Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 24
Filtrar
1.
Children (Basel) ; 10(1)2022 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-36670568

RESUMO

Deep neck space infections (DNSI) are defined as infections in the potential spaces and fascial planes of the neck. We show the clinical case of a retro and para-pharyngeal abscess in a healthy 5-year-old child complicated by compression and dislocation of the larynx with marked airway caliber reduction and potentially fatal extension up to the mediastinal aditus. DNSI can occur at any age and, due to its rapid progression, requires immediate treatment in children. In healthy children, concurrent abscesses in separate neck spaces are rare. DNSI recurrence should alert the physician to the possibility of a congenital problem, and if imaging fails, laryngoscopy may be the best diagnostic technique.

2.
Int J Mol Sci ; 18(7)2017 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-28696386

RESUMO

Human Papillomavirus (HPV) E6/E7 mRNA test demonstrated high specificity in detecting HPV infections, but studies assessing its efficacy in terms of cancer risk stratification are lacking. Follow-up studies are arduous and expensive. Biobank would be the answer to the problem, although data investigating the effects of long-term storage on RNA preservation are still needed. We addressed these issues by retrieving 202 residual liquid-based cervical specimens, collected from 149 women attending cervical cancer screening during the years 2001-2012. Samples were stored in Adriatic Biobank at room temperature and without any handing. After calculation of RNA yield and purity, E6/E7 mRNA test was retrospectively performed on each samples, to assess analytic and diagnostic performances. Using automated extraction procedures, RNA of good quantity and quality was obtained. The mean value of RNA concentration was 27.5 ng/µL. The mean A260/A280 ratio was 2.1. An invalid mRNA test result was found in 11.9% of the specimens. Neither RNA integrity, nor analytic performances of mRNA test were influenced by the year of sample collection. In total, 62.4% of the specimens tested as mRNA positive; among these, 89.2% were CIN2+. E6/E7 mRNA was detected in all Squamous Cervical Cancer (SCC) cases. Percentage of positive samples increased with the severity of histological diagnosis. mRNA testing, showing specificity and predictive values of 75.6% and 84.4%, respectively, significantly improved the corresponding values for DNA testing. Thus, the reflex mRNA test was demonstrated to be suitable to triage women with persistent cervical lesions. A "one sample for all" approach is possible, with practical benefits for Biobank-based long-term longitudinal studies, diseases prevention, prediction, diagnosis and treatment.


Assuntos
Colo do Útero/metabolismo , Proteínas Oncogênicas Virais/genética , Papillomaviridae/genética , Proteínas E7 de Papillomavirus/genética , RNA Mensageiro/genética , RNA Viral/genética , Bancos de Espécimes Biológicos , Feminino , Humanos , Estudos Longitudinais , Medicina de Precisão/métodos , RNA Mensageiro/análise , Estudos Retrospectivos
3.
Acta Cytol ; 59(4): 345-54, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26343271

RESUMO

OBJECTIVE: This study was designed to evaluate whether the adjunct of human telomerase RNA component (hTERC) fluorescence in situ hybridization (FISH) analysis to cytological diagnosis and human papillomavirus (HPV)-DNA testing may serve as a predictive marker for distinguishing cervical lesions destined to regress from those at high risk of progression towards invasive cancer. STUDY DESIGN: hTERC FISH analysis was performed on 54 residual liquid-based cytology specimens obtained from women referred to colposcopy for the detection of atypical squamous cells of undetermined significance or worse (ASCUS+) lesions. Histological diagnosis was considered the gold standard and cervical intraepithelial neoplasia of grade 2 or worse (CIN2+) as the worst outcome. RESULTS: Oncogenic HPV-DNA was found in 96.3% of the specimens. Among these, 38.5% revealed a CIN2+ diagnosis. hTERC gene amplification was detected in 37% of the cases; among these, 70% showed up as CIN2+. hTERC FISH analysis significantly improves the specificity and positive predictive value of HPV-DNA testing, thus differentiating patients with a CIN2+ diagnosis from those with a CIN2- diagnosis. CONCLUSIONS: Despite the limitation of a small study sample, our findings provide promising data, indicating the possible role of hTERC analysis in the assessment of the risk of developing cervical cancer. This approach would implement the specificity of DNA testing, avoiding overtreatment at the same time. Prospective follow-up studies are needed with the aim of introducing hTERC FISH into decision-making algorithms.


