RESUMO
BACKGROUND: In recent years, metagenomic Next-Generation Sequencing (mNGS) has increasingly been used for an accurate assumption-free virological diagnosis. However, the systematic workflow evaluation on clinical respiratory samples and implementation of quality controls (QCs) is still lacking. METHODS: A total of 3 QCs were implemented and processed through the whole mNGS workflow: a no-template-control to evaluate contamination issues during the process; an internal and an external QC to check the integrity of the reagents, equipment, the presence of inhibitors, and to allow the validation of results for each sample. The workflow was then evaluated on 37 clinical respiratory samples from patients with acute respiratory infections previously tested for a broad panel of viruses using semi-quantitative real-time PCR assays (28 positive samples including 6 multiple viral infections; 9 negative samples). Selected specimens included nasopharyngeal swabs (n = 20), aspirates (n = 10), or sputums (n = 7). RESULTS: The optimal spiking level of the internal QC was first determined in order to be sufficiently detected without overconsumption of sequencing reads. According to QC validation criteria, mNGS results were validated for 34/37 selected samples. For valid samples, viral genotypes were accurately determined for 36/36 viruses detected with PCR (viral genome coverage ranged from 0.6 to 100%, median = 67.7%). This mNGS workflow allowed the detection of DNA and RNA viruses up to a semi-quantitative PCR Ct value of 36. The six multiple viral infections involving 2 to 4 viruses were also fully characterized. A strong correlation between results of mNGS and real-time PCR was obtained for each type of viral genome (R2 ranged from 0.72 for linear single-stranded (ss) RNA viruses to 0.98 for linear ssDNA viruses). CONCLUSIONS: Although the potential of mNGS technology is very promising, further evaluation studies are urgently needed for its routine clinical use within a reasonable timeframe. The approach described herein is crucial to bring standardization and to ensure the quality of the generated sequences in clinical setting. We provide an easy-to-use single protocol successfully evaluated for the characterization of a broad and representative panel of DNA and RNA respiratory viruses in various types of clinical samples.
Assuntos
Vírus de DNA/genética , Sequenciamento de Nucleotídeos em Larga Escala/normas , Metagenômica/normas , Vírus de RNA/genética , Infecções Respiratórias/virologia , Vírus de DNA/isolamento & purificação , DNA Viral/química , DNA Viral/isolamento & purificação , DNA Viral/metabolismo , Humanos , Controle de Qualidade , Vírus de RNA/isolamento & purificação , RNA Viral/química , RNA Viral/isolamento & purificação , RNA Viral/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Infecções Respiratórias/diagnósticoRESUMO
OBJECTIVE: To compare the diagnostic performance of breast ultrasound and breast magnetic resonance imaging and to correlate the findings on the two techniques that are suggestive of implant rupture. MATERIAL AND METHODS: We reviewed the images and reports of breast ultrasound and magnetic resonance imaging studies done in our diagnostic and interventional breast imaging unit to evaluate breast implants in 205 patients between January 2015 and December 2017. RESULTS: Ultrasound findings were compatible with implant rupture in 87 (42.4%) patients: intracapsular rupture in 44 (21.5%) and intracapsular and extracapsular rupture in 43 (21.0%). Ultrasound yielded 85.2% sensitivity, 89.7% specificity, 86.2% positive predictive value, and 89.0% negative predictive value. Magnetic resonance imaging findings were compatible with implant rupture in 88 (42.9%) patients: intracapsular rupture in 50 (24.4%) and intracapsular and extracapsular rupture in 38 (18.5%). The correlation between positive findings for the location of the rupture on the two imaging techniques was excellent (0.77; p < 0.0001). CONCLUSION: We found high concordance between the two techniques for the detection of intracapsular and extracapsular implant rupture. These results consolidate the use of ultrasound as the first-line imaging technique to evaluate implant integrity in our population; magnetic resonance imaging can be reserved for cases in which the ultrasound diagnosis of implant integrity is uncertain.
