experimental value. The proposed model is simple and can be extended to other QSAR investigations.


Assuntos
Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Neuraminidase/antagonistas & inibidores , Relação Quantitativa Estrutura-Atividade , Modelos Lineares , Modelos Biológicos , Neuraminidase/química , Neuraminidase/metabolismo
4.
Eur J Med Chem ; 43(1): 43-52, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17459530

RESUMO

Classification models of estrogen receptor-beta ligands were proposed using linear and nonlinear models. The data set was divided into active and inactive classes on the basis of their binding affinities. The two-class problem (active, inactive) was firstly explored by linear classifier approach, linear discriminant analysis (LDA). In order to get a more accurate prediction model, the nonlinear novel machine learning technique, support vectors machine (SVM), was subsequently used to investigate. The heuristic method (HM) was used to pre-select the whole descriptor sets. The model containing eight descriptors founded by SVM, showed better predictive ability than LDA. The accuracy in prediction for the training, test and overall data sets are 92.9%, 85.8% and 91.4% for SVM, 83.1%, 76.1% and 81.9% for LDA, respectively. The results indicate that SVM can be used as a powerful modeling tool for QSAR studies.


Assuntos
Inteligência Artificial , Receptor beta de Estrogênio/metabolismo , Análise Discriminante , Receptor beta de Estrogênio/agonistas , Receptor beta de Estrogênio/antagonistas & inibidores , Concentração Inibidora 50 , Ligantes , Modelos Lineares , Sensibilidade e Especificidade
5.
Ecotoxicol Environ Saf ; 71(3): 731-9, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18067958

RESUMO

Quantitative structure property relationship (QSPR) models for the prediction of human blood:air partition coefficient (log K(blood)) of volatile organic compounds (VOCs) has been developed based on the linear heuristic method (HM) and non-linear radial basis function neural networks (RBFNNs). Molecular descriptors that are calculated from the structures alone were used to represent the characteristics of the compounds. HM was used both to pre-select the whole descriptor sets and to build the linear model. RBFNN was performed to obtain more accurate models. Both the linear and the non-linear models can give very satisfactory prediction results: the correlation coefficient R was 0.964 and 0.979, and the root-mean-square (RMS) error was 0.3303 and 0.2542 for the whole data set, respectively. The prediction result of the non-linear model is better than that obtained by the linear model. In addition, this paper provides an effective method for predicting log K(blood) from its structures and gives some insight into the structural features related to the solubility of VOCs in human blood.


Assuntos
Poluentes Atmosféricos/metabolismo , Compostos Orgânicos Voláteis/metabolismo , Ar , Poluentes Atmosféricos/sangue , Humanos , Modelos Lineares , Modelos Biológicos , Modelos Químicos , Redes Neurais de Computação , Dinâmica não Linear , Relação Quantitativa Estrutura-Atividade , Solubilidade , Compostos Orgânicos Voláteis/sangue
6.
Nat Biotechnol ; 19(7): 673-6, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11433281

RESUMO

RNA quantitation is becoming increasingly important in basic, pharmaceutical, and clinical research. For example, quantitation of viral RNAs can predict disease progression and therapeutic efficacy. Likewise, gene expression analysis of diseased versus normal, or untreated versus treated, tissue can identify relevant biological responses or assess the effects of pharmacological agents. As the focus of the Human Genome Project moves toward gene expression analysis, the field will require a flexible RNA analysis technology that can quantitatively monitor multiple forms of alternatively transcribed and/or processed RNAs (refs 3,4). We have applied the principles of invasive cleavage and engineered an improved 5'-nuclease to develop an isothermal, fluorescence resonance energy transfer (FRET)-based signal amplification method for detecting RNA in both total RNA and cell lysate samples. This detection format, termed the RNA invasive cleavage assay, obviates the need for target amplification or additional enzymatic signal enhancement. In this report, we describe the assay and present data demonstrating its capabilities for sensitive (<100 copies per reaction), specific (discrimination of 95% homologous sequences, 1 in > or =20,000), and quantitative (1.2-fold changes in RNA levels) detection of unamplified RNA in both single- and biplex-reaction formats.