Assuntos
DNA Viral/genética , RNA/genética , Telomerase/genética , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Adulto , Idoso , Colposcopia/métodos , Feminino , Amplificação de Genes , Testes de DNA para Papilomavírus Humano/métodos , Humanos , Hibridização in Situ Fluorescente/métodos , Pessoa de Meia-Idade , Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/patologia , Infecções por Papillomavirus/virologia , Neoplasias do Colo do Útero/virologia , Adulto Jovem , Displasia do Colo do Útero/diagnóstico , Displasia do Colo do Útero/patologia , Displasia do Colo do Útero/virologia
5.
Expert Rev Mol Diagn ; 15(1): 137-50, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25245757

RESUMO

OBJECTIVE: Colposcopy is widely used to triage women with mild cervical abnormalities. However, this approach is associated with low specificity and predictive value. The efficacy of E6/E7 mRNA test for this purpose has been demonstrated, but studies estimating its cost-effectiveness are still lacking. Given the limited healthcare financial resources, such an evaluation is a priority. METHODS: We analyzed the clinical history of 432 women referred to colposcopy and colposcopy-directed biopsy for persisting ASCUS and LSIL, and compared three alternative triage protocols: immediate colposcopy; reflex HPV DNA testing and HPV DNA plus mRNA tests in sequence. RESULTS: Molecular tests in sequence significantly reduce colposcopy referral, cost for assessed women, and cost for CIN2 detected. On the other hand, incremental cost-effectiveness ratio of this protocol was the highest. CONCLUSION: Our preliminary data, providing an estimation of the economic burden deriving from the introduction of E6/E7 mRNA test in the triage algorithm of patients with mild cervical abnormalities, may be useful for future healthcare policy.


Assuntos
Células Escamosas Atípicas do Colo do Útero , Detecção Precoce de Câncer/economia , Infecções por Papillomavirus/diagnóstico , Lesões Intraepiteliais Escamosas Cervicais/diagnóstico , Neoplasias do Colo do Útero/diagnóstico , Adulto , Colposcopia , Análise Custo-Benefício , Feminino , Humanos , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular/economia , Papillomaviridae/genética , Proteínas E7 de Papillomavirus/genética , RNA Mensageiro/genética , Lesões Intraepiteliais Escamosas Cervicais/virologia , Neoplasias do Colo do Útero/virologia
6.
Acta Cytol ; 58(5): 495-500, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25358681

RESUMO

OBJECTIVES: Endometrial cytology offers a reliable alternative to biopsy in endometrial cancer detection and it may be useful in obtaining material to study prognostic and predictive markers. Over the years, new sampling devices have been developed. Molecular alterations in endometrial cancers were previously described using formalin-fixed paraffin-embedded tissues with particular attention, in endometrioid carcinomas, to the PTEN-PI3K pathway. PTEN evaluation could be useful in endometrial carcinomas for selecting patients for target therapies. STUDY DESIGN: We studied 51 endometrial samples collected using the Endogyn device and 71 obtained with the Endoflower dispositive device, and processed using liquid-based cytology. Most of the cases were matched with a corresponding histological biopsy. The overall accuracy of Endoflower was 100%. Immunohistochemistry (IHC) and immunocytochemistry (ICC) for PTEN were performed using monoclonal antibody 6H2.1 from DAKO. RESULTS: The IHC showed PTEN-null glands in 4 cases. The same cancers were negative in ICC. Among the 10 carcinomas on cytology, PTEN-null glands were found in 1 case. All the normal endometrium control cases were positive in cytology and histology. CONCLUSIONS: Our results suggest that endometrial devices provide useful material for the diagnosis and evaluation of PTEN expression.