Assuntos
Implantes de Mama , Mama , Implantes de Mama/efeitos adversos , Feminino , Humanos , Imageamento por Ressonância Magnética/métodos , Falha de Prótese , Ruptura , Ultrassonografia MamáriaRESUMO
INTRODUCTION/OBJECTIVES: Minimal information exists regarding epicardial pacemaker (EP) implantation in pet ferrets (Mustela putorius furo). The objectives were to describe the indications, surgical technique, and outcome of EP implantation in ferrets for the treatment of advanced atrioventricular block (AVB). ANIMALS, MATERIALS, AND METHODS: Eight client-owned ferrets presenting to five veterinary referral centers. Signalment, physical exam findings, diagnostic tests, anesthesia protocols, surgical implantation techniques, postoperative treatment plans, and EP interrogations were reviewed. Intra- and postoperative, minor and major, and EP-related complications were established. Descriptive statistics were performed to report complication rates. Survival analyses were performed. RESULTS: All ferrets had advanced AVB: 3/8 had high-grade second-degree and 5/8 had third-degree. The primary clinical signs were collapse and weakness. Seven EP were implanted via a transdiaphragmatic approach and one via a left intercostal thoracotomy. Intraoperative complications occurred in 2/8 ferrets, both major. One ferret with severe comorbidities died during general anesthesia. Postoperative pacemaker-related complications were minor: inappropriate sinus beat sensing in 2/8 and occasional muscle fasciculations in 1/8. Two ferrets were alive at the time of manuscript submission, at 10 and 21 months postoperatively. The overall median survival time was 24 months. CONCLUSIONS: Implantation of EP was performed successfully in most ferrets for treatment of advanced AVB and was well tolerated. Ferrets with advanced AVB may experience resolution of clinical signs associated with their cardiac disease following EP implantation. Additional studies are warranted to investigate the effects of epicardial pacing on survival times in this species.
Assuntos
Bloqueio Atrioventricular , Cardiopatias , Marca-Passo Artificial , Animais , Bloqueio Atrioventricular/terapia , Bloqueio Atrioventricular/veterinária , Furões , Cardiopatias/veterinária , Marca-Passo Artificial/efeitos adversos , Marca-Passo Artificial/veterinária , Toracotomia/veterináriaRESUMO
OBJECTIVE: To compare the diagnostic performance of breast ultrasound and breast magnetic resonance imaging and to correlate the findings on the two techniques that are suggestive of implant rupture. MATERIAL AND METHODS: We reviewed the images and reports of breast ultrasound and magnetic resonance imaging studies done in our diagnostic and interventional breast imaging unit to evaluate breast implants in 205 patients between January 2015 and December 2017. RESULTS: Ultrasound findings were compatible with implant rupture in 87 (42.4%) patients: intracapsular rupture in 44 (21.5%) and intracapsular and extracapsular rupture in 43 (21.0%). Ultrasound yielded 85.2% sensitivity, 89.7% specificity, 86.2% positive predictive value, and 89.0% negative predictive value. Magnetic resonance imaging findings were compatible with implant rupture in 88 (42.9%) patients: intracapsular rupture in 50 (24.4%) and intracapsular and extracapsular rupture in 38 (18.5%). The correlation between positive findings for the location of the rupture on the two imaging techniques was excellent (0.77; p<0.0001). CONCLUSION: We found high concordance between the two techniques for the detection of intracapsular and extracapsular implant rupture. These results consolidate the use of ultrasound as the first-line imaging technique to evaluate implant integrity in our population; magnetic resonance imaging can be reserved for cases in which the ultrasound diagnosis of implant integrity is uncertain.
