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The Belgian law requires that the physician informs his/her patient when he diagnoses a medical reason that makes him/her unfit to drive a vehicle. This paper lists, as they appear in the official texts, the physical conditions and diseases which may be responsible for the inaptitude to drive. Thus are detailed the rules applied in the presence of neurological diseases, psychic disorders, epilepsia, pathological sleepiness, locomotive disorders, cardiovascular disease, rhythm or conduction disturbances, blood pressure abnormalities, coronary or myocardial disease, and hearing loss or vestibular problems. The reader will also find a summary of the rules related to visual ability, alcohol use, driving under the influence of psychotropic drugs, or medicines in general, renal or hepatic diseases, and to the patient who received an organ transplant or an artificial implant. The responsibility of the physician, primarily the general practionner, may be involved in these matters; this issue is discussed.
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Conducción de Automóvil/legislación & jurisprudencia , Medicina Familiar y Comunitaria , Rol del Médico , Bélgica , HumanosRESUMEN
General practitioners often come upon intrafamily abuse cases in their practice. They are either specifically consulted because of these instances of abuse or they discover them by accident. When this happens, they are often unsure of the appropriate procedure to follow, for fear of committing a medical fault or breaking the law. The aim of this short article is not to dwell on the diagnostic methodologies, but to provide a series of general guidelines to follow in everyday practice.
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Violencia Doméstica , Medicina Familiar y Comunitaria , Adulto , Niño , Violencia Doméstica/legislación & jurisprudencia , Medicina Familiar y Comunitaria/normas , Humanos , Registros MédicosAsunto(s)
Receptores de Lipopolisacáridos/metabolismo , Melanoma/fisiopatología , Fagocitos/metabolismo , Neoplasias Cutáneas/fisiopatología , Humanos , Macrófagos/inmunología , Macrófagos/metabolismo , Melanoma/inmunología , Monocitos/inmunología , Monocitos/metabolismo , Fagocitos/inmunología , Neoplasias Cutáneas/inmunologíaRESUMEN
BACKGROUND: MART-1 and gp100 currently are utilized as targets in immunotherapy protocols for metastatic malignant melanoma (MMM). Enrollment of patients into ongoing peptide vaccination trials at the National Cancer Institute includes immunophenotyping of samples of metastatic lesions obtained by fine-needle aspiration (FNA). As therapy progresses, immunocytochemistry is performed on serial FNAs of metastatic lesions to monitor changes in antigen expression during treatment. It is theorized that antigen expression of melanoma cells may be diminished because of selective immunodestruction of tumor cells, or perhaps intentionally, to escape immunosurveillance. METHODS: Thirty-eight lesions from 33 patients were serially monitored for the expression of gp100 (clone HMB-45) and MART-1 (clone M2-7C10), using an avidin-biotin peroxidase technique. The staining intensity of tumor cells was scored on a scale of 0 to 3+, with the proportion of positive cells categorized as less than 25%, 25-50%, 50-75%, and greater than 75%. All lesions were examined within approximately 2 months after the start of peptide vaccination, providing a consistent timepoint for analysis. RESULTS: Using the Wilcoxon signed rank test, the authors found that there were no significant changes from baseline compared with 2 months later for quantitative antigen expression of HMB-45 or MART-1. However, there was a trend toward a decline in staining intensity of tumor cells for HMB-45. CONCLUSIONS: Preliminary results evaluating antigen expression during selective immunotherapy indicate a trend in the decline of staining intensity of tumor cells to HMB-45. Thus, although other studies have shown that peptide-based immunotherapy results in immune selection, this does not hinder the diagnostic utility of antibodies to HMB-45 and MART-1 in FNA samples of MMM.
