RESUMO
OBJECTIVES: The aim was to assess the efficacy and safety of alprostadil in patients with peripheral arterial occlusive disease (PAOD) Fontaine Stage IV. METHODS: This was a multinational, prospective, randomised, double blind, placebo controlled, parallel group trial. Patients with Stage IV PAOD were equally randomised to either 4 weeks of alprostadil treatment twice daily or to placebo treatment twice daily. The primary efficacy variables were the rate of complete healing of all necrosis and ulceration 12 weeks after the end of treatment and the frequency of major amputations 24 weeks after the end of treatment. RESULTS: A total of 840 patients were randomised between 2004 and 2013. At baseline, no major differences between treatment groups were found. The rate of "complete healing" was 18.4% in patients on alprostadil and 17.2% in patients on placebo. The rates of "major amputations" were 12.6% in patients on alprostadil and 14.6% in patients on placebo. The adjusted difference between alprostadil and placebo including their 95% confidence intervals was 1.1 (-4.0 to 6.3) for "complete healing" and -2.1 (-6.7 to 2.5) for "major amputations." In the subgroup of diabetic patients the rates of major amputations were numerically lower in the alprostadil than placebo group (10.6% vs. 17.4%). Within the total cohort a non-significant difference in "decrease in ulcer area ≥50%" was reached in 30.2% of patients on alprostadil and in 24.3% of patients on placebo at end of treatment. CONCLUSIONS: In this study, superiority of alprostadil over placebo could not be shown. Nevertheless, a slight numerical but not clinically relevant advantage for alprostadil emerged from the "area decrease of ulcers by ≥ 50%," indicating that a healing effect may have started. The results have to be considered in the light of several limitations in study design and conduct.
Assuntos
Alprostadil/uso terapêutico , Doença Arterial Periférica/tratamento farmacológico , Vasodilatadores/uso terapêutico , Idoso , Alprostadil/efeitos adversos , Amputação Cirúrgica , Método Duplo-Cego , Europa (Continente) , Feminino , Humanos , Salvamento de Membro , Masculino , Pessoa de Meia-Idade , Doença Arterial Periférica/diagnóstico , Doença Arterial Periférica/fisiopatologia , Estudos Prospectivos , Fatores de Tempo , Resultado do Tratamento , Grau de Desobstrução Vascular/efeitos dos fármacos , Vasodilatadores/efeitos adversos , Cicatrização/efeitos dos fármacosRESUMO
The effect of several monovalent cations on the Ca2+-induced aggregation and fusion of sonicated phosphatidylserine (PS) vesicles is studied by monitoring the mixing of internal compartments of the fusing vesicles using the Tb/dipicolinic acid assay. The dissociation of the fluorescent Tb-dipicolinate complex which accompanies Ca2+-induced vesicle fusion is determined directly and is due to leakage of contents and entry of medium into vesicles. PS vesicles do not fuse when the medium contains only monovalent cations (at pH 7.4), regardless of the cation concentration or whether there is aggregation of the vesicles. A mass-action kinetic analysis of the data provides estimates for the rate of aggregation, C11, and for the rate of fusion per se, f11. Values of f11 increase dramatically with reduction in monovalent cation concentration and are primarily determined by binding ratios of Ca2+ or Mg2+ per PS. With 300 mM of monovalent cations, the fusion per se is essentially rate-limiting to the overall fusion process and values of f11 are significantly larger with the monovalent cations which bind the least, i.e., according to the sequence tetramethylammonium greater than K+ greater than Na+ greater than Li+. With monovalent cations in concentrations of 100 mM or less, the aggregation is rate-limiting to the fusion and the overall initial fusion rates are determined by an interplay between aggregation and fusion rates. Under conditions of fast aggregation, the Ca2+-induced fusion of small PS vesicles can occur within milliseconds or less.