Assuntos
Biomarcadores Tumorais/genética , Carcinoma Endometrioide/diagnóstico , Neoplasias do Endométrio/diagnóstico , PTEN Fosfo-Hidrolase/genética , Manejo de Espécimes/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais , Biópsia , Carcinoma Endometrioide/genética , Carcinoma Endometrioide/patologia , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/patologia , Endométrio , Feminino , Expressão Gênica , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Manejo de Espécimes/instrumentação
7.
Regul Pept ; 190-191: 18-24, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24809812

RESUMO

AIM OF THE STUDY: The gastrointestinal peptide hormone ghrelin (Ghr) was discovered in 1999 as the endogenous ligand for the growth hormone secretagogue receptor (GHSR-1a). It is a pleiotropic peptide that modulates a wide spectrum of biological activities, such as growth hormone (GH) release, feeding stimulation, adiposity and cardiovascular actions. The presence of Ghr mRNA in the iris and ciliary body (CB) epithelium was recently demonstrated in animal models, where a possible myorelaxing effect on the iris muscles has been suggested. Based on these observations, the aim of our study was to investigate the Ghr and GHSR-1a expression and localization in the normal human eye. MATERIAL: Five different ciliary body/iris samples from normal eyes were subjected to Western blot analysis. Immunohistochemical detection was performed on three enucleated eyes. Twenty aqueous humor (AqH) samples obtained from patients submitted to cataract surgery were analyzed with an ELISA for the presence of Ghr. RESULTS: Ghr and GHSR-1a were co-expressed by the pigmented epithelium (PE) of the CB, by the retinal pigmented epithelium (RPE) and by the anterior limiting layer (ALL) of the iris. No reaction was detected at the subepithelial level in the ciliary or pupillae smooth muscle cells. The AqH samples were positive for the presence of Ghr. CONCLUSION: This study provides the first evidence that Ghr and GHSR-1a are expressed in the human eye by specific cells. The understanding of the functional role of Ghr at the human eye level needs more efforts and investigation, but a hypothetical action on the GH retinal synthesis and/or on the circadian clock system could be suggested.


Assuntos
Olho/metabolismo , Grelina/análise , Receptores de Grelina/análise , Grelina/biossíntese , Humanos , Receptores de Grelina/biossíntese
8.
Clin Interv Aging ; 9: 683-8, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24790420

RESUMO

The introduction of organized cervical cancer (CC) screening programs has drastically reduced the prevalence of CC. However the incidence is still too high, especially among elderly women. All guidelines strongly recommend a regular Papanicolaou (Pap) testing for young and middle-aged patients. On the other hand, many international professional societies no longer advise screening in women who have undergone hysterectomy, and in women aged 65 years and above, who have a previous history of regular Pap smears. Here we report the case of poorly differentiated CC, involving the pelvic lymph nodes and urinary bladder, occurring in a 79-year-old woman who regularly underwent Pap tests, with no reported cytological abnormalities. In this very rare case, the CC cells, as well as cells from metastatic lymph nodes and cells from urinary specimens, molecularly showed human papilloma virus (HPV)-53. With the limitations of a single case, this report brings important information to prevent CC in elderly patients: the utility of molecular tests to increase sensitivity of Pap smears in postmenopausal women; the importance of HPV-53 as one of the four "emergent" genotypes having a possible role in oncogenesis; and the presence of HPV-53 in lymph node metastases from cervical carcinoma, which would support the role of this virus in the maintenance of malignant status.


Assuntos
Carcinoma de Células Escamosas/etiologia , Teste de Papanicolaou , Papillomaviridae , Infecções por Papillomavirus/complicações , Neoplasias do Colo do Útero/etiologia , Idoso , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/virologia , Colo do Útero/patologia , Reações Falso-Negativas , Feminino , Humanos , Papillomaviridae/genética , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase em Tempo Real , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia
9.
PLoS One ; 9(2): e88769, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24586389

