RESUMO
Expression of the E6 oncoprotein of human papillomavirus (HPV) 16 in primary human keratinocytes (PHKs) was previously shown to significantly reduce apoptosis. This could be due to increased cell adhesion. Adhesion ability was tested by seeding cells on tissue culture dishes coated with different concentrations of poly(HEME) and determination of the proportion of attached cells. Assays were carried out with PHKs, immortalized human keratinocytes (HaCaT) and human 293T cells. The E6 gene was transduced via retroviral infection or DNA transfection. Results of these assays showed that expression of E6 increased the proportion of cells that attached to poly(HEME). Several HPV16 E6 mutants were also tested in the above assay in 293T cells. These assays showed that the p53 targeting region of E6 is dispensable for this activity. Assays of inhibition of tyrosine kinases by bombesin showed that E6 probably utilizes other pathways to increase cell adhesion.
Assuntos
Papillomavirus Humano 16/metabolismo , Queratinócitos/metabolismo , Proteínas Oncogênicas Virais/metabolismo , Proteínas Repressoras/metabolismo , Sequência de Aminoácidos , Apoptose/efeitos dos fármacos , Benzoquinonas/farmacologia , Bombesina/farmacologia , Adesão Celular/efeitos dos fármacos , Adesão Celular/genética , Adesão Celular/fisiologia , Linhagem Celular , Inibidores Enzimáticos/farmacologia , Quinase 1 de Adesão Focal/metabolismo , Regulação da Expressão Gênica , Hemoglobinas/metabolismo , Papillomavirus Humano 16/genética , Humanos , Queratinócitos/virologia , Lactamas Macrocíclicas/farmacologia , Dados de Sequência Molecular , Mutação , Neurotransmissores/farmacologia , Proteínas Oncogênicas Virais/genética , Proteínas Repressoras/genética , Rifabutina/análogos & derivados , Alinhamento de Sequência , Proteína Supressora de Tumor p53/metabolismoRESUMO
BGC9331 is a rationally designed, specific nonpolyglutamatable thymidylate synthase (TS) inhibitor that is active in gynaecological malignancies. In the light of the sensitivity of human ovarian tumour cell lines to BGC9331 and non-cross resistance to platinum drugs, we studied the combination BGC9331/carboplatin (BCA) in a phase I (PI) pharmacokinetic (PK) and pharmacodynamic (PD) study in platinum pretreated gynaecological malignancies. Patients were >or=18 years or over, with a histologically confirmed gynaecological malignancy, radiological evidence of relapse, and a platinum treatment free interval of at least 6 months. Up to three prior lines of chemotherapy were permitted. Carboplatin (AUC5) and BGC9331 were administered on day 1, and BGC9331 was also given on day 8 of a 21-day cycle. In total, 14 patients were enrolled, and treated with BGC9331 at four dose levels, 40, 65, 85 and 100 mg m-2. The principal grade 3 and 4 haematological toxicity was neutropaenia. The principal nonhaematological toxicities were lethargy and nausea. Dose-limiting toxicities were seen in two patients at 100 mg m-2 BGC9331 (grade 4 neutropaenia>7 days, and grade 4 fatigue>7 days). Plasma BGC9331 was measured by an ELISA that was adapted for use in humans. Carboplatin was assayed by flameless atomic absorption spectrometry. There was no PK interaction between the two drugs. Plasma deoxyuridine was elevated indicating TS inhibition to at least day 12. Antitumour activity was observed in four out of 14 (28%) of patients. In conclusion, the combination of BGC9331 and carboplatin is well tolerated with no significant PK interaction between the two drugs. There is evidence of TS inhibition with the combination. We have demonstrated antitumour activity in platinum pretreated gynaecological malignancy. Further exploration of this combination in this disease is warranted.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/farmacocinética , Carboplatina/efeitos adversos , Carboplatina/farmacocinética , Inibidores Enzimáticos/efeitos adversos , Inibidores Enzimáticos/farmacocinética , Neoplasias dos Genitais Femininos/tratamento farmacológico , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Área Sob a Curva , Carboplatina/uso terapêutico , Inibidores Enzimáticos/uso terapêutico , Feminino , Humanos , Infusões Intravenosas , Dose Máxima Tolerável , Pessoa de Meia-Idade , Neutropenia/induzido quimicamente , Timidilato Sintase/antagonistas & inibidores , Resultado do TratamentoAssuntos