Assuntos
Cálcio , Lipossomos , Lítio , Magnésio , Fosfatidilserinas , Potássio , Sódio , Animais , Encéfalo , Cátions Monovalentes , Bovinos , Cinética , Modelos BiológicosRESUMO
The influence of cholesterol on divalent cation-induced fusion and isothermal phase transitions of large unilamellar vesicles composed of phosphatidylserine (PS) was investigated. Vesicle fusion was monitored by the terbium/dipicolinic acid assay for the intermixing of internal aqueous contents, in the temperature range 10-40 degrees C. The fusogenic activity of the cations decreases in the sequence Ca2+ greater than Ba2+ greater than Sr2+ much greater than Mg2+ for cholesterol concentrations in the range 20-40 mol%, and at all temperatures. Increasing the cholesterol concentration decreases the initial rate of fusion in the presence of Ca2+ and Ba2+ at 25 degrees C, reaching about 50% of the rate for pure PS at a mole fraction of 0.4. From 10 to 25 degrees C, Mg2+ is ineffective in causing fusion at all cholesterol concentrations. However, at 30 degrees C, Mg2+-induced fusion is observed with vesicles containing cholesterol. At 40 degrees C, Mg2+ induces slow fusion of pure PS vesicles, which is enhanced by the presence of cholesterol. Increasing the temperature also causes a monotonic increase in the rate of fusion induced by Ca2+, Ba2+ and Sr2+. The enhancement of the effect of cholesterol at high temperatures suggests that changes in hydrogen bonding and interbilayer hydration forces may be involved in the modulation of fusion by cholesterol. The phase behavior of PS/cholesterol membranes in the presence of Na+ and divalent cations was studied by differential scanning calorimetry. The temperature of the gel-liquid crystalline transition (Tm) in Na+ is lowered as the cholesterol content is increased, and the endotherm is broadened. Addition of divalent cations shifts the Tm upward, with a sequence of effectiveness Ba2+ greater than Sr2+ greater than Mg2+. The Tm of these complexes decreases as the cholesterol content is increased. Although the transition is not detectable for cholesterol concentrations of 40 and 50 mol% in the presence of Na+, Sr2+ or Mg2+, the addition of Ba2+ reveals endotherms with Tm progressively lower than that observed at 30 mol%. Although the presence of cholesterol appears to induce an isothermal gel-liquid crystalline transition by decreasing the Tm, this change in membrane fluidity does not enhance the rate of fusion, but rather decreases it. The effect of cholesterol on the fusion of PS/phosphatidylethanolamine (PE) vesicles was investigated by utilizing a resonance energy transfer assay for lipid mixing. The initial rate of fusion of PS/PE and PS/PE/cholesterol vesicles is saturated at high Mg2+ concentrations. With Ca2+, saturation is not observed for cholesterol-containing vesicles.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Cátions Bivalentes/farmacologia , Colesterol/farmacologia , Fusão de Membrana/efeitos dos fármacos , Lipídeos de Membrana , Fosfolipídeos , Animais , Varredura Diferencial de Calorimetria , Bovinos , Fenômenos Químicos , Físico-Química , Técnicas In Vitro , Cinética , Fosfatidiletanolaminas , Fosfatidilserinas , TemperaturaRESUMO
We have used the binding of liposomes conjugated with antifluorescein antibody specific for fluorescein isothiocyanate-modified erythrocytes as a model for multivalent antigen-antibody interactions. We examined a series of liposome preparations which were conjugated to between 0 and 332 active antibodies per liposome. The antigen binding capacity and mean intrinsic affinity of the soluble and conjugated antibody were determined by fluorescence quenching of carboxyfluorescein. Liposome-cell interaction data were fitted with a Scatchard-type equation. Functional affinity of liposomes for cells was up to 1000-fold greater than the intrinsic affinity of the antibody for soluble ligand. Analysis for binding at high cell concentrations revealed that liposome-induced cell agglutination reduces the number of available binding sites per cell.