RESUMO

Metal ions are necessary for the proper functioning of the immune system, and, therefore, they might have a significant influence on the interaction between bacteria and host. Ionic dyshomeostasis has been recently observed also in cystic fibrosis (CF) patients, whose respiratory tract is frequently colonized by Stenotrophomonas maltophilia. For the first time, here we used an inductively mass spectrometry method to perform a spatial and temporal analysis of the pattern of changes in a broad range of major trace elements in response to pulmonary infection by S. maltophilia. To this, DBA/2 mouse lungs were comparatively infected by a CF strain and by an environmental one. Our results showed that pulmonary ionomic profile was significantly affected during infection. Infected mice showed increased lung levels of Mg, P, S, K, Zn, Se, and Rb. To the contrary, Mn, Fe, Co, and Cu levels resulted significantly decreased. Changes of element concentrations were correlated with pulmonary bacterial load and markers of inflammation, and occurred mostly on day 3 post-exposure, when severity of infection culminated. Interestingly, CF strain - significantly more virulent than the environmental one in our murine model - provoked a more significant impact in perturbing pulmonary metal homeostasis. Particularly, exposure to CF strain exclusively increased P and K levels, while decreased Fe and Mn ones. Overall, our data clearly indicate that S. maltophilia modulates pulmonary metal balance in a concerted and virulence-dependent manner highlighting the potential role of the element dyshomeostasis during the progression of S. maltophilia infection, probably exacerbating the harmful effects of the loss of CF transmembrane conductance regulator function. Further investigations are required to understand the biological significance of these alterations and to confirm they are specifically caused by S. maltophilia.


Assuntos
Fibrose Cística/metabolismo , Pneumopatias/metabolismo , Pneumopatias/microbiologia , Stenotrophomonas maltophilia/patogenicidade , Oligoelementos/metabolismo , Virulência/fisiologia , Animais , Cobalto/metabolismo , Cobre/metabolismo , Ferro/metabolismo , Magnésio/metabolismo , Masculino , Manganês/metabolismo , Camundongos , Fósforo/metabolismo , Rutênio/metabolismo , Zinco/metabolismo
11.
Biomed Res Int ; 2013: 453606, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24369532

RESUMO

Although HPV-DNA test and E6/E7 mRNA analyses remain the current standard for the confirmation of human papillomavirus (HPV) infections in cytological specimens, no universally adopted techniques exist for the detection of HPV in formalin-fixed paraffin-embedded samples. Particularly, in routine laboratories, molecular assays are still time-consuming and would require a high level of expertise. In this study, we investigated the possible use of a novel HPV tyramide-based chromogenic in situ hybridization (CISH) technology to locate HPV on tissue specimens. Then, we evaluate the potential usefulness of p16(INK4a)/Ki-67 double stain on histological samples, to identify cervical cells expressing HPV E6/E7 oncogenes. In our series, CISH showed a clear signal in 95.2% of the specimens and reached a sensitivity of 86.5%. CISH positivity always matched with HPV-DNA positivity, while 100% of cases with punctated signal joined with cervical intraepithelial neoplasia grade 2 or worse (CIN2+). p16/Ki67 immunohistochemistry gave an interpretable result in 100% of the cases. The use of dual stain significantly increased the agreement between pathologists, which reached 100%. Concordance between dual stain and E6/E7 mRNA test was 89%. In our series, both CISH and p16(INK4a)/Ki67 dual stain demonstrated high grade of performances. In particular, CISH would help to distinguish episomal from integrated HPV, in order to allow conclusions regarding the prognosis of the lesion, while p16(INK4a)/Ki67 dual stain approach would confer a high level of standardization to the diagnostic procedure.


Assuntos
Inibidor p16 de Quinase Dependente de Ciclina/isolamento & purificação , Antígeno Ki-67/genética , Proteínas Oncogênicas Virais/isolamento & purificação , Neoplasias do Colo do Útero/genética , Inibidor p16 de Quinase Dependente de Ciclina/genética , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Humanos , Imuno-Histoquímica , Hibridização In Situ , Antígeno Ki-67/isolamento & purificação , Proteínas Oncogênicas Virais/genética , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/genética , Infecções por Papillomavirus/patologia , Infecções por Papillomavirus/virologia , Inclusão em Parafina , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia
12.
PLoS One ; 8(2): e57600, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23460880