Planejamento em Saúde Comunitária/organização & administração , Participação da Comunidade/métodos , Medicina Baseada em Evidências/organização & administração , Pesquisa sobre Serviços de Saúde/organização & administração , Relações Comunidade-Instituição , Guias como Assunto , Humanos , Avaliação das Necessidades , Avaliação de Resultados em Cuidados de Saúde , Desenvolvimento de Programas , Projetos de Pesquisa/normas , Apoio à Pesquisa como Assunto/organização & administração , Estados Unidos , United States Agency for Healthcare Research and QualityAssuntos
Antibacterianos/uso terapêutico , Raspagem Dentária/normas , Periodontite/terapia , Aplainamento Radicular/normas , Adulto , Amoxicilina/uso terapêutico , Clorexidina/uso terapêutico , Doença Crônica , Terapia Combinada , Raspagem Dentária/métodos , Medicina Baseada em Evidências , Humanos , Metronidazol/uso terapêutico , Minociclina/uso terapêutico , Índice Periodontal , Periodontite/diagnóstico , Periodontite/tratamento farmacológico , Periodontite/microbiologia , Projetos de Pesquisa , Aplainamento Radicular/métodos , Tetraciclina/uso terapêutico , Resultado do TratamentoAssuntos
Bronquiolite/diagnóstico , Bronquiolite/prevenção & controle , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/economia , Anticorpos Monoclonais Humanizados , Antivirais/administração & dosagem , Antivirais/economia , Bronquiolite/tratamento farmacológico , Criança , Pré-Escolar , Análise Custo-Benefício , Medicina Baseada em Evidências , Guias como Assunto , Humanos , Lactente , Recém-Nascido , Palivizumab , Medicamentos para o Sistema Respiratório/uso terapêuticoRESUMO
4-[N-[7-Bromo-2-methyl-4-oxo-3,4-dihydroquinazolin-6-ylmethyl]-N-(prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide (CB30865) is a quinazolin-4-one antitumor agent whose high growth-inhibitory activity (W1L2 IC(50) = 2.8 +/- 0.50 nM) is believed to have a folate-independent locus of action. In addition, CB30865 represents a class of compounds with unique biochemical characteristics such as a delayed, non-phase specific, cell-cycle arrest. The low aqueous solubility of CB30865 prompted a search for more water-soluble analogues for in vivo evaluation of this class of compounds. It was thought that aqueous solubility could be increased by the introduction of amino functionalities at the 2-position of the quinazolin-4-one ring. A variety of compounds (5a-j, 31a-c, 32, and 33) were synthesized in a linear fashion starting from 3-chloro-4-methylaniline. Most of these compounds (e.g., 5a, 5b, 5g) were significantly more water-soluble than CB30865 (636 microM for 5a at pH 6 and 992 microM for 5g at pH 6). In addition, some of them were up to 6-fold more cytotoxic than CB30865 (e.g., for 5a, W1L2 IC(50) = 0.49 +/- 0.24 nM) and retained its novel biochemical characteristics.
Assuntos
Antineoplásicos/síntese química , Quinazolinas/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Divisão Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Concentração Inibidora 50 , Quinazolinas/química , Quinazolinas/farmacologia , Solubilidade , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
Cyclopenta[g]quinazoline-based inhibitors of thymidylate synthase (TS) possess a chiral centre at the 6-position of the molecule. The effect of this chirality on the inhibition of TS was investigated by synthesising compounds 6S-1a-c, 6R-1a-c. It was shown, in particular with the diastereoisomers 6S-1c, 6R-1c, that the inhibitory activity against TS is mainly due to the 6S diastereoisomer rather than the 6R diastereoisomer, which is virtually inactive.