Assuntos
Anticorpos/administração & dosagem , Fluoresceínas , Corantes Fluorescentes , Lipossomos/administração & dosagem , Tiocianatos , Animais , Complexo Antígeno-Anticorpo/análise , Bovinos , Eritrócitos/imunologia , Eritrócitos/metabolismo , Fluoresceína-5-Isotiocianato , Fluoresceínas/metabolismo , Corantes Fluorescentes/metabolismo , Tiocianatos/metabolismoRESUMO
The interaction of La2+ with phosphatidylserine vesicles is studied by differential scanning calorimetry, 140La binding, 31P-NMR chemical shifts and relaxation rates, carboxyfluorescein and [14C]sucrose release, X-ray diffraction and freeze-fracture electron microscopy. In the presence of La3+ concentrations above 1 mM and an incubation temperature of 38 degrees C, i.e., at the phase transition temperature of the complex La/phosphatidylserine, the binding ratio of La/lipid exceeds a 1/3 ratio, reaching saturation at a 1/2 ratio. Analysis, employing a modified Gouy-Chapman equation, indicates a significant increase in the intrinsic binding constant of La/phosphatidylserine when the La3+ concentrations exceeds the threshold concentration for leakage. The analysis illustrates that at the molecular level the binding of La3+ can be comparable to or even weaker than that of Ca2+, but that even when present at smaller concentrations La3+ competes with and partially displaces Ca2+ from membranes or other negatively charged surfaces. The results suggest that the sequence La3+ greater than Ca2+ greater than Mg2+ reflects both the binding strength of these cations to phosphatidylserine as well as their ability to induce leakage, enhancement of 31P spin-lattice relaxation rates, fusion and other structural changes. The leakage, fusion, and other structural changes are more pronounced at the phase transition temperature of the La/lipid complex.
Assuntos
Lantânio/metabolismo , Membranas Artificiais , Fosfatidilserinas/metabolismo , Espectroscopia de Ressonância de Spin Eletrônica , Técnica de Fratura por Congelamento , Lipídeos de Membrana , Métodos , Microscopia Eletrônica , Conformação Molecular , Radioisótopos , Sacarose/metabolismo , Difração de Raios XRESUMO
BACKGROUND: Endoscopic ultrasound (EUS) is an important new tool in the staging of pancreatic malignancies. Using new curved linear-array instruments, real-time fine-needle aspiration biopsy (RTFNA) of pancreatic lesions can be performed. METHODS: Forty-five patients with pancreatic lesions (22 males and 23 females) underwent staging with the Olympus EUM-20 (Olympus America Corp, Melville, NY) followed by EUS-RTFNA with the Pentax FG-32PUA (Pentax-Precision Instrument Corp, Orangeburg, NY) and the 22-gauge GIP needle (GIP Medizin Technik, Grassau, Germany). RESULTS: EUS tumor stages were as follows: TO, n = 1; T1, n = 8; T2, n = 9; and T3 n = 27. Aspiration attempts were unsuccessful in four patients (two technical failures and two inadequate specimens). The remaining 41 lesions (mean size, 3.3 cm) were aspirated under EUS guidance (median passes, three) and the cytologic diagnoses were 25 definite adenocarcinoma, five suspicious for adenocarcinoma (three subsequently confirmed and two clinical course consistent with adenocarcinoma), and 11 negative for malignancy. Of 11 negatives, two were found to have adenocarcinoma, seven were confirmed benign at surgery (four cystadenomas and three inflammatory), one had a benign pseudocyst, and one had abundant inflammatory cells on RTFNA and follow-up time greater than 12 months with computed tomographic (CT) scans consistent with resolving inflammation. There were no false-positive RTFNAs. There were no procedure-related complications. Among those with diagnostic EUS-RTFNA (91%), the sensitivity for malignancy (confirmed plus suspicious) was 94% and negative predictive value 82%. CONCLUSION: EUS-guided RTFNA is a safe and accurate method for performing pancreatic biopsy. It should be considered in patients with suspected pancreatic malignancies in whom a tissue diagnosis is required or when other modalities have failed. EUS-RTFNA allows for local staging and tissue diagnosis in one procedure.