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Inmunoterapia , Melanoma/inmunología , Glicoproteínas de Membrana/biosíntesis , Proteínas de Neoplasias/biosíntesis , Neoplasias Cutáneas/inmunología , Antígenos de Neoplasias , Biopsia con Aguja , Ensayos Clínicos como Asunto , Diagnóstico Diferencial , Humanos , Inmunohistoquímica , Melanoma/diagnóstico , Melanoma/tratamiento farmacológico , Antígenos Específicos del Melanoma , Metástasis de la Neoplasia , Valor Predictivo de las Pruebas , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/tratamiento farmacológico , Antígeno gp100 del MelanomaRESUMEN
BACKGROUND: Cytology plays a pivotal role in the diagnosis of pleural effusions. In many cases, immunocytochemistry (ICC) is required to elucidate the etiology of the atypical cells. Effusions are samples that present unique problems for ICC. To date there is no standardization of ICC methods for effusions and cytology in general. METHODS: The authors review the most commonly used cytologic preparations, fixatives, and antibodies used in effusion ICC. RESULTS: Through the utilization of cell block preparations and a panel of antibodies appropriate for the differential diagnosis in question, ICC conditions utilized in surgical pathology can be most closely replicated. CONCLUSIONS: ICC may provide reliable insights into various diagnostic dilemmas in effusion cytology, provided that laboratory standardization practices are followed.
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Inmunohistoquímica/normas , Derrame Pleural Maligno/citología , Adenocarcinoma/patología , Anticuerpos Antineoplásicos , Diagnóstico Diferencial , Humanos , Mesotelioma/patologíaRESUMEN
Simian Virus 40 (SV 40) was recently implicated in the pathogenesis of malignant mesothelioma. The oncogenic capacity of SV-40 is a function of a nuclear protein, the large T antigen (Tag). SV-40 Tag DNA sequences are detected by the polymerase chain reaction in 40-80% of malignant mesothelial proliferations. However, the role of immunohistochemistry (IHC) in demonstrating the nuclear localization of Tag is controversial. We sought to determine the clinical utility of SV-40 Tag IHC in pleural effusion cytology as an ancillary tool in the cytologic diagnosis of malignant mesothelioma (MM). Formalin-fixed, paraffin-embedded cell block sections from 100 pleural effusions (32 MMs, 25 benign reactive, 43 metastatic adenocarcinomas) were immunostained for the SV-40 anti-Tag, using two primary monoclonal SV-40 Tag antibodies: clone Pab 416 and clone Pab 101. Despite strong and consistent immunoreactivity in positive controls, no nuclear immunostaining was observed in any case. We believe the small sample size in cytology cell block sections, the low viral copy number in infected cells, and the effect of formalin fixation may have resulted in absence of immunoreactivity. The role of SV-40 Tag IHC in diagnostic cytopathology remains unclear unless further studies reliably show its detection.
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Antígenos Transformadores de Poliomavirus/análisis , Inmunohistoquímica/métodos , Mesotelioma/patología , Virus 40 de los Simios/inmunología , Femenino , Humanos , Masculino , Mesotelioma/química , Mesotelioma/virología , Derrame Pleural Maligno/metabolismo , Derrame Pleural Maligno/patología , Derrame Pleural Maligno/virología , Infecciones por Polyomavirus/metabolismo , Infecciones por Polyomavirus/patología , Infecciones por Polyomavirus/virología , Virus 40 de los Simios/aislamiento & purificación , Infecciones Tumorales por Virus/metabolismo , Infecciones Tumorales por Virus/patología , Infecciones Tumorales por Virus/virologíaRESUMEN
The distinction of mesothelial cells in cytologic samples is often a diagnostic challenge. This is particularly true in potentially malignant effusions in which reactive mesothelial cells may simulate adenocarcinoma (ACA) cells, and in the differentiation of ACA vs. mesothelioma. We sought to determine the superior antibody for the positive identification of mesothelial cells in these circumstances. Cell block sections of 25 reactive and 8 malignant mesothelioma effusions were immunostained with an avidin-biotin procedure, using antibodies to HBME-1 and calretinin. No pretreatment of samples was necessary for the HBME-1-stained slides; microwave antigen retrieval was performed on all slides stained for calretinin. A negative control was performed on each sample. The staining intensity of tumor cells was scored on a scale of 0-3+, with the proportion of immunoreactive cells categorized as <25%, 25-50%, 50-75%, and >75%. The predominant staining pattern for HBME-1 was surface, with rare samples also exhibiting cytoplasmic staining as well. The calretinin-staining pattern was cytoplasmic, with peripheral condensation/prominence and accompanying nuclear staining. All samples were immunoreactive with both antibodies. Fifty-five percent (18/33) of samples showed significantly stronger immunoreactivity with calretinin than with HBME-1; 45% (15/33) of samples showed equivalent staining with the two markers. None of the samples in this study showed stronger immunoreactivity with HBME-1 than with calretinin. Sixty-one percent (20/33) of samples stained with HBME-1 at a moderate (2+) intensity. Fifty-five percent (18/33) of samples stained with calretinin at a strong (3+) intensity. While only 12% of samples showed >75% immunoreactivity for HBME-1, 58% of samples showed >75% of cells immunoreactive for calretinin. Calretinin is the preferred marker in identifying mesothelial cells in cytologic samples, showing the highest sensitivity for mesothelial cells, as evidenced by a more intense staining reaction in a higher percentage of cells than with HBME-1. Published 2001 Wiley-Liss, Inc.