Assuntos
Antagonistas do Ácido Fólico/química , Antagonistas do Ácido Fólico/farmacologia , Timidilato Sintase/antagonistas & inibidores , Bioquímica/métodos , Ciclopentanos/química , Ciclopentanos/farmacologia , Avaliação Pré-Clínica de Medicamentos , Quinazolinas/química , Quinazolinas/farmacologia , Relação Estrutura-AtividadeRESUMO
ZD9331 is a nonpolyglutamatable antifolate inhibitor of thymidylate synthase currently in clinical development. This enzyme is crucial for DNA synthesis and catalyzes the reductive methylation of dUMP to form thymidylate, which is subsequently converted to dTTP. The pharmacokinetics of two curative antitumor doses of ZD9331 administered by either a single i.p. bolus injection (50 mg/kg) or by 24-h s.c. infusion (3 mg/kg) have been measured in a thymidine salvage-incompetent murine lymphoma model (L5178Y) using a sensitive and specific ELISA. To gain an understanding of the relationship between the pharmacokinetics of ZD9331 and antitumor activity perturbations in tumor, dTTP and dUMP concentrations were also determined. After bolus administration, ZD9331 was eliminated from plasma and tissues relatively rapidly, with terminal elimination (lambda(z) 0-24 h) of 4-6 h. Liver concentrations were 8-fold higher than those measured in the plasma. Kidney and lymphoma drug concentrations were similar to those of plasma, although there was evidence of a slower overall elimination of drug at later time points. Steady-state concentrations of ZD9331 were obtained 4-5 h after the start of the 24 h s.c. infusion. At the end of infusion, elimination rates were similar for plasma and tissues (approximately 3.5 h) but appeared to be slower in the tumor at later time points. Liver concentrations were approximately 4-fold higher, and kidney and tumor concentrations were similar to those in the circulation. Depletion of dTTP and elevation in dUMP in the tumor were consistent with inhibition of thymidylate synthase after both administration schedules, although the time for which dTTP was decreased was longer (approximately 24 h) for the infusional route than for the bolus injection (<16 h). The results suggest that antitumor activity is dependent on attaining adequate drug concentrations to affect dTTP pools as well as on the duration of effective drug levels.
Assuntos
Antineoplásicos/farmacocinética , Linfoma/tratamento farmacológico , Núcleosídeo-Fosfato Quinase/antagonistas & inibidores , Quinazolinas/farmacocinética , Animais , Desoxirribonucleotídeos/metabolismo , Modelos Animais de Doenças , Esquema de Medicação , Ensaio de Imunoadsorção Enzimática , Feminino , Bombas de Infusão , Injeções Intraperitoneais , Injeções Subcutâneas , Linfoma/metabolismo , Camundongos , Camundongos Endogâmicos DBA , Quinazolinas/sangue , Fatores de Tempo , Distribuição TecidualRESUMO
Thymidylate synthase (TS) is an important enzyme catalysing the reductive methylation of dUMP to dTMP that is further metabolized to dTTP for DNA synthesis. Loss of viability following TS inhibition occurs as a consequence of depleted dTTP pools and at least in some cell lines, accumulation of dUTP and subsequent misincorporation of uracil into DNA. The expansion in dUTP pools is largely determined by the expression of the pyrophosphatase, dUTPase. Our previous work has shown that following TS inhibition the ability to accumulate dUTP was associated with an earlier growth inhibitory effect. 3 human lung tumour cell lines and HT29 human colon tumour cells transfected with dUTPase have been used to investigate the relationship between loss of viability following TS inhibition and dUTP accumulation. Cell cycle arrest typical of TS inhibition was an early event in all cell lines and occurred irrespective of the ability to accumulate dUTP or p53 function. However, a large expansion of dUTP pools was associated with mature DNA damage (4 h) and an earlier loss of viability following TS inhibition compared to cells in which dUTP pools were not expanded. In A549 cells damage to mature DNA may have been exacerbated by significantly higher activity of the excision repair enzyme, uracil-DNA glycosylase. Consistent with results using different inhibitors of TS, transfection of dUTPase into HT29 cells significantly reduced the cytotoxicity of a 24 h but not 48 h exposure to ZD9331. Although loss of viability can be mediated through dTTP deprivation alone, the uracil misincorporation pathway resulted in an earlier commitment to cell death. The relevance of this latter pathway in the clinical response to TS inhibitors deserves further investigation.