Assuntos
Biópsia por Agulha/métodos , Neoplasias Pancreáticas/diagnóstico , Diagnóstico Diferencial , Endoscopia do Sistema Digestório , Feminino , Humanos , Masculino , Pancreatopatias/diagnóstico , Neoplasias Pancreáticas/diagnóstico por imagem , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/secundário , UltrassonografiaRESUMO
The recent finding that more than one Influenza hemagglutinin (HA) is required at the fusion site for HA-expressing fibroblasts, together with the crystal structure of HA at neutral pH, provide the basic elements of a plausible model for this fusion site. Within an aggregate of HA trimers at low pH, we propose fusion intermediates which are based upon a minimal alteration to the known neutral pH structure of HA and which should have reasonable activation energies. This is the first model of a glycoprotein-mediated fusion site which explicitly accounts for the disposition of the lipids within these intermediates. While the fusion site created by HA will not be the same as that of eukaryotic fusion complexes, general characteristics could be shared.
Assuntos
Hemaglutininas Virais , Fusão de Membrana , Modelos Estruturais , Proteínas do Envelope Viral/metabolismo , Animais , Linhagem Celular , Membrana Celular/fisiologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza , Proteínas do Envelope Viral/químicaRESUMO
Interpretation of soft tissue and bone lesions with fine-needle aspiration (FNA) is difficult. We determined whether clinical history or experience improves diagnostic accuracy of FNA interpretation of these lesions. Four cytopathologists with varying degrees of experience retrospectively reviewed 89 soft tissue and bone lesions, initially without knowledge of clinical history and subsequently with knowledge of clinical history. Each time, the pathologist rendered a precise diagnosis (histogenetic classification, e.g., angiosarcoma, lipoma) for each case and classified the lesion into one of 4 categories: benign, probably benign, probably malignant, or malignant. The proportion of correct precise diagnoses was calculated. Also, the numbers of correct and incorrect classifications of malignancy were used in standard relative operating characteristic (ROC) curve analysis. The area under the ROC curve was estimated to give an indicator of diagnostic accuracy. Knowledge of clinical history increased the proportion of correct diagnoses and the accuracy of the classification for all observers. Experienced observers more accurately classified lesions, and knowledge of clinical history particularly improved the diagnostic accuracy of less experienced observers.
Assuntos
Biópsia por Agulha/normas , Neoplasias Ósseas/patologia , Neoplasias de Tecidos Moles/patologia , Humanos , Prontuários Médicos , Curva ROC , Estudos RetrospectivosRESUMO
Measurements of either HER2 gene overexpression or its gene-coded protein (p185) are clinically useful for predicting prognosis in breast cancer. The measurements are also useful for identifying metastatic breast cancer patients who may benefit from Herceptin treatment. Since fine needle aspiration (FNA) of the breast has become an increasingly popular technique for obtaining tissue specimens, we have developed a sensitive method to quantify p185 in the aspirate. For this procedure, p185 from the cell pellet of FNA is extracted with a buffer containing Triton X-100, and the p185 is measured with an enzyme immunoassay. Most of the malignant breast tumors (N=7) in this study were associated with elevated p185 concentrations (6/7, 319+/-222 U/mg), compared to the p185 concentrations in normal breast tissue (42.8+/-35 U/mg, N=47) or benign lesions (43.1+/-20.2 U/mg, N=22). Quantification of p185 in FNA may improve the assessment of breast cancer patients, revealing whether they are at high risk and may benefit from Herceptin treatment.