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Antígenos de Neoplasias/inmunología , Biomarcadores de Tumor/inmunología , Células Epiteliales/patología , Inmunoglobulina G , Derrame Pleural/diagnóstico , Proteína G de Unión al Calcio S100/inmunología , Calbindina 2 , Recuento de Células , Citodiagnóstico/métodos , Diagnóstico Diferencial , Humanos , Técnicas para InmunoenzimasRESUMEN
The melanoma patient's immune response to tumor has been extensively studied. Yet, the frequently observed coexistence of tumor-associated Ag (TAA)-specific T cells with their target cells in vivo remains unexplained. Loss of TAA expression might contribute to this paradox. We studied TAA expression in metastases by obtaining fine-needle aspirations from 52 tumor lesions in 30 patients with melanoma before and soon after immunotherapy. Limitations due to low amounts of starting material were overcome with a high fidelity antisense RNA amplification method. TAA expression was measured by quantitative real-time PCR of anti-sense RNA. Decrease in gp100/Pmel-17 TAA preceded tumor disappearance in several instances and could be best explained by immune selection because most patients had received gp100/Pmel-17-specific vaccination. Conversely, immune selection was absent in nonregressing lesions. These observations suggest that vaccination, when successful, triggers a broad inflammatory reaction that can lead to tumor destruction despite immune selection. Additionally, lack of clinical response might be attributed to lack of this initiating event rather than immune escape. This study provides an insight into the natural history of tumors and defines a strategy for the characterization of gene expression in tumors during therapy.
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Antígenos de Neoplasias/biosíntesis , Vacunas contra el Cáncer/inmunología , Proteínas de Neoplasias/biosíntesis , Adulto , Anciano , Antígenos de Neoplasias/genética , Vacunas contra el Cáncer/administración & dosificación , Femenino , Amplificación de Genes , Regulación Neoplásica de la Expresión Génica/inmunología , Humanos , Cinética , Antígeno MART-1 , Masculino , Melanoma/genética , Melanoma/inmunología , Melanoma/secundario , Persona de Mediana Edad , Proteínas de Neoplasias/genética , Regresión Neoplásica Espontánea , ARN sin Sentido/genética , Testículo/inmunología , Factores de Tiempo , Células Tumorales CultivadasAsunto(s)
Anticuerpos Antineoplásicos/inmunología , Antígenos de Neoplasias/inmunología , Melanoma/inmunología , Proteínas de Neoplasias/inmunología , Neoplasias de las Glándulas Suprarrenales/inmunología , Neoplasias de las Glándulas Suprarrenales/patología , Especificidad de Anticuerpos , Biopsia con Aguja , Neoplasias Óseas/inmunología , Neoplasias Óseas/patología , Humanos , Neoplasias Hepáticas/inmunología , Neoplasias Hepáticas/patología , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/patología , Antígeno MART-1 , Melanoma/patología , Metástasis de la Neoplasia , Neoplasias Pancreáticas/inmunología , Neoplasias Pancreáticas/patologíaRESUMEN
Loss of heterozygosity (LOH) at the 3p region is found in up to 50% of epithelial ovarian neoplasms. The von Hippel-Lindau (VHL) gene at the 3p25 locus is one of the tumor-suppressor genes located at 3p. The role, if any, of the VHL gene locus is not clear in ovarian carcinogenesis. We analyzed primary and metastatic ovarian clear-cell carcinomas (OCCC) for LOH at 3p25 to determine its frequency and its diagnostic utility as an adjunctive tool in the differential diagnosis of metastatic clear-cell carcinomas. Microdissection followed by single-step DNA extraction and polymerase chain reaction (PCR) amplification, using two polymorphic markers flanking the VHL gene locus, was done on archival histology and cytology samples from 9 patients with metastatic OCCC. Of the informative cases, 43% of the metastatic and 50% of the primary OCCC showed LOH. LOH at the VHL gene locus is not uncommon in clear-cell ovarian carcinoma. LOH at 3p25 in cytologic specimens may be a valuable adjunct in the diagnosis of OCCC metastasis in cytologically equivocal cases. OCCC should enter the differential in clear-cell carcinomas of unknown primary that show LOH at 3p25. Published 2001 Wiley-Liss, Inc.