Assuntos
Nucleotídeos de Desoxiuracil/metabolismo , Timidilato Sintase/antagonistas & inibidores , Células Tumorais Cultivadas/patologia , Animais , Antineoplásicos/farmacologia , Western Blotting , Divisão Celular/efeitos dos fármacos , Ensaio de Unidades Formadoras de Colônias , DNA/metabolismo , Dano ao DNA , Feminino , Citometria de Fluxo , Humanos , Pirofosfatases/genética , Pirofosfatases/metabolismo , Quinazolinas/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Timidilato Sintase/genética , Transfecção , Células Tumorais Cultivadas/enzimologiaRESUMO
Altered expression of genes can have phenotypic consequences in cancer development and treatment, developmental abnormalities, and differentiation processes. Here we describe a rapid approach, termed comparative expressed sequence hybridization (CESH), which gives a genome-wide view of relative expression patterns within tissues according to chromosomal location. No prior knowledge of genes or cloning is required, and minimal amounts of tissue can be used. Expression profiles are achieved in a manner similar to the identification of chromosomal imbalances by comparative genomic hybridization analysis. The approach is demonstrated to indicate a chromosomal region that harbors overexpressed genes that may be associated with a drug-resistant phenotype. In addition, known and new regions of differential gene expression in both normal tissues and tumor samples from the soft tissue sarcoma group of rhabdomyosarcoma (RMS) are indicated. These regions included 2p24; overexpression of MYCN at 2p24 was confirmed by quantitative reverse transcription-PCR for all of the alveolar RMS cases and did not necessarily correspond to genomic amplification. Evidence including region specific microarray analysis indicated that overexpression of several genes from a region may be required for detection by CESH. This evidence is consistent with clusters of functionally related genes and mechanisms that affect the expression of a number of genes at a particular genomic location. The distinctive CESH profiles demonstrated in different subtypes of RMS show potential for tumor classification.
Assuntos
Cromossomos Humanos , Expressão Gênica , Genoma , Cromossomos Artificiais de Levedura , Humanos , Hibridização de Ácido Nucleico , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase , Células Tumorais CultivadasAssuntos
Assistência Odontológica para Doentes Crônicos/métodos , Infecções por HIV , Administração dos Cuidados ao Paciente , Infecções Oportunistas Relacionadas com a AIDS/complicações , Infecções Oportunistas Relacionadas com a AIDS/tratamento farmacológico , Antifúngicos/uso terapêutico , Infecções por HIV/complicações , Pesquisa sobre Serviços de Saúde , Humanos , Doenças da Boca/complicações , Doenças da Boca/tratamento farmacológico , Fatores de Risco , Estados UnidosRESUMO
Apolipoprotein AIV (apo AIV) is a satiety protein secreted by the small intestine. We demonstrate for the first time that apo AIV protein and apo AIV mRNA are present in rat hypothalamus, a site intimately involved in the integration of signals for regulation of food intake and energy metabolism. We further characterized the regulation of hypothalamic apo AIV mRNA levels. Food-deprived animals showed a pronounced decrease in gene expression of apo AIV in the hypothalamus, with a concomitant decrease in the jejunum. Refeeding fasted rats with standard laboratory chow for 4 h evokes a significant increase of apo AIV mRNA in jejunum but not in hypothalamus. However, lipid refeeding to the fasted animals restored apo AIV mRNA levels both in hypothalamus and jejunum. Intracerebroventricular administration of apo AIV antiserum not only stimulated feeding, but also decreased apo AIV mRNA level in the hypothalamus. These data further confirm the central role of apo AIV in the regulation of food intake.