RESUMO
OBJECTIVE: To determine the level of agreement within and between observers in the categorization of breast density on mammograms in a group of professionals using the fifth edition of the American College of Radiology's BI-RADS® Atlas and to analyze the concordance between experts' categorization and automatic categorization by commercial software on digital mammograms. METHODS: Six radiologists categorized breast density on 451 mammograms on two occasions one month apart. We calculated the linear weighted kappa coefficients for inter- and intra-observer agreement for the group of radiologists and between the commercial software and the majority report. We analyzed the results for the four categories of breast density and for dichotomous classification as dense versus not dense. RESULTS: The interobserver agreement among radiologists and the majority report was between moderate and nearly perfect for the analysis by category (κ=0.64 to 0.84) and for the dichotomous classification (κ=0.63 to 0.84). The intraobserver agreement was between substantial and nearly perfect (κ=0.68 to 0.85 for 4 categories and k=0.70 to 0.87 for the dichotomous classification). The agreement between the majority report and the commercial software was moderate both for the four categories (κ=0.43) and for the dichotomous classification (κ=0.51). CONCLUSION: Agreement on breast density within and between radiologists using the criteria established in the fifth edition of the BI-RADS® Atlas was between moderate and nearly perfect. The level of agreement between the specialists and the commercial software was moderate.
Assuntos
Densidade da Mama , Neoplasias da Mama , Mamografia , Neoplasias da Mama/diagnóstico por imagem , Humanos , Mamografia/métodos , Variações Dependentes do Observador , Radiologistas , SoftwareRESUMO
The holistic approach of the human immune system is based on the study of its components collectively driving a functional response to an immunogenic stimulus. To appreciate a specific immune dysfunction, a condition is mimicked ex vivo and the immune response induced is assessed. The application field of such assays are broad and expanding, from the diagnosis of primary and secondary immunodeficiencies, immunotherapy for cancer to the management of patients at-risk for infections and vaccination. These assays are immune monitoring tools that may contribute to a personalised and precision medicine. The purpose of this review is to describe immune functional assays available in the setting of non-HIV acquired immune deficiency. First, we will address the use of theses assays in the diagnosis of opportunistic infections such as viral reactivation. Secondly, we will report the usefulness of these assays to assess vaccine efficacy and to manage immunosuppressive therapies.
Assuntos
Monitoramento de Medicamentos/métodos , Hospedeiro Imunocomprometido , Imunossupressores/uso terapêutico , Infecções Oportunistas/diagnóstico , Humanos , Imunoensaio/métodos , Imunoensaio/normas , Hospedeiro Imunocomprometido/efeitos dos fármacos , Infecções Oportunistas/induzido quimicamente , Infecções Oportunistas/metabolismo , Medicina de Precisão/métodos , Valor Preditivo dos Testes , Fatores de Risco , Ativação Viral/efeitos dos fármacos , Ativação Viral/fisiologia , Viroses/induzido quimicamente , Viroses/diagnósticoRESUMO
The aim of this article is to present an appealing tool for teaching breast imaging that uses the muses of Renaissance and Baroque artists. The examples described here show how medicine and art can be combined to arouse interest in newcomers to breast imaging.
Assuntos
Doenças Mamárias/diagnóstico , Medicina nas Artes , Pinturas , História do Século XVI , História do Século XVII , Humanos , Mamografia , Pinturas/históriaRESUMO
OBJECTIVE: To analyze the imaging characteristics of histologically diagnosed metastases to the breast. MATERIAL AND METHODS: We selected patients histologically diagnosed with metastases to the breast in our diagnostic and interventional breast imaging unit between March 2010 and September 2018. RESULTS: A total of 9 patients (all women; mean age, 60 y; age range, 28-89 y) were diagnosed with metastases to the breast. In 1 (11.11%) case, the primary disease was diagnosed from the breast lesion. The primary tumors were melanoma (n=5), neuroendocrine tumor (n=2, one from the small bowel and one from the cervix), lung adenocarcinoma (n=1), and ovarian cancer (n=1). The clinical and imaging manifestations depend on the type of dissemination of disease and can simulate benign and malignant primary breast lesions. CONCLUSION: There is no specific imaging pattern for metastases to the breast that would help to orient the diagnosis. It is important to consider this etiological possibility if the patient has a history of a primary tumor in another organ.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/secundário , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/patologia , Feminino , Humanos , Pessoa de Meia-Idade , Radiologia , Estudos RetrospectivosAssuntos