Assuntos
Neoplasias da Mama/patologia , Doença da Mama Fibrocística/patologia , Receptor ErbB-2/análise , Biópsia por Agulha , Mama/química , Mama/patologia , Neoplasias da Mama/química , Cromatografia , Diagnóstico Diferencial , Feminino , Doença da Mama Fibrocística/química , Humanos , Valor Preditivo dos Testes , Kit de Reagentes para DiagnósticoRESUMO
Injection of high concentrations of benzo[a]pyrene (10-125 micrograms BaP/g body weight) into young frogs (Xenopus laevis), fish (Gambusia affinis) or tissue culture cells (L 5178 Y) in a state of logarithmic growth causes alterations in DNA. In frogs these alterations reach a maximum at 60-90 min after application, then they decrease and become undetectable after 3 h. Within 8-12 days after the single BaP dose, a new wave of DNA alterations can again be detected. Parallel measurements of mixed function oxygenases (MFO) showed no short term activity changes within the first day. There was, however, an increase of activity starting on the 8th day following injection in frogs, and lasting until day 14. DNA alterations have been assessed by two methods. One is a modification of the alkaline filter elution method of Kohn et al. (1976) and is believed to determine the number of events leading to single strand breaks at alkaline pH. The other method determines the number of S1 nuclease sensitive sites in highly purified native DNA. Both results were found to be highly correlated. The effect does not seem to be caused by an impurity, unless an unknown, very minor, and extraordinarily active component is present. The effect in frogs is linked to nutritional status. Frogs fed a carbohydrate-rich, protein-poor diet do not give the fast response. The fast effect cannot be blocked by application of Actinomycin, Cordycepin, and Chloramphenicol injected either 30 min before or simultaneously with the BaP. Benzo(e)pyrene, a non-tumorigenic isomer of BaP, does not cause any effects at similar doses.
Assuntos
Benzopirenos/toxicidade , Carcinógenos , DNA , Animais , Benzo(a)pireno , Benzopirenos/metabolismo , Peixes , Oxigenases de Função Mista/análise , Xenopus laevisRESUMO
New technologies are trying to make significant changes in the way we perform the Pap smear test, a test that has undergone no significant modification since its inception. Efforts have focused on improving sampling with liquid-based methods, improving detection of abnormal cells with automated screening and improving the detection of human papillomavirus with assays practical for use in routine clinical material. This article reviews the performance of the new technologies to date, with an emphasis on those that are currently approved for cervical cytology testing. In the aggregate, these technologies can have a positive impact on cervical disease detection and intervention.
Assuntos
Carcinoma/patologia , Teste de Papanicolaou , Neoplasias do Colo do Útero/patologia , Esfregaço Vaginal/métodos , Automação , Carcinoma/diagnóstico , Biologia Celular/tendências , Feminino , Humanos , Ciência de Laboratório Médico/tendências , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/patologia , Infecções Tumorais por Vírus/diagnóstico , Infecções Tumorais por Vírus/patologia , Neoplasias do Colo do Útero/diagnóstico , Esfregaço Vaginal/normas , Esfregaço Vaginal/tendênciasRESUMO
Federally-mandated quality control (QC) in Papanicolaou (Pap) smear testing requires rescreening of 10% of negative smears, to include cases selected randomly as well as smears from patients that may have a higher risk for developing cervical cancer based on clinical information. FDA approval of NeoPath's AutoPap 300 QC system (NeoPath, Inc., Redmond, WA) allows practical QC rescreening of all negatives. We tested the ability of AutoPap to help increase identification of detection errors compared to random 10%/high-risk selection. From March 1-August 30, 1997, we utilized AutoPap/high-risk status to select cases for manual rescreen, and compared the rate of identification of primary screening errors to that for the preceding year using 10% random selection/high-risk status. Of 35,027 smears accessioned, 31,240 (89.1%) were screened as negative and 7,965 were selected for manual rescreen. Of these, 353 were determined to be abnormal. Most abnormals identified by this protocol were classified as atypical squamous or glandular cells of undetermined significance (ASCUS or AGUS). However, 59 low-grade squamous intraepithelial lesions (LSIL) and 13 high-grade squamous intraepithelial lesions (HSIL), many with few abnormal cells, were also identified. These results represented an increase in pickup rate of false negative due to detection errors of 2.3-, 2.8- and 5.6-fold for atypical squamous or glandular cells of undetermined significance, LSIL, and HSIL, respectively, when accounting for the volume differences over the time period measured. Our findings strongly support the conclusions drawn from clinical trials of the AutoPap that false negatives due to detection error can be significantly reduced when using AutoPap as part of a routine quality control program.