Assuntos
Apolipoproteínas A/genética , Regulação da Expressão Gênica/fisiologia , Hipotálamo/fisiologia , Resposta de Saciedade/fisiologia , Transcrição Gênica , Animais , Anticorpos/farmacologia , Apolipoproteínas A/biossíntese , Apolipoproteínas A/fisiologia , Ventrículos Cerebrais , Infusões Parenterais , Jejuno/fisiologia , Masculino , Regiões Promotoras Genéticas , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Análise de Regressão , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Plasma levels of folates and thymidine in mice are about 10-fold higher than in humans and may influence the therapeutic efficacy of thymidylate synthase (TS) inhibitors, such as 5-fluorouracil (5FU) and the antifolates pemetrexed (MTA) and raltitrexed (RTX). Therefore, we tested their therapeutic efficacy in various murine tumor models, grown in mice on a normal and a folate-depleted diet, with high and low thymidine kinase (TK) levels. MTA and RTX were inactive against Colon-26-10 [doubling times gained by treatment; growth delay factor (GDF), 0.5 and 0.3, respectively], whereas 5FU was very active (GDF, >10; complete cures). Colon-26-10/F, grown in mice on a folate-depleted diet, was more sensitive to RTX and MTA (GDF, 2.1 and 1.3, respectively) but not to 5FU (GDF, 1.2); however, leucovorin reversed the effect leading to cures. Folate depletion did not reverse resistance of Colon-26A and Colon-26G (low TK) to MTA and RTX, whereas leucovorin only enhanced the 5FU effect in Colon-26A and Colon-26A/F. Folic acid at 15 mg/kg did not improve the therapeutic efficacy of MTA in folate-deficient mice. The folate-depleted diet decreased the reduced folates in Colon-26A/F and Colon-26-G/F tumors less (4-5-fold; P < 0.01) than in Colon-26-10/F tumors (8-fold; P < 0.001). Folate depletion increased TS levels 2-3-fold in all of the models and TK levels 6-fold (P < 0.01) in Colon-26G/F, explaining the lack of activity of MTA and RTX in Colon-26G/F. In contrast, TK-deficient FM3A/TK tumors were much more sensitive to RTX, MTA, and 5FU than parent FM3A tumors, which have comparable TS levels. The rate of thymidine phosphorylysis varied considerably in all of the tumors without a clear relation to antitumor activity. In conclusion, tumor folates may potentiate (5FU) or protect (antifolates). Murine tumor models should combine low folates and low thymidine rescue to optimize preclinical testing of antifolates.
Assuntos
Antimetabólitos Antineoplásicos/farmacologia , Inibidores Enzimáticos/farmacologia , Antagonistas do Ácido Fólico/farmacologia , Ácido Fólico/metabolismo , Guanina/análogos & derivados , Timidina/metabolismo , Timidilato Sintase/antagonistas & inibidores , Animais , Antimetabólitos Antineoplásicos/administração & dosagem , Divisão Celular/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Dieta , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Feminino , Fluoruracila/administração & dosagem , Fluoruracila/farmacologia , Ácido Fólico/administração & dosagem , Ácido Fólico/sangue , Glutamatos/farmacologia , Guanina/farmacologia , Leucovorina/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Pemetrexede , Quinazolinas/farmacologia , Tiofenos/farmacologia , Timidina/sangue , Timidina Quinase/metabolismo , Timidilato Sintase/metabolismoAssuntos
Cárie Dentária/diagnóstico , Cárie Dentária/prevenção & controle , Medicina Baseada em Evidências , Anti-Infecciosos Locais/administração & dosagem , Cariostáticos/uso terapêutico , Clorexidina/administração & dosagem , Condutividade Elétrica , Eletrodiagnóstico , Fluoretos Tópicos/uso terapêutico , Humanos , Radiografia Dentária Digital , TransiluminaçãoRESUMO
Sorption of metal cations by stream sediments is an important process affecting the movement of released contaminants in the environment. The ability of cations to desorb from one sediment particle and subsequently sorb to another can greatly affect metal transport rates but rates for this process have not been reported. The objective of this study was to determine the rate at which sorbed metals can migrate from contaminated sediment particles to uncontaminated sediment particles as a function of the concentration of the contaminating solution and the duration of the contact with the contaminating solution. Samples of small sediment particles were exposed to solutions containing cobalt, after which they were rinsed and combined with larger uncontaminated sediment particles in the presence of stream water. Initial concentrations of the contaminating solution ranged from 1ng/l to 1000mg/l and exposures to the contaminating solution ranged from 6h to 14 days. The rate of the migration increased with increasing concentrations in the contaminating solution and with decreasing times of exposure to the contaminating solution. Under the conditions of these experiments, the time required for the migration to reach equilibrium was on the order of months or longer. In separate experiments, the kinetics of adsorption and desorption of cobalt were measured as a function of concentration of the contaminating solution. The time required to reach adsorption equilibrium increased with increasing concentration in the contaminating solution. Times to sorption equilibrium were on the order of months. Desorption was much slower than adsorption and, together with intraparticle diffusion, probably controls the rate of migration from contaminated to uncontaminated sediment. The results of this study show that interparticle migration of metal cations can proceed at significant rates that are strongly influenced by the length of time that the metal has been in contact with the sediment.