RESUMO

Pap test, and especially HPV DNA test, identify a large group of women who do not have any clinically relevant lesions, i.e., CIN2+ (Cervical Intraepithelial Neoplasia grade 2 or worse), but who are at greater risk of getting lesions in the future. The follow up of these women needs new biomarkers with prognostic value. The objective of this study is to evaluate the prognostic value of E6/E7 mRNA over-expression assay (PreTect HPV-Proofer, Norchip) for 5 HR-HPV types (16, 18, 31, 33, and 45) for progression to CIN2+ after a negative colposcopy. This prospective study, conducted at four Italian centres, enrolled 673 women with either a negative colposcopy or a negative or CIN1 histology. The clinical end-point was histological confirmation of CIN2+. Women were classified at baseline according to mRNA results and managed according to local colposcopy protocols. At least one conclusive follow-up test was obtained for 347 women (25 months average lapse since recruitment, range 5-74). Only seven CIN2+ were detected during follow up, three among the 82 women positive for mRNA at baseline, two among the 250 negative (Fisher exact test, p = 0.02), and two among the 12 with an invalid test. Absolute CIN2+ risk was 6.7/1,000 person/years in the whole cohort. The absolute CIN2+ risk was 18.4/1,000 person/years and 3.6/1,000 person/years in mRNA-positive and mRNA-negative women, respectively. In conclusion, E6/E7 mRNA over-expression appears to be a good candidate as a prognostic biomarker to manage HR-HPV DNA-positive women with negative colposcopy or histology, particularly in order to decrease follow-up intensity in those who are negative.


Assuntos
Colposcopia , Detecção Precoce de Câncer/métodos , Proteínas Oncogênicas Virais/genética , Papillomaviridae/genética , Displasia do Colo do Útero/patologia , Displasia do Colo do Útero/virologia , Adolescente , Adulto , Idoso , Feminino , Seguimentos , Regulação Viral da Expressão Gênica , Humanos , Pessoa de Meia-Idade , Proteínas Oncogênicas Virais/metabolismo , Prognóstico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Adulto Jovem
13.
J Med Virol ; 85(3): 472-82, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23280876

RESUMO

High-risk human papillomavirus (HR-HPV) genotype viral load and E6/E7 mRNA detection are proposed as surrogate markers of malignant cervical lesion progression. Currently, the use of commercially available DNA-based or mRNA-based tests is under investigation. In this study, the viral DNA load and E6/E7 mRNA detection of the five most common HR-HPV types detected in cervical cancer worldwide were compared in 308 cervical samples by using in-house type-specific quantitative real-time PCR assays and PreTect HPV-Proofer test, respectively. Sensitivity and negative predictive values were higher for the HPV-DNA assays combined (95.0% and 96.0%, respectively) than the RNA assays (77.0% and 88.0%, respectively); conversely, the mRNA test showed a higher specificity and higher positive predictive value (81.7% and 66.9%, respectively) than the DNA test (58.6% and 52.5%, respectively) for detecting histology-confirmed high-grade cervical intraepithelial neoplasia. A significantly higher association between viral DNA load and severity of disease was observed for HPV 16 and 31 (γ = 0.62 and γ = 0.40, respectively) than for the other HPV types screened. A good degree of association between the two assays was found for detection of HPV 16 (k = 0.83), HPV 18 (k = 0.72), HPV 33 (k = 0.66), and HPV 45 (k = 0.60) but not for HPV 31 (k = 0.24). Sequence analysis in L1 and E6-LCR regions of HPV 31 genotypes showed a high level of intra-type variation. HR-HPV viral DNA load was significantly higher in E6/E7 mRNA positive than negative samples (P < 0.001), except for HPV 31. These findings suggest that transcriptional and replicative activities can coexist within the same sample.


Assuntos
DNA Viral/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Papillomaviridae/isolamento & purificação , RNA Mensageiro/isolamento & purificação , RNA Viral/isolamento & purificação , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/virologia , Adulto , Idoso , DNA Viral/genética , Feminino , Genótipo , Humanos , Pessoa de Meia-Idade , Papillomaviridae/genética , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/virologia , Valor Preditivo dos Testes , RNA Mensageiro/genética , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Estudos Retrospectivos , Sensibilidade e Especificidade , Análise de Sequência de DNA , Adulto Jovem
14.
Gynecol Oncol ; 128(3): 427-32, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23200910