Sequência de Aminoácidos/genética , Betacoronavirus/genética , Deleção de Sequência/genética , Proteínas não Estruturais Virais/genética , Sequência de Bases , COVID-19 , Infecções por Coronavirus/epidemiologia , França/epidemiologia , Genoma Viral/genética , Humanos , Pandemias , Pneumonia Viral/epidemiologia , SARS-CoV-2 , Análise de Sequência de RNA , Carga ViralRESUMO
The aim of this study was to determine the HIV subtypes present on Reunion Island, a French island located in the Indian Ocean, where the first case of AIDS was diagnosed in 1987. Paired sera and blood samples were collected between September 1996 and September 1997 from 53 HIV-1-positive patients. Subtyping was performed by serotyping with a previously described subtype-specific enzyme immunoassay (SSEIA) and by genotyping with the heteroduplex mobility assay (HMA). When samples gave uninterpretable results with either of the methods, or discordant results, the V3 env region was sequenced and genetic subtypes were determined by phylogenetic analysis. Genetic subtyping showed that 48 of 53 patients were infected with HIV-1 subtype B (90.5%). This high prevalence of subtype B on Reunion Island is probably due to the regular exchanges with metropolitan France. The other five patients were infected with subtype A (9.5%); they had been directly linked to African populations. Of the 48 subtype B samples, 44 (91.7%) were correctly subtyped by SSEIA and 43 (89.6%) by HMA. However, the SSEIA did not allow the subtyping of A strains in three of five patients. Thus, the SSEIA could be an alternative routine technique for screening subtype B versus nonsubtype B HIV-1 strains.
Assuntos
Soropositividade para HIV/virologia , HIV-1/classificação , Sequência de Aminoácidos , Sequência de Bases , DNA Viral , Feminino , França , Anticorpos Anti-HIV/imunologia , Proteína gp120 do Envelope de HIV/genética , Soropositividade para HIV/sangue , Soropositividade para HIV/imunologia , HIV-1/genética , HIV-1/imunologia , Humanos , Técnicas Imunoenzimáticas , Masculino , Dados de Sequência Molecular , Fragmentos de Peptídeos/genética , Filogenia , Reação em Cadeia da Polimerase/métodos , SorotipagemRESUMO
Skin cancer is the most frequently diagnosed cancer in the United States. Melanoma is the major cause of deaths due to skin cancer, however, squamous cell carcinoma and basal cell carcinoma account for considerable morbidity also. All three types are potentially curable if diagnosed at an early stage. Visual examination of the skin by a trained observer is a simple and effective screening tool which may be utilized in a multitude of settings. Patient education and self-examination augment the impact of office-based screening and mass screening programs.
Assuntos
Programas de Rastreamento , Neoplasias Cutâneas/prevenção & controle , Carcinoma Basocelular/diagnóstico , Carcinoma Basocelular/prevenção & controle , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/prevenção & controle , Humanos , Melanoma/diagnóstico , Melanoma/prevenção & controle , Estadiamento de Neoplasias , Educação de Pacientes como Assunto , Exame Físico , Autoexame , Neoplasias Cutâneas/diagnósticoRESUMO
The relationship was explored between anomalies associated with developmental disabilities and nutritional status of institutionalized children and adolescents who received comprehensive interdisciplinary services. Variables included dietary intake, serum laboratory values, anthropometric measurements, feeding skills assessment, and clinical assessment. Mean intakes of nutrients were greater than 67% of Recommended Daily Allowances. Laboratory values were within normal ranges with the exceptions of serum ferritin, which was borderline or below normal in 46% of the males and 55% of the females, and alkaline phosphatase, which was elevated in 70% of residents receiving anticonvulsants. Energy intake was highest in residents with adequate feeding skills, followed by nonfeeders and feeders requiring behavior management. Subjects had greater triceps skinfold and lower mid-arm muscle circumference and weight measurements as compared to reference data. Findings of this study suggest that these developmentally disabled children and adolescents who received comprehensive interdisciplinary nutritional services, in general, are adequately nourished and have nutrient intakes that meet the RDA requirements.