Assuntos
Sedimentos Geológicos/química , Metais/química , Poluentes Químicos da Água/análise , Adsorção , Cátions/química , Cinética , Fatores de TempoAssuntos
Antibacterianos/uso terapêutico , Parto Obstétrico/métodos , Medicina Baseada em Evidências , Trabalho de Parto Prematuro/diagnóstico , Trabalho de Parto Prematuro/terapia , Obstetrícia/métodos , Tocolíticos/uso terapêutico , Biomarcadores , Feminino , Monitorização Fetal , Serviços de Assistência Domiciliar , Humanos , Trabalho de Parto Prematuro/epidemiologia , Trabalho de Parto Prematuro/etiologia , Valor Preditivo dos Testes , Gravidez , Projetos de Pesquisa/normas , Resultado do Tratamento , Estados Unidos/epidemiologia , United States Agency for Healthcare Research and Quality , Saúde da MulherRESUMO
The thymidylate synthase inhibitor raltitrexed (ZD1694, Tomudex) induces greater intestinal toxicity, manifested as diarrhea and weight loss, in BALB/c than in DBA/2 mice. No convincing pharmacokinetic or pharmacodynamic reason for this strain difference has been established. We have investigated whether this strain difference in response to raltitrexed is related to differential susceptibilities of intestinal mucosae to undergo apoptosis and also whether p53 expression, a critical factor in 5-fluorouracil-induced intestinal apoptosis and toxicity, modulates this response. Ten mg/kg or 100 mg/kg raltitrexed were administered as single or double i.p. injections 24 h apart to BALB/c, DBA/2, and p53-/- mice. Apoptosis, mitosis, and tissue damage were assessed in intestinal epithelium, and animal weight was recorded. BALB/c mice developed diarrhea and weight loss following 100 mg/kg x2 raltitrexed, whereas DBA/2 mice did not. BALB/c mice were more sensitive than DBA/2 to induction of small-intestinal and colonic apoptosis 24 h following 100 mg/kg raltitrexed. Inhibition of mitosis was equivalent in both strains. Both strains showed histopathological damage to the small intestine after 100 mg/kg x2 raltitrexed, but only BALB/c mice demonstrated colonic damage. p53-null mice showed the same level of small intestinal apoptosis as their wild-type counterparts 24 h after 100 mg/kg x1 raltitrexed and also the same levels of intestinal toxicity 3, 5, and 7 days after 100 mg/kg x2 raltitrexed. Thus, BALB/c mice were more susceptible to induction of intestinal apoptosis by raltitrexed than DBA/2 mice and also demonstrated more histopathological damage in the colon correlating with the induction of diarrhea and weight loss. In contrast to 5-fluorouracil, the intestinal apoptosis and toxicity induced by raltitrexed were p53-independent.