RESUMO

OBJECTIVE: This two-arm longitudinal study was performed within a regional organized cervical-cancer-prevention program in which HPV-DNA test is used in primary screening. The aim was to analyze the diagnostic performances of p16INK4a/Ki-67 dual-test and E6/E7-mRNA test in identifying CIN2+ lesion among HPV-DNA positive (HPV-DNAve) women triaged for LSIL-or-worse liquid based cytology (LBC). METHODS: Thirty-six thousand thirty-one women participated to HPV-DNA screening program pilot study. Three thousand six hundred forty-one resulted HPV-DNAve; among these, 43% were LSIL-or-worse (LSIL+). HPV-DNAve/LSIL+ patients were submitted to colposcopy and histological assessment of any visible lesions. Dual-test was performed on 794 residual LBC specimens. In 405 cases, dual-test result was related to histology, considering CIN2+ as endpoint. mRNA test has been carried out retrospectively, on a subset of 173 residual LBC specimens. RESULTS: Agreement between dual-test and histological diagnosis was 59%. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of cytology-plus-dual-test approach were 62.3%, 76.8%, 63.1% and 84.2%, respectively. Dual-test improved specificity, PPV and NPV of cytological triage Agreement between mRNA testing and histology was 65%. Cytology-plus mRNA testing showing sensitivity, specificity, PPV and NPV reaching 32.1%, 94.9%, 75% and 50%, respectively; implemented specificity and PPV of cytology alone in triaging DNA-ve/LSIL+ patients (p<0.01). CONCLUSIONS: We provided promising data indicating the important role that p16(INK4)/Ki-67 dual-test, and mostly E6/E7 mRNA test, might have in triaging HPV-DNAve. These approaches would exclude the occurrence of cervical cancer and would avoid overtreatment, at the same time. Further longitudinal analysis has to be considered.


Assuntos
DNA Viral/análise , Papillomaviridae/genética , Infecções por Papillomavirus/prevenção & controle , Infecções por Papillomavirus/virologia , Neoplasias do Colo do Útero/prevenção & controle , Neoplasias do Colo do Útero/virologia , Adulto , Inibidor p16 de Quinase Dependente de Ciclina , DNA Viral/genética , Detecção Precoce de Câncer/métodos , Feminino , Humanos , Estudos Longitudinais , Pessoa de Meia-Idade , Proteínas de Neoplasias , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/epidemiologia , RNA Mensageiro/análise , RNA Mensageiro/genética , Estudos Retrospectivos , Resultado do Tratamento , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/epidemiologia , Proteínas do Envelope Viral/genética
15.
Clin Cancer Res ; 19(3): 691-8, 2013 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-23243218

RESUMO

PURPOSE: The therapeutic choice for patients with lung adenocarcinoma depends on the presence of EGF receptor (EGFR) mutations. In many cases, only cytologic samples are available for molecular diagnosis. Bronchoalveolar lavage (BAL) and pleural fluid, which represent a considerable proportion of cytologic specimens, cannot always be used for molecular testing because of low rate of tumor cells. EXPERIMENTAL DESIGN: We tested the feasibility of EGFR mutation analysis on BAL and pleural fluid samples by next-generation sequencing (NGS), an innovative and extremely sensitive platform. The study was devised to extend the EGFR test to those patients who could not get it due to the paucity of biologic material. A series of 830 lung cytology specimens was used to select 48 samples (BAL and pleural fluid) from patients with EGFR mutations in resected tumors. These samples included 36 cases with 0.3% to 9% of neoplastic cells (series A) and 12 cases without evidence of tumor (series B). All samples were analyzed by Sanger sequencing and NGS on 454 Roche platform. A mean of 21,130 ± 2,370 sequences per sample were obtained by NGS. RESULTS: In series A, EGFR mutations were detected in 16% of cases by Sanger sequencing and in 81% of cases by NGS. Seventy-seven percent of cases found to be negative by Sanger sequencing showed mutations by NGS. In series B, all samples were negative for EGFR mutation by Sanger sequencing whereas 42% of them were positive by NGS. CONCLUSIONS: The very sensitive EGFR-NGS assay may open up to the possibility of specific treatments for patients otherwise doomed to re-biopsies or nontargeted therapies.