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Adenocarcinoma de Células Claras/genética , Adenocarcinoma de Células Claras/secundario , Cromosomas Humanos Par 3/genética , Disección , Ligasas , Pérdida de Heterocigocidad/genética , Neoplasias Ováricas/genética , Neoplasias Ováricas/secundario , Reacción en Cadena de la Polimerasa , Proteínas Supresoras de Tumor , Ubiquitina-Proteína Ligasas , Adenocarcinoma de Células Claras/patología , Biomarcadores de Tumor , Diagnóstico Diferencial , Femenino , Genes Supresores de Tumor/genética , Humanos , Neoplasias Ováricas/patología , Proteínas/genética , Estudios Retrospectivos , Proteína Supresora de Tumores del Síndrome de Von Hippel-LindauAsunto(s)
Biomarcadores/análisis , Derrame Pleural Maligno/química , Proteína G de Unión al Calcio S100/análisis , Adenocarcinoma/química , Adenocarcinoma/secundario , Neoplasias de la Mama/química , Neoplasias de la Mama/secundario , Calbindina 2 , Núcleo Celular/química , Núcleo Celular/patología , Neoplasias del Colon/química , Neoplasias del Colon/patología , Citodiagnóstico , Citoplasma/química , Citoplasma/patología , Humanos , Técnicas para Inmunoenzimas , Neoplasias Pulmonares/química , Neoplasias Pulmonares/patología , Mesotelioma/química , Mesotelioma/secundario , Derrame Pleural Maligno/patologíaRESUMEN
Assessment of antigen expression by solid tumors has relied predominantly on immunohistochemistry, flow cytometry, and more recently quantitative real-time polymerase chain reaction. However, all these techniques present intrinsic limits. The laser scanning cytometer, by combining the properties of light and fluorescence microscopy with those of laser cytometry, can quantitatively and objectively analyze hypocellular samples such as fine-needle aspirates on an individual cell basis. To validate the fidelity of laser scanning cytometry for quantitative immunophenotyping of fine-needle aspirates, the authors measured the expression of the melanoma-associated antigens MART-1 and gp100 as well as HLA-A2, a HLA class 1 restriction element associated with their recognition by melanoma-specific T cells. Expression of melanoma antigens and HLA was measured by laser scanning cytometry and immunohistochemistry in fine-needle aspirates from melanoma metastases. In addition, transcription levels of both melanoma antigens were recorded by quantitative real-time polymerase chain reaction. A quantity of less than 1,000 cells per sample (average 682 cells) was sufficient for the analysis. Laser scanning cytometry estimates correlated with those of immunohistochemistry and quantitative real-time polymerase chain reaction for MART-1 and gp100. A good correlation in HLA-A2 detection by laser scanning cytometry and immunohistochemistry was also observed. Moreover, the laser scanning cytometer could discriminate subsets of cells from the same lesion with heterogeneous melanoma antigen expression, leading to the observation that cells with a DNA index greater than 2.5 expressed significantly less gp100. Thus, laser scanning cytometry yields detailed information on protein expression in individual cells and represents a new tool for dissecting the immune response in the tumor microenvironment.