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Desenvolvimento Infantil , Deficiência Intelectual/fisiopatologia , Estado Nutricional , Atividades Cotidianas , Adolescente , Análise Química do Sangue , Estatura , Peso Corporal , Criança , Ingestão de Energia , Feminino , Humanos , Masculino , Necessidades Nutricionais , Tratamento Domiciliar , Dobras CutâneasAssuntos
Produtos do Gene gag/genética , Variação Genética , Antígenos HIV/genética , Proteína gp120 do Envelope de HIV/genética , Infecções por HIV/virologia , Transcriptase Reversa do HIV/genética , HIV-1/genética , Fragmentos de Peptídeos/genética , Proteínas Virais , Sequência de Aminoácidos , Seguimentos , Infecções por HIV/transmissão , Humanos , Dados de Sequência Molecular , Produtos do Gene gag do Vírus da Imunodeficiência HumanaAssuntos
Complexo AIDS Demência/virologia , Síndrome da Imunodeficiência Adquirida/virologia , DNA Viral/análise , Variação Genética , Proteína gp120 do Envelope de HIV/genética , HIV-1/genética , Fragmentos de Peptídeos/genética , Complexo AIDS Demência/sangue , Complexo AIDS Demência/líquido cefalorraquidiano , Síndrome da Imunodeficiência Adquirida/sangue , Síndrome da Imunodeficiência Adquirida/líquido cefalorraquidiano , Sequência de Aminoácidos , Sequência de Bases , Efeito Citopatogênico Viral , HIV-1/fisiologia , Humanos , Dados de Sequência MolecularAssuntos
Neoplasias Cutâneas/secundário , Adolescente , Neoplasias Encefálicas/patologia , Criança , Pré-Escolar , Doença de Hodgkin/patologia , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Leucemia Mieloide/patologia , Leucemia Mieloide Aguda/patologia , Linfoma Anaplásico de Células Grandes/patologia , Melanoma/secundário , Neuroblastoma/secundário , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Rabdomiossarcoma/secundário , Rabdomiossarcoma Alveolar/secundário , Rabdomiossarcoma Embrionário/secundário , Sarcoma/secundárioAssuntos
Envelhecimento/fisiologia , Envelhecimento da Pele/fisiologia , Envelhecimento/patologia , Ataxia Telangiectasia/patologia , Ataxia Telangiectasia/fisiopatologia , Atrofia , Síndrome de Cockayne/patologia , Síndrome de Cockayne/fisiopatologia , Humanos , Progéria/patologia , Progéria/fisiopatologia , Síndrome de Rothmund-Thomson/patologia , Síndrome de Rothmund-Thomson/fisiopatologia , Pele/patologia , Envelhecimento da Pele/patologia , Síndrome , Síndrome de Werner/patologia , Síndrome de Werner/fisiopatologiaRESUMO
This paper presents an overview of how microsystem technology tools can be applied to the development of rapid, out-of-laboratory measurement capabilities for the determinations of toxigenic fungi and mycotoxins in foodstuffs. Most of the topics discussed are all under investigation within the European Commission-sponsored project Good-Food (FP6-IST). These are DNA arrays, electronic noses and electronic tongues for the detection of fungal contaminants in feed, and biosensors and chemical sensors based on microfabricated electrode systems, antibodies and novel synthetic receptors for the detection of specific mycotoxins. The approach to resolution of these difficult measurement problems in real matrices requires a multidisciplinary approach. The technology tools discussed can provide a route to the rapid, on-site generation of data that can aid the safe production of high-quality foodstuffs.