Assuntos
Processamento de Imagem Assistida por Computador , Programas de Rastreamento/métodos , Teste de Papanicolaou , Garantia da Qualidade dos Cuidados de Saúde/métodos , Neoplasias do Colo do Útero/diagnóstico , Esfregaço Vaginal , Reações Falso-Negativas , Feminino , Humanos , Programas de Rastreamento/instrumentação , Programas de Rastreamento/normas , Garantia da Qualidade dos Cuidados de Saúde/normas , Controle de Qualidade , Esfregaço Vaginal/instrumentaçãoRESUMO
The cell block preparation, which increases cellular yield by capturing any small tissue fragments in a fluid specimen, has been used to define the histologic pattern of the cells in question. This report describes a case in which processing a cell block on residual ThinPrep Pap Test material, using a thrombin-based technique, was useful in augmenting the diagnosis of a glandular lesion.
Assuntos
Técnicas Citológicas , Manejo de Espécimes/métodos , Adenocarcinoma/diagnóstico , Adulto , Carcinoma in Situ/diagnóstico , Núcleo Celular/patologia , Feminino , Humanos , Trombina , Neoplasias do Colo do Útero/diagnóstico , Esfregaço VaginalRESUMO
The acid-fast stain is commonly used in the rapid cytologic assessment of bronchoalveolar lavage (BAL) fluid to detect pulmonary mycobacterial infections, particularly in immunocompromised patients. The identification of acid-fast, rod-shaped organisms may be taken as presumptive evidence of such an infection, in the appropriate clinical setting. However, this determination is made less specific by the occasional acid-fast positivity of microorganisms other than mycobacteria. We report on the occurrence of a fatal pneumonia caused by acid-fast positive Legionella pneumophila detected by BAL. This is a potential pitfall in the rapid diagnosis of pulmonary mycobacterial infections.
Assuntos
Legionella pneumophila/isolamento & purificação , Doença dos Legionários/diagnóstico , Infecções por Mycobacterium/diagnóstico , Pneumonia Bacteriana/diagnóstico , Líquido da Lavagem Broncoalveolar/microbiologia , Citodiagnóstico , Diagnóstico Diferencial , Humanos , Doença dos Legionários/complicações , Masculino , Pessoa de Meia-Idade , Neoplasias da Próstata/complicações , Coloração e RotulagemRESUMO
Diagnosis of lesions of the gastrointestinal tract and adjacent structures is possible using an imaging modality, endoscopic ultrasonography (EUS). Fine-needle aspiration (FNA) is a suitable and cost-effective technique for obtaining cytohistologic material to confirm the diagnosis. EUS is capable of both characterizing the lesion and then guiding the FNA under real-time (RT) ultrasound guidance using a through-the-scope needle aspiration system. The goal of this study was to determine the diagnostic accuracy of this technique and to describe the clinicopathologic features. Sixty patients underwent EUS-guided RTFNA of 64 lesions, including pancreas (n = 45), periluminal lymph nodes (n = 12), mediastinal and retroperitoneal masses (n = 4), and hepatobiliary masses (n = 3). Follow-up data were obtained by surgery, histopathology, or clinical course. Thirty-one lesions were malignant, eight were atypical/suspicious, 16 were non-neoplastic, and nine were non-diagnostic. Of the 55 lesions with sufficient material for interpretation, 54 had follow-up confirmation of the RTFNA diagnosis. The calculated sensitivity and specificity for malignancy was 90% and 100%, respectively. Diagnostic accuracy for malignancy was excellent for gastrointestinal associated lymph nodes (100%), mediastinal and retroperitoneal masses (100%), somewhat less so for pancreatic tumors (94%), and poor for hepatobiliary lesions (33%). EUS-guided RTFNA is accurate for sampling small gastrointestinal tract-associated lesions. EUS-guided RTFNA should be considered as a procedure of choice in selected patients when the results will influence management decisions.