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Antígenos de Neoplasias/biosíntesis , Antígeno HLA-A2/biosíntesis , Melanoma/inmunología , Glicoproteínas de Membrana/biosíntesis , Proteínas de Neoplasias/biosíntesis , Antígenos de Neoplasias/genética , Biopsia con Aguja , Citometría de Flujo/métodos , Antígeno HLA-A2/genética , Humanos , Citometría de Imagen/métodos , Rayos Láser , Antígeno MART-1 , Melanoma/patología , Glicoproteínas de Membrana/genética , Metástasis de la Neoplasia , Proteínas de Neoplasias/genética , Células Tumorales Cultivadas , Antígeno gp100 del MelanomaRESUMEN
BACKGROUND: Tyrosinase, the rate-limiting enzyme in melanin synthesis, is a melanoma associated antigen that is recognized by both CD4+ and CD8+ T-cells in an HLA-restricted fashion. Peptides derived from the tyrosinase antigen currently are being utilized as a target for T-cells in several immunotherapy protocols for metastatic malignant melanoma (MMM) at the National Institutes of Health/National Cancer Institute. Serial fine-needle aspirations of metastatic lesions are performed to monitor the antigen expression of tyrosinase during treatment by immunostaining cytologic preparations with the monoclonal antibody T311. METHODS: In the current study, 62 samples of MMM were evaluated for tyrosinase immunoreactivity on air-dried, acetone fixed cytospins and the corresponding formalin fixed, paraffin embedded cell block using an avidin-biotin immunoperoxidase method. RESULTS: Positive immunoreactivity revealed a granular cytoplasmic staining in melanocytic cells. The current study results showed that 92% of samples (57 of 62) were T311 immunoreactive on cell block preparations, whereas only 61% (38 of 62) were immunoreactive on cytospin preparations. In 66% of samples (41 of 62) immunoreactivity for T311 was greater in the cell block sample than in the corresponding cytospin, whereas in only 3% of samples (2 of 62) was it greater in the cytospins. In 31% of samples (19 of 62) there was no significant difference in immunoreactivity between the 2 sample types. CONCLUSIONS: The results of the current study show that tyrosinase is a sensitive marker for the detection of MMM; however, the optimal method of sample preparation for immunoperoxidase staining appears to be formalin fixation and paraffin embedding as tyrosinase immunoreactivity is diminished significantly in air-dried cytospin samples despite subsequent acetone fixation. Cancer (Cancer Cytopathol)
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Melanoma/enzimología , Monofenol Monooxigenasa/inmunología , Acetona , Anticuerpos Monoclonales , Biopsia con Aguja , Centrifugación/métodos , Fijadores , Formaldehído , Humanos , Técnicas para Inmunoenzimas , Oxígeno/química , Adhesión en Parafina , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Fijación del Tejido/métodosRESUMEN
Evaluation for circulating tumor cells and bone marrow micrometastases has generated considerable interest due to a potential association with disease recurrence and poor prognosis. In this study, we examined bone marrow and apheresis samples from Stage II, III, and IV patients (n 120) enrolled in various clinical breast cancer trials at the National Institutes of Health/National Cancer Institute. For each patient sample, two Diff-Quik-stained cytospins were reviewed for morphology, and approximately 1 x 10(6) cells were analyzed for the expression of cytokeratins using an avidin-biotin immunoperoxidase method. Keratin-positive malignant cells appearing as single cells or in small clusters were detected in bone marrow samples from Stage IV patients only (9/68, 13%) and detected in apheresis samples from both Stage III and IV patients (13/245, 5%). These findings indicate that the combination of cytomorphology with immunocytochemistry can be utilized for the investigation of circulating tumor cells and bone marrow micrometastases, and that positive results appear to correlate with high tumor stage/burden.
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Biomarcadores de Tumor , Neoplasias de la Mama/patología , Queratinas , Femenino , Humanos , Inmunohistoquímica , Metástasis de la Neoplasia/diagnóstico , Metástasis de la Neoplasia/patología , Estadificación de NeoplasiasRESUMEN
Mechanisms of drug resistance other than P-glycoprotein are of increasing interest as the list of newly identified members of the ABC transport family has grown. We sought to characterize the phenotype of the newly discovered ABC transporter encoded by the mitoxantrone resistance gene, MXR, also known as ABCP1 or BCRP. The pharmacodynamics of mitoxantrone and 12 other fluorescent drugs were evaluated by confocal microscopy in four multidrug-resistant human colon (S1) and breast (MCF-7) cancer cell lines. We utilized two sublines, MCF-7 AdVp3000 and S1-M1-80, and detected overexpression of MXR by PCR, immunoblot assay and immunohistochemistry. These MXR overexpressing sublines were compared to cell lines with P-glycoprotein- and MRP-mediated resistance. High levels of cross-resistance were observed for mitoxantrone, the anthracyclines, bisantrene and topotecan. Reduced levels of mitoxantrone, daunorubicin, bisantrene, topotecan, rhodamine 123 and prazosin were observed in the two sublines with high MXR expression. Neither the P-glycoprotein substrates vinblastine, paclitaxel, verapamil and calcein-AM, nor the MRP substrate calcein, were extruded from MCF-7 AdVp3000 and S1-M1-80 cells. Thus, the multidrug-resistant phenotype due to MXR expression is overlapping with, but distinct from, that due to P-glycoprotein. Further, cells that overexpress the MXR protein seem to be more resistant to mitoxantrone and topotecan than cells with P-glycoprotein-mediated multidrug resistance. Our studies suggest that the ABC half-transporter, MXR, is a potent, new mechanism for conferring multiple drug resistance. Definition of its mechanism of transport and its role in clinical oncology is required.