Assuntos
Adenocarcinoma/genética , Líquido da Lavagem Broncoalveolar/química , Receptores ErbB/genética , Neoplasias Pulmonares/genética , Mutação , Derrame Pleural Maligno/química , Adenocarcinoma de Pulmão , Análise Mutacional de DNA , Éxons , Sequenciamento de Nucleotídeos em Larga Escala , Humanos
16.
Curr Pharm Des ; 19(8): 1450-7, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23016778

RESUMO

BACKGROUND: Loop Electrosurgical Excision Procedure (LEEP) represents the mainstay technique for CIN2+ removal. The major concern in conservative treatment is to verify whether CIN eradication was complete, since incomplete excision is associated with an increased risk of cervical cancer. The histopathologic evaluation of resection margins status is far from perfect, since cervical lesions may recur in 5-15% of patients who had conisation specimens with clean margins. Current follow-up protocol of patients treated by conisation for high grade CIN is manly based on the combination of cytology-plus- HPV-DNA testing. This approach showed high sensitivity but low specificity level in detecting recurrence. The consequence were overdiagnosis and overtreatment, especially in youngest women, in which spontaneous regression rate of CIN is substantial. In this longitudinal study we investigated whether patient's age, cone depth and pre-conisation HPV-load level, may be used as predictive markers for residual/recurrent CIN after conisation. Then we aimed to examined the role of E6/E7 mRNA testing during post-conization follow-up. METHODS: The study, focused on the outcome of 116 patients treated for CIN by LEEP, included three consecutive steps. Firstly, the authors analysed the prevalence of residual/recurrence disease after conization; then, they investigated which factors may influence treatment failure even when resection margins were clean; finally, they evaluated the diagnostic accuracy of E6/E7 mRNA test as predictive marker of recurrence. RESULTS: HPV infection was detected in 31% of patients at 6-month follow-up and in 11.2% of patients, at 24-month follow-up. Younger women showed higher rate of recurrence than older ones. The risk of residual/recurrent infection did not correlate with cone-depth. Recurrence is higher in patients with low viral load level than in those having high load levels. mRNA test showed higher specificity and positive predictive value than the combination cytology-plus-HPV-DNA test. CONCLUSION: The inclusion of mRNA test within the current protocol of follow-up would efficiently and earlier predict the risk of residual/ recurrent cervical abnormalities after conisation. This molecular strategy would also reduce overtreatment, particularly in patients above 30 years of age.


Assuntos
Alphapapillomavirus/isolamento & purificação , Neoplasias do Colo do Útero/diagnóstico , Adulto , Alphapapillomavirus/genética , Feminino , Seguimentos , Humanos , Recidiva , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologia
17.
Urol Int ; 88(2): 232-4, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22248535

RESUMO

Dystrophic calcified nodule of the testicle represents an exceptional lesion with an unknown etiology and controversial diagnostic approach. There are very few reports in the literature that have examined this feature. Here, we report the fourth case of dystrophic testis nodule, identified in a 46-year-old man who presented the urologist with a palpable mass on his left testicle and no other symptoms. Histopathological findings were discussed and pooled with those of all previously published series. We believe that this description is particularly innovative because of the absence of symptoms and testicular pain associated with the lesion, and interesting as an attempt to determine the recognition of this rare entity.


Assuntos
Calcinose/diagnóstico , Doenças Testiculares/diagnóstico , Neoplasias Testiculares/diagnóstico , Testículo/patologia , Biópsia , Calcinose/patologia , Calcinose/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade , Palpação , Valor Preditivo dos Testes , Terminologia como Assunto , Doenças Testiculares/classificação , Doenças Testiculares/patologia , Doenças Testiculares/cirurgia , Neoplasias Testiculares/classificação , Neoplasias Testiculares/patologia , Testículo/diagnóstico por imagem , Testículo/cirurgia , Ultrassonografia Doppler em Cores
18.
Ann Clin Lab Sci ; 41(4): 353-9, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22166505

RESUMO

Immunofluorescence is the most frequently utilized technique to analyze protein expression. Fixed immunofluorescent cell suspensions, however, can only be stored for a week. We investigated whether liquid-based cytology could be used to detect antigens in cultured cells after a long storage period. Murine and human cells were fixed in PreservCyt solution, stored for various periods, and then used to perform an automated immunocytochemical analysis. Phosphorylation of the nuclear transcription factor Stat-5 induced by IL-7 was detected up to 4 months after IL-7 stimulation. Simultaneous nuclear positivity for the proliferation index MIB-1 and membrane positivity for the CD30 antigen were evident three months after fixation. Liquid-based cytology thus ensures long-lasting nuclear and membrane antigen immunoreactivity and permits the storage of cells from laborious experiments at room temperature for future analyses.