Assuntos
Análise de Alimentos/métodos , Microbiologia de Alimentos , Fungos/isolamento & purificação , Micotoxinas/análise , Técnicas Biossensoriais , DNA Fúngico/genética , Eletrônica , Fungos/metabolismo , Micotoxinas/biossíntese , Análise de Sequência com Séries de OligonucleotídeosRESUMO
The Kids Count Farm Safety Lesson was delivered to nearly 2,000 fifth-grade students in 15 rural schools in Hillsborough County, Florida. The lesson covered animal, machinery, water, and general safety topics applicable to farming in Florida. A staggered pretest-posttest study design was followed whereby five schools received a multiple-choice pretest and posttest and the remainder of the schools (N = 10) received the posttest only. Results of the study showed a significant increase in the mean number of correct answers on the posttests compared to the pretests. There was no significant difference in the mean number of correct answers of those students who received the pretest and those students who had not, eliminating a "pretest" effect. This study fills an important gap in the literature by evaluating a farm safety curriculum offered in the elementary school setting. It also included migrant schoolchildren in the study population. It is strongly recommended that agricultural safety information be included into the health education curriculum of these elementary schools.
Assuntos
Acidentes de Trabalho/prevenção & controle , Agricultura , Educação em Saúde/normas , Segurança , Criança , Serviços de Saúde da Criança/normas , Currículo , Feminino , Florida , Humanos , Masculino , Desenvolvimento de Programas , Avaliação de Programas e Projetos de Saúde , Serviços de Saúde Escolar/normasRESUMO
We have studied the replication capacity of primary HIV-1 isolates obtained from four AIDS patients in astrocytes. Two patients (P1 and P2) had neurological manifestations without AIDS Dementia Complex (ADC). The other two patients (P3 and P4) had ADC. Two astrocytoma cell lines and normal fetal astrocytes were inoculated with each of these four viral isolates. Viral DNA and mRNA synthesis and also protein accumulation were followed at various times after infection. We found that tumoral as well as fetal astrocytes were susceptible to HIV-1 infection. Three of four viral isolates (P2, P3, P4) were able to infect astrocytes. Both ADC viral isolates (P3, P4) infected astrocytes with identical transcriptional patterns: rev, nef and unspliced mRNAs were expressed for 2 days after infection. The non-ADC patient (P2) with the isolate leading to viral replication in astrocytes had an HIV-1 associated multifocal demyelinating neuropathy. In this case, only nef and unspliced mRNAs were detected a few days after virus inoculation. In all cases, infection of astrocytes was transient and the level of unspliced mRNAs in infected astrocytes was lower than in chronically HIV-1 infected T cells. More extensive work would allow a better understanding of the role of astrocytes in ADC.
Assuntos
Complexo AIDS Demência/virologia , Astrócitos/virologia , Infecções por HIV/virologia , HIV-1/fisiologia , Linhagem Celular , DNA Viral/análise , Embrião de Mamíferos , Produtos do Gene nef/fisiologia , Genes nef , HIV-1/genética , HIV-1/isolamento & purificação , HIV-1/patogenicidade , Humanos , RNA Mensageiro/análise , RNA Viral/análise , Proteínas Virais/análise , Virulência , Replicação Viral , Produtos do Gene nef do Vírus da Imunodeficiência HumanaRESUMO
The purpose of the present study was to assess the viral diversity of hepatitis C virus (HCV) in six nonresponder patients during three unsuccessful treatments. These patients were treated successively with IFN-alpha2a (IFN-alpha) at a posology of 3.10(6) units (MIU) three times a week, 10 MIU three times a week, and a combination of IFN-alpha (3 MIU) plus ribavirin (1,000 mg/day). However, only two chronically infected patients could be included in the study due to the persistence of HCV RNA during the three successive treatments. The viral diversity was analysed by cloning and sequencing the HVR-1 region. The treatment of the two nonresponder patients was associated with the persistence of a wide diversity in the viral population and with the emergence of new or minor variants. Under the influence of standard doses of IFN-alpha, a rearrangement of the quasispecies present was observed at this time point. No significant change in viral load or in the complexity of the quasispecies was observed. A second treatment with a high dose of IFN-alpha induced a significant decrease in the associated viral load and, in one case, resulted in a radical change of the viral diversity. Administration of a combination of IFN-alpha and ribavirin did not affect the evolution of the variants but was followed by the emergence of various multiple variants. These results reinforce the hypothesis of the presence of preexisting quasispecies best adapted to the host environment, and therefore resistant to any current therapy.