Assuntos
Antimetabólitos Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Quinazolinas/toxicidade , Tiofenos/toxicidade , Proteína Supressora de Tumor p53/fisiologia , Animais , Fluoruracila/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos DBA , Mitose/efeitos dos fármacos , Especificidade da EspécieRESUMO
STUDY DESIGN: Randomized controlled study of standard manual therapy given by 31 generalist physicians to 295 patients, in primary care practice. OBJECTIVES: To determine whether training primary care physicians in techniques of limited manual therapy would result in improved outcomes for their patients with acute low back pain. SUMMARY OF BACKGROUND DATA: Controversy continues regarding the benefit of spinal manual therapy and the role of highly trained manual therapists in the care of low back pain. Continuing medical education in manual therapy is frequently offered to generalist physicians, but nothing is known of the value and effectiveness of this training. METHODS: Thirty-one primary care physicians were trained to provide optimal low back care (enhanced care) and a sequence of eight standard manual therapy techniques. Two hundred ninety-five patients were randomized into two treatment groups: enhanced care alone and enhanced care with manual therapy. Main outcome measures included the Roland-Morris functional disability scale measured over time and patient-reported time to functional recovery, time to complete recovery, and satisfaction with care. RESULTS: No differences were found in Roland-Morris scores over time, mean functional days to recovery, days absent from work, or patient satisfaction. More patients receiving manual therapy (21; 14%) had completely recovered after the first visit compared with the control group (8; 6%; P = 0.01). Patients who received more intense manual therapy (four or more maneuvers) had a more rapid return to functional recovery (7.8 days) compared with those who received less intense manual therapy (11.1 days; P = 0.02). CONCLUSION: Limited training in manual therapy techniques offers very modest benefit compared with high-quality (enhanced) care for acute low back pain. Outcomes may have been modified by failure of some participant physicians to undertake the required sequence of maneuvers. Intensity of manual therapy may be a factor in improving patient outcomes and needs further study.
Assuntos
Educação Médica Continuada , Dor Lombar/reabilitação , Manipulação Ortopédica , Manipulação da Coluna , Médicos de Família/educação , Avaliação da Deficiência , Humanos , Dor Lombar/fisiopatologia , Resultado do TratamentoRESUMO
BACKGROUND: We examined clinical outcomes and patient perceptions of back care given by physicians before and after an intensive course of training in back care and limited manual therapy techniques. METHODS: From a prospective observational cohort study of low back pain involving 208 physicians (115 primary care) and their patients and a subsequent clinical trial of treatment of low back pain given by 31 physicians specially trained in manual therapy and enhanced back care, outcome data from the patients of 13 physicians participating in both studies were compared. In the observational study, the 13 physicians cared for 120 patients. In the manual therapy trial (191 patients) a control group of 94 patients received enhanced back care and an intervention group of 97 patients received enhanced back care plus manual therapy. Pearson's chi-square comparisons and linear and Cox proportional hazard modeling were used to examine effects of variables and recovery time. RESULTS: Characteristics of the 13 physicians' patients in the cohort group and the manual therapy trial showed some differences in income, workers' compensation, previous employment, and baseline dysfunction. Both control and intervention patients in the manual therapy trial showed more rapid improvement in functional status over time and greater satisfaction with their care than those in the previous cohort study. However, there was no difference between the studies in patient-reported time to return to performing usual daily activities. CONCLUSIONS: A structured clinical approach to low back care may bring modestly improved clinical outcomes and patient satisfaction.
Assuntos
Educação Médica Continuada , Medicina de Família e Comunidade/educação , Dor Lombar/terapia , Manipulação Ortopédica , Satisfação do Paciente , Doença Aguda , Adulto , Idoso , Estudos de Coortes , Feminino , Humanos , Masculino , Manipulação Ortopédica/psicologia , Pessoa de Meia-Idade , North Carolina , Observação , Estudos Prospectivos , Resultado do TratamentoRESUMO
A new method is presented for the HPLC determination of plasma 2'-deoxyuridine (dUrd). Briefly, 1 ml of human plasma is deproteinised with perchloric acid followed by purification by solid-phase extraction using a non-polar high-capacity polymeric sorbent. The dUrd is separated on a C18 reversed-phase column using a mobile-phase of 0.05% v/v trifluoroacetic acid in water, with a retention time of 8.5 min at a flow-rate of 1.25 ml min(-1). Quantitation is by UV detection at 261 nm using a photodiode array detector. The limit of quantitation is 6 nM with a linear response over the measured range 6-400 nM. Both intra- and inter-day RSD and bias are typically less than 13%. Chromatograms and pharmacodynamic data from a Phase 1 Clinical Trial of a new antifolate drug, ZD9331 are included to illustrate the utility of the method. They show the increase in circulating dUrd as a result of drug inhibition of the target enzyme thymidylate synthase. The method has the significant advantages of ease and simplicity over earlier methods and may be applied to the analysis of other nucleoside species.