Assuntos
Neoplasias Abdominais/diagnóstico por imagem , Neoplasias Abdominais/patologia , Biópsia por Agulha/métodos , Endossonografia/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Ductos Biliares/diagnóstico por imagem , Ductos Biliares/patologia , Criança , Endoscópios , Endossonografia/instrumentação , Esôfago/diagnóstico por imagem , Esôfago/patologia , Feminino , Humanos , Fígado/diagnóstico por imagem , Fígado/patologia , Linfonodos/diagnóstico por imagem , Linfonodos/patologia , Masculino , Mediastino/diagnóstico por imagem , Mediastino/patologia , Pessoa de Meia-Idade , Pâncreas/diagnóstico por imagem , Pâncreas/patologia , Sensibilidade e EspecificidadeRESUMO
This white paper provides a critical analysis of methods for estimating transporter kinetics and recommendations on proper parameter calculation in various experimental systems. Rational interpretation of transporter-knockout animal findings and application of static and dynamic physiologically based modeling approaches for prediction of human transporter-mediated pharmacokinetics and drug-drug interactions (DDIs) are presented. The objective is to provide appropriate guidance for the use of in vitro, in vivo, and modeling tools in translational transporter science.
Assuntos
Interações Medicamentosas , Proteínas de Membrana Transportadoras/metabolismo , Preparações Farmacêuticas/metabolismo , Farmacocinética , Animais , Disponibilidade Biológica , Transporte Biológico/efeitos dos fármacos , Encéfalo/metabolismo , Guias como Assunto , Humanos , Rim/metabolismo , Fígado/metabolismo , Modelos Biológicos , Distribuição TecidualAssuntos
Comunicação , Relações Enfermeiro-Paciente , Enfermagem Obstétrica , Feminino , Humanos , Gravidez , SemânticaAssuntos
Comunicação Persuasiva , Psicoterapia/métodos , Autoimagem , Privação Sensorial , Adolescente , Adulto , Ansiedade/etiologia , Feminino , Humanos , Masculino , Fatores de TempoRESUMO
BACKGROUND: Activating epidermal growth factor receptor (EGFR) mutations have been implicated in non-small cell lung cancer (NSCLC), and have also been clinically correlated with patient sensitivity to targeted EGFR inhibitors. AIM: To describe a technique for determining EGFR mutation status on archival fine needle aspirate (FNA) specimens from advanced NSCLC patients. METHODS: Eleven archival FNA slides from patients with advanced NSCLC were examined for diagnostic material to identify tumour cell-enriched regions. EGFR mutation status was determined using a slide-scrape DNA extraction protocol of selected tumour cell regions on the smear slides, followed by real time PCR and high resolution melt analysis (HRMAA) of EGFR exons 18, 19, 20, and 21, followed by sequence analysis. RESULTS: All DNA samples were successfully amplified by PCR. Three adenocarcinoma patient samples contained EGFR mutations in exon 19 (L747-P753insS). One of the three had an additional exon 19 mutation (A755D). CONCLUSIONS: Archival cytology slides from patients with NSCLC can be used to determine EGFR mutation status by PCR, HRMAA, and sequencing. The ability to use archival cytology slides greatly increases the potential material available for molecular analysis in diagnosis and selection of patients for targeted therapeutic agents.