Assuntos
RNA/análise , Espectrometria de Fluorescência/métodos , Sequência de Bases , Biotecnologia/métodos , HIV/metabolismo , Modelos Genéticos , Dados de Sequência Molecular , RNA/metabolismo , RNA Mensageiro/metabolismo , Proteínas Recombinantes/metabolismo , Homologia de Sequência do Ácido Nucleico
7.
Biochemistry ; 39(31): 9523-32, 2000 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-10924149

RESUMO

The invasive signal amplification reaction is a sensitive method for single nucleotide polymorphism detection and quantitative determination of viral load and gene expression. The method requires the adjacent binding of upstream and downstream oligonucleotides to a target nucleic acid (either DNA or RNA) to form a specific substrate for the structure-specific 5' nucleases that cleave the downstream oligonucleotide to generate signal. By running the reaction at an elevated temperature, the downstream oligonucleotide cycles on and off the target leading to multiple cleavage events per target molecule without temperature cycling. We have examined the performance of the FEN1 enzymes from Archaeoglobus fulgidus and Methanococcus jannaschii and the DNA polymerase I homologues from Thermus aquaticus and Thermus thermophilus in the invasive signal amplification reaction. We find that the reaction has a distinct temperature optimum which increases with increasing length of the downstream oligonucleotide. Raising the concentration of either the downstream oligonucleotide or the enzyme increases the reaction rate. When the reaction is configured to cycle the upstream instead of the downstream oligonucleotide, only the FEN1 enzymes can support a high level of cleavage. To investigate the origin of the background signal generated during the invasive reaction, the cleavage rates for several nonspecific substrates that arise during the course of a reaction were measured and compared with the rate of the specific reaction. We find that the different 5' nuclease enzymes display a much greater variability in cleavage rates on the nonspecific substrates than on the specific substrate. The experimental data are compared with a theoretical model of the invasive signal amplification reaction.


Assuntos
Técnicas de Amplificação de Ácido Nucleico , Polimorfismo Genético , Regiões 3' não Traduzidas/química , Regiões 3' não Traduzidas/metabolismo , Regiões 5' não Traduzidas/química , Regiões 5' não Traduzidas/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , DNA Polimerase I/química , DNA Polimerase I/genética , Exodesoxirribonuclease V , Exodesoxirribonucleases/química , Exodesoxirribonucleases/genética , Hidrólise , Cinética , Modelos Químicos , Sondas de Oligonucleotídeos/química , Oligonucleotídeos/química , Oligonucleotídeos/genética , Reação em Cadeia da Polimerase/métodos , Especificidade por Substrato , Temperatura
8.
J Biol Chem ; 275(32): 24693-700, 2000 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-10827184

RESUMO

DNA replication and repair require a specific mechanism to join the 3'- and 5'-ends of two strands to maintain DNA continuity. In order to understand the details of this process, we studied the activity of the 5' nucleases with substrates containing an RNA template strand. By comparing the eubacterial and archaeal 5' nucleases, we show that the polymerase domain of the eubacterial enzymes is critical for the activity of the 5' nuclease domain on RNA containing substrates. Analysis of the activity of chimeric enzymes between the DNA polymerases from Thermus aquaticus (TaqPol) and Thermus thermophilus (TthPol) reveals two regions, in the "thumb" and in the "palm" subdomains, critical for RNA-dependent 5' nuclease activity. There are two critical amino acids in those regions that are responsible for the high activity of TthPol on RNA containing substrates. Mutating glycine 418 and glutamic acid 507 of TaqPol to lysine and glutamine, respectively, increases its RNA-dependent 5' nuclease activity 4-10-fold. Furthermore, the RNA-dependent DNA polymerase activity is controlled by a completely different region of TaqPol and TthPol, and mutations in this region do not affect the 5' nuclease activity. The results presented here suggest a novel substrate binding mode of the eubacterial DNA polymerase enzymes, called a 5' nuclease mode, that is distinct from the polymerizing and editing modes described previously. The application of the enzymes with improved RNA-dependent 5' nuclease activity for RNA detection using the invasive signal amplification assay is discussed.