Assuntos
Antígenos/metabolismo , Membrana Celular/metabolismo , Núcleo Celular/metabolismo , Técnicas Citológicas/métodos , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/metabolismo , Preservação Biológica , Animais , Linhagem Celular Tumoral , Membrana Celular/efeitos dos fármacos , Núcleo Celular/efeitos dos fármacos , Humanos , Interleucina-7/farmacologia , Antígeno Ki-1/metabolismo , Antígeno Ki-67/metabolismo , Leucócitos Mononucleares/efeitos dos fármacos , Camundongos , Fosforilação/efeitos dos fármacos , Fator de Transcrição STAT5/metabolismo , Soluções , Coloração e Rotulagem , Fatores de Tempo
19.
J Clin Microbiol ; 49(7): 2643-50, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21525231

RESUMO

There is evidence that testing for human papillomavirus (HPV) E6/E7 mRNA is more specific than testing for HPV DNA. A retrospective study was carried out to evaluate the performance of the PreTect HPV-Proofer E6/E7 mRNA assay (Norchip) as a triage test for cytology and HPV DNA testing. This study analyzed 1,201 women, 688 of whom had a colposcopy follow-up and 195 of whom had histology-confirmed high-grade intraepithelial neoplasia or worse (CIN2+). The proportion of positive results and the sensitivity and specificity for CIN2+ were determined for HPV mRNA in comparison to HPV DNA and cytology. All data were adjusted for follow-up completeness. Stratified by cytological grades, the HPV mRNA sensitivity was 83% (95% confidence interval [CI] = 63 to 94%) in ASC-US (atypical squamous cells of undetermined significance), 62% (95% CI = 47 to 75%) in L-SIL (low-grade squamous intraepithelial lesion), and 67% (95% CI = 57 to 76%) in H-SIL (high-grade squamous intraepithelial lesion). The corresponding figures were 99, 91, and 96%, respectively, for HPV DNA. The specificities were 82, 76, and 45%, respectively, for HPV mRNA and 29, 13, and 4%, respectively, for HPV DNA. Used as a triage test for ASC-US and L-SIL, mRNA reduced colposcopies by 79% (95% CI = 74 to 83%) and 69% (95% CI = 65 to 74%), respectively, while HPV DNA reduced colposcopies by 38% (95% CI = 32 to 44%) and by 15% (95% CI = 12 to 19%), respectively. As a HPV DNA positivity triage test, mRNA reduced colposcopies by 63% (95% CI = 60 to 66%), having 68% sensitivity (95% CI = 61 to 75%), whereas cytology at the ASC-US+ threshold reduced colposcopies by 23% (95% CI = 20 to 26%), showing 92% sensitivity (95% CI = 87 to 95%). In conclusion, PreTect HPV-Proofer mRNA can serve as a better triage test than HPV DNA to reduce colposcopy referral in both ASC-US and L-SIL. It is also more efficient than cytology for the triage of HPV DNA-positive women. Nevertheless, its low sensitivity demands a strict follow-up of HPV DNA positive-mRNA negative cases.


Assuntos
DNA Viral/isolamento & purificação , Detecção Precoce de Câncer/métodos , Proteínas Oncogênicas Virais/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , RNA Mensageiro/isolamento & purificação , Neoplasias do Colo do Útero/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Técnicas Citológicas/métodos , DNA Viral/genética , Feminino , Humanos , Pessoa de Meia-Idade , Papillomaviridae/genética , Infecções por Papillomavirus/virologia , RNA Mensageiro/genética , Estudos Retrospectivos , Sensibilidade e Especificidade , Estados Unidos , Adulto Jovem
20.
J Cytol ; 27(1): 35-6, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21042534

RESUMO

Cutaneous leishmania life cycle images on cytology smears are very rare. We report herein a gallery of cytologic images from a case of sporadic cutaneous leishmaniasis in a 61 year old man presenting with ulcerative skin lesion.

SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...