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Transportadoras de Casetes de Unión a ATP/genética , Neoplasias de la Mama/genética , Neoplasias del Colon/genética , Resistencia a Múltiples Medicamentos , Resistencia a Antineoplásicos , Regulación Neoplásica de la Expresión Génica/genética , Proteínas de Transporte de Membrana , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Antineoplásicos/farmacocinética , Northern Blotting , Proteínas de Unión al ADN/genética , Colorantes Fluorescentes/farmacocinética , Humanos , Inmunohistoquímica , Microscopía Confocal , Mitoxantrona/farmacocinética , Proteína 2 Asociada a Resistencia a Múltiples Medicamentos , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/genética , Proteína 3 Homóloga de MutS , Reacción en Cadena de la Polimerasa , ARN Mensajero/análisis , Proteínas Ribosómicas/genética , Células Tumorales CultivadasRESUMEN
An ongoing multicenter U.S. clinical study evaluates the safety and effectiveness of creating linear lesions with radiofrequency (RF) energy using a 3.7 Fr microcatheter used with a 9 Fr steerable guiding catheter in the right atrium (RA) to treat paroxysmal atrial fibrillation (PAF). Study entry criteria were symptomatic, drug refractory PAF. RF energy was delivered in 60-second episodes at 30 W or 50 W power settings. Electrode tissue contact was ascertained by sharp electrograms of high amplitude. Coagulum Index (C.I.), a new calculation to predict the probability of coagulum development, was derived analytically from measurements of RF power, RF current, and time to reach maximum temperature in each ablation episode. The RF data from 398 separate ablation episodes in 15 patients were used to calculate C.I. Coagulum presence was determined by postablation visual inspection of the microcatheter electrodes. A logistic model was used to estimate the probability of coagulum occurrence, with C.I. as the predictive variable. When C.I. > or = 12, the probability of coagulum formation increased significantly in this model (P < 0.0001). Prolonging the power delivery rise time and limiting maximum RF power setting to 30 W effectively lowers C.I. and minimizes coagulum formation. These results should also apply to left atrium ablation, where minimizing coagulum formation may decrease the risk of stroke or mortality.
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Fibrilación Atrial/cirugía , Ablación por Catéter/efectos adversos , Modelos Logísticos , Trombosis/etiología , Trombosis/prevención & control , Ablación por Catéter/métodos , Suministros de Energía Eléctrica/normas , Humanos , Medición de Riesgo , Temperatura , Estados UnidosRESUMEN
The standard of practice in cytopathology does not include an individual specimen triage (IST) for sample optimization, but rather prescribes a uniform procedure, e.g., for smears, cell blocks, and cytospins. IST requires additional resources. We sought to evaluate whether IST would result in enhanced diagnostic accuracy and specimen turnaround time in effusions. In order to evaluate the efficacy of IST, 50 effusion samples (31 pleural, 16 peritoneal, and 3 pericardial), each with a minimum volume of 50 ml, were utilized. Each sample was prepared via IST to include at least two initial prepared Diff-Quik-stained cytospins on which the IST was based, as well as a standard cytopreparation protocol for nontriaged samples (NTS) which was limited to 3 smears (2 Papanicolaou-stained, and 1 Diff-Quik-stained) and a hematoxylin-eosin (H&E)-stained cell block section. All triaged and NTS were reviewed retrospectively to determine if IST offered any advantages over the standard cytopreparation protocol for effusion samples. Each was evaluated for diagnostic concordance, turnaround time for final diagnosis, and optimal preparation. In 46 cases, diagnoses in IST and NTS were 100% concordant. Four cases showed minor discrepancies between the original and the NTS diagnoses. In general, the discordant cases were due to sparse cellularity in a specimen composed largely of blood. There was no difference in turnaround time for final diagnosis. Based on a review of all samples, the combination of cell block preparation and cytospins (stained with Diff-Quik and Papanicolaou stains) were considered optimal for microscopic evaluation. IST offers no practical advantage over the NTS standard specimen preparation in relation to the accuracy of final diagnosis or turnaround time. The lysing of grossly bloody fluids with subsequent preparation of cytospins yielded superior preparations for microscopic evaluation over NTS. The standard preparation of effusion samples should include the preparation of a cell block, and cytospins stained with Diff-Quik and Papanicolaou stains, for optimal microscopic evaluation.