Assuntos
DNA Polimerase Dirigida por DNA/química , DNA Polimerase Dirigida por DNA/metabolismo , Endodesoxirribonucleases/metabolismo , RNA/metabolismo , Thermus thermophilus/enzimologia , Thermus/enzimologia , Sequência de Aminoácidos , Archaea/enzimologia , Sequência de Bases , Clonagem Molecular , Cristalografia por Raios X , Interleucina-6/genética , Substâncias Macromoleculares , Modelos Moleculares , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Conformação Proteica , Estrutura Quaternária de Proteína , RNA/química , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , Moldes Genéticos
9.
Gene Ther ; 4(6): 593-9, 1997 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-9231076

RESUMO

We have previously shown that a molecule consisting of a fusion of a Ca(2+)-dependent nuclease (from Staphylococcus aureus) to a retroviral coat protein specifies a potent antiviral specific for that retrovirus. Genes specifying such fusion proteins can be delivered to virus-susceptible cells, providing an antiviral gene therapy aimed at limiting virus spread. We report here the results of experiments to vary the nuclease moiety of such fusion proteins. We found that one nuclease. Serratia marcescens nuclease, was extremely toxic to host cells and hence not likely to be useful for therapeutic purposes. A second nuclease, Escherichia coli RNase Hl was found to be nontoxic and highly effective against a murine leukemia virus when it was fused to the leukemia virus coat protein. The fusion protein was enzymatically active and stably expressed, without apparent toxicity to host cells. Reduction in infectious virus output was as high as 97-99%. These studies provide a model system for the development of gene therapeutic agents aimed at combating retroviral infections in vivo.


Assuntos
Terapia Genética/métodos , Vírus da Leucemia Murina/genética , Leucemia Experimental/terapia , Infecções por Retroviridae/terapia , Infecções Tumorais por Vírus/terapia , Animais , Capsídeo , Escherichia coli , Genes gag , Immunoblotting , Proteínas Recombinantes de Fusão , Ribonuclease H , Cultura de Vírus
10.
Biotechniques ; 23(5): 920-6, 1997 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-9383560

RESUMO

Ribonucleases H (RNases H) are enzymes that specifically degrade the RNA of RNA-DNA hybrids. These enzymes are involved in DNA replication, reverse transcription (RT) and antisense oligodeoxyribonucleotide-mediated arrest of translation. One of the most valuable tools for assaying RNase H activity is the renaturation gel assay with which such activities can be detected using purified protein preparations or crude extracts. Radioactive substrates [32P labeled poly(rA)-poly(dT) hybrid] are commonly used with exposure of the gel to X-ray film; this is possible at any time without disturbing the renaturation-degradation process. Here, we describe a method using fluorescent-labeled substrates. RNA-DNA substrates are synthesized by first transcribing DNA with T7 RNA polymerase using Bodipy-TR-14-UTP and the four normal nucleoside triphosphates. The run-off transcript is annealed to a short oligomeric DNA complementary to the 3'-end of the transcript, and the DNA portion of the hybrid is formed by RT. This RNA-DNA is added to the polyacrylamide mixture before polymerization, and SDS-PAGE is performed as usual. After various periods of renaturation, the gel is scanned to detect fluorescent substrate using the red-excited laser of a fluorescence scanner. This fluorescence method has all of the advantages of using radio-labeled substrates and none of its disadvantages, and the sensitivities of the two methods are comparable. In addition, we show that the sensitivity of this procedure can be increased if damaging chemicals remaining in the gel after polymerization are eliminated by simultaneous electrophoresis of the RNase H and a protein with higher mobility.


Assuntos
Corantes Fluorescentes , Ribonuclease H/análise , DNA/metabolismo , Eletroforese em Gel de Poliacrilamida , Escherichia coli/enzimologia , Géis , Muramidase , Plasmídeos , Desnaturação Proteica , RNA/metabolismo , DNA Polimerase Dirigida por RNA/análise , Ribonuclease H/metabolismo , Saccharomyces cerevisiae/enzimologia , Sensibilidade e Especificidade
11.
Genes Cells ; 1(6): 581-93, 1996 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-9078387