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Líquido Ascítico/patología , Derrame Pericárdico/patología , Derrame Pleural/patología , Adulto , Estudios de Evaluación como Asunto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Estudios Prospectivos , Manejo de EspecímenesRESUMEN
The establishment of melanoma cell lines from fine-needle aspiration biopsies (FNAB) has allowed for an enhanced understanding of the complex interactions that occur between T cells and tumor cells. The technique of FNAB offers the advantage of providing a sequential analysis of the same tumor nodules throughout treatment. The expression of melanoma antigens (MAs) was assessed in fresh melanoma FNAB samples and from tumor cell lines derived from these samples using several different approaches. Cytospin preparations of freshly isolated tumor cell explants were analyzed by immunocytochemistry (ICC), while the daughter cell line was analyzed by fluorescent activated cell sorting (FACS) analysis, and semiquantitative and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR, qRT-PCR). As assessed by these methods, the level of MA expression by the original tumor cell explants correlated with the expression in established in vitro cell lines. Molecular analysis of the established cell lines utilizing PCR technology improved the sensitivity of detection of MA expression. Thus FNAB of melanoma is an efficient and effective method of tissue procurement, capable of generating, sequentially and from the same lesion, fresh tumor cells, tumor infiltrating lymphocytes (TIL), and long-term melanoma cell lines.
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Biopsia con Aguja/métodos , Técnicas de Cultivo de Célula/métodos , Melanoma/metabolismo , Células Tumorales Cultivadas , Citocinas/metabolismo , Citometría de Flujo , Prueba de Histocompatibilidad , Humanos , Inmunohistoquímica , Melanoma/patología , Metástasis de la Neoplasia , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
BACKGROUND: Melanoma antigen (MA)-specific vaccination strongly enhances antitumor reactivity in vivo and is capable of producing strong cytotoxic T lymphocyte responses in vitro. Furthermore, specific human leukocyte antigen-restricted T cell activation is hypothesized to occur in response to peptide-based immunotherapy, which may lead to the preferential killing of tumor cells bearing the relevant MA. The development of melanoma antigen-loss variants may subsequently occur in vivo. METHODS: Analysis of 532 melanoma lesions from 204 patients was performed on fine-needle aspiration biopsy specimens. Lesions were graded for the expression of the MAs gp100 and MART-1 with use of immunocytochemistry. A total of 351 melanoma lesions were divided into cohorts on the basis of the treatment received. The pretreatment group (n = 175) consisted of lesions obtained before any form of gp100 immunotherapy, with the posttreatment group (n = 176) consisting of lesions obtained after vaccination with a modified gp100 epitope, gp209-2M +/- interleukin 2 (IL-2). RESULTS: The percentage of lesions not expressing the gp100 antigen is greater than the percentage not expressing MART-1 (26% vs 14%). The frequency of lesions with high expression (> 75%) of gp100 significantly decreased with therapy (47% vs 34%) and conversely negative lesions increased (18% vs 29%). Treatment of lesions with peptide alone (no IL-2) revealed a significant decrease in gp100 expression (47% vs 32%), enhanced with the addition of IL-2 to therapy (47% vs 35%). The expression of MART-1 remained essentially unchanged unless IL-2 was added (54% vs 54%, MART-1 peptide alone, 54% vs 43%, MART-1 peptide + IL-2). Of 94 patients (181 lesions) assessed for gp100 expression before treatment, 10 patients responded to therapy. Pretreatment lesions in responding patients expressed some level of gp100 in all cases compared with 27% of nonresponding lesions, which were negative for gp100 expression. CONCLUSIONS. This study indirectly demonstrates that vaccination with an MA-derived peptide can result in immune selection in vivo. Furthermore, it provides strong immunologic evidence for the specificity of MA vaccines and to the relevance of MA expression in predicting the response to vaccination.