RESUMO

BACKGROUND: Reverse transcription, which converts an RNA genome into double-stranded DNA, requires both the polymerase and RNase H activities of reverse transcriptase (RT). In vitro, poorly processive RT dissociates from partially copied RNA-DNA hybrids, that are usually extended by a second RT molecule. Despite similar structures, RNase HI of Escherichia coli can degrade RNA-DNA hybrids that are resistant to RNase H of RT. E. coli RNase HI is used to determine the accessibility to and requirement for RNA-DNA hybrids in reverse transcription in vivo and in vitro. RESULTS: In the presence of E. coli RNase HI, reverse transcription yields incomplete cDNA molecules due to degradation of RNA-DNA hybrids. Delivery of E. coli RNase HI to Ty1 particles via fusion to the capsid protein can reduce retrotransposition by more than 99%, also indicating inhibition of DNA synthesis in vivo. CONCLUSION: Inhibition of both reverse transcription in vitro and retrotransposition in vivo by E. coli RNase HI indicates that the poor processivity of RT exposes RNA-DNA hybrids critical for reverse transcription to degradation. Targeting a cellular RNase H to HIV may help define the site(s) of RNA-DNA hybrids that are susceptible to nonretroviral RNase H and may be useful for gene therapy to inhibit retroviral replication.


Assuntos
Escherichia coli/enzimologia , Vírus da Leucemia Murina/efeitos dos fármacos , Vírus da Leucemia Murina/genética , Retroelementos/efeitos dos fármacos , Ribonuclease H/farmacologia , Animais , Sequência de Bases , Capsídeo/genética , DNA Recombinante/genética , Escherichia coli/genética , Técnicas In Vitro , Camundongos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/farmacologia , Retroelementos/genética , Ribonuclease H/genética , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/genética , Transcrição Gênica/efeitos dos fármacos
12.
J Clin Eng ; 19(4): 310-23, 1994.
Artigo em Inglês | MEDLINE | ID: mdl-10137112

RESUMO

The Division of Biomedical Engineering (DBME), a vital element in the structure of any medical center, provides complete biomedical equipment services at Westchester County Medical Center (WCMC), through a Biomedical Instrumentation Program. Under this program, the DBME assumes direct responsibility for all diagnostic imaging equipment in radiology, radiation medicine and nuclear medicine; and patient care, surgical life support (respiratory care) equipment in critical care units, operating rooms, G.I. (gastro-intestinal) suites, renal center, burn center, emergency rooms, as well as clinical laboratories. In addition, the DBME provides academic and internship programs, research, design, database support, technology planning, and device inspection or evaluation. The DBME is "looking into the future" for a gradual migration of state-of-the-art technology into healthcare.


Assuntos
Engenharia Biomédica/organização & administração , Serviço Hospitalar de Engenharia e Manutenção/organização & administração , Sistemas de Gerenciamento de Base de Dados , Documentação/métodos , Controle de Formulários e Registros , Hospitais com mais de 500 Leitos , Hospitais de Ensino/organização & administração , Capacitação em Serviço/organização & administração , Relações Interdepartamentais , Internato não Médico/organização & administração , Modelos Organizacionais , New York , Desenvolvimento de Programas/métodos , Gestão de Riscos/organização & administração , Avaliação da Tecnologia Biomédica/organização & administração
13.
Bioorg Med Chem ; 2(3): 169-79, 1994 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-7522907

RESUMO

Ribonuclease H (RNase H) is an endonuclease that cleaves only the RNA strand of an RNA-DNA hybrid to produce 5'-phosphate and 3'-hydroxy termini and lacks useful sequence specific recognition properties. A mutant form of the E. coli enzyme has been prepared that is suited for selective chemical modification at a site proximal to the substrate binding region. The chemical derivatization involves the formation of a disulfide linkage to a modified octadeoxyribonucleotide. The conjugate retains only 0.3% of the normal sequence independent RNase H activity demonstrating that substrate recognition can be modulated by a covalent appendage. A beta-globin RNA transcript containing a sequence complementary to that of the octadeoxyribonucleotide was cleaved in a catalytic fashion to two products upon treatment with the conjugate. The selectivity in the phosphodiester bond cleavage mediated by the conjugate was found to be different than that displayed by the nonderivatized enzyme. These results demonstrate the potential of semi-synthetic RNase H conjugates for mechanistic studies and their application as RNA targeted diagnostic or therapeutic agents.


Assuntos
RNA Mensageiro/metabolismo , Ribonuclease H/metabolismo , Sequência de Bases , Sítios de Ligação , Primers do DNA/genética , Escherichia coli/enzimologia , Escherichia coli/genética , Globinas/genética , Técnicas In Vitro , Modelos Moleculares , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , RNA Bacteriano/química , RNA Bacteriano/genética , RNA Bacteriano/metabolismo , RNA Mensageiro/química , RNA Mensageiro/genética , Especificidade por Substrato
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