Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 2.381
Filtrar
1.
Clín. investig. ginecol. obstet. (Ed. impr.) ; 46(3): 107-114, jul.-sept. 2019. tab
Artigo em Espanhol | IBECS | ID: ibc-182716

RESUMO

Objetivo: Estudio prospectivo de 4 años (2015-2018) de casos de ASCUS y LSIL en la citología cervicovaginal y su control citológico evolutivo en 3 meses. Diseño: Estudio de: edad, paridad, abortos, nuligestas, menopáusicas, métodos contraceptivos, sexualidad y disfunciones sexuales, antecedentes patológicos; citología y virus del papiloma humano, comparando la evolución en ambos grupos. Resultados: La serie son 146 casos de ASCUS y 82 casos de LSIL. Hubo diferencias significativas en ASCUS, mayor en edad (p<0,02), paridad (p<0,001) y abortos (p<0,02). Hubo diferencia significativa en LSIL, mayor en nuligestas (p<0,001). Hubo diferencia significativa, p<0,02, en uso de condón, mayor en LSIL. En ambos grupos, >50% de los casos, no utilizaban ningún método anticonceptivo. No hubo diferencias significativas en la sexualidad ni en las disfunciones sexuales, ni en los antecedentes, entre ambos grupos. ASCUS se asoció a infecciones en el 15,05% y LSIL se asoció a infecciones en el 9,74%, sin diferencias. Hubo diferencia significativa en virus del papiloma humano negativo, p<0,01, mayor en ASCUS, 43,15% versus 20,73% en LSIL. Hubo diferencia significativa en otros virus de alto riesgo positivos, p<0,01, mayor en LSIL. En el control citológico a 3 meses hubo diferencia significativa en ASCUS+vaginosis bacteriana, p<0,02, mayor en LSIL. También en el grupo LSIL hubo más LSIL en el control a 3 meses en el 26,82% (p<0,01). En el seguimiento a un año no hubo diferencias significativas entre ambos grupos. Hubo citología negativa respectivamente en el 64,86% y en el 47,82%. Conclusión: La repetición de la citología a 3 meses tiene valor para descartar los falsos positivos y detectar las infecciones asociadas para poder tratarlas, lo que redundaría en anticipar la normalidad citológica


Objective: Prospective study of 4 years (2015-2018) of the ASCUS and LSIL cases in the vaginocervical cytology, and your evolutive cytological control in 3 months. Design: Study of: age, parity, nulligravides, menopausal, contraceptive methods, sexuality and sexual dysfunctions, pathologic antecedents, cytology and HPV (human papillomavirus) analysis, comparing the evolution in both groups. Results: The series are 146 cases of ASCUS and 82 cases of LSIL. In ASCUS, there were significant differences in age (P<.02), parity (P<.001), and abortions (P<.02). In LSIL, there were significant differences in nulligravides (P<.001). There were significant differences (P<.02) in condom use in LSIL. In both groups more of 50% of the cases dońt use any contraceptive method There were no significant differences in sexuality nor in sexual dysfunctions, neither in antecedents. ASCUS was associated with infections in 15,05%. LSIL was associated with infections in 9,74%, without differences. There were significant differences in negative HPV (P<.001) in ASCUS, 43,15% versus 20,73% in LSIL. There were significant differences in High Risk other viruses positive (P<.001) in LSIL. In the cytologic control at 3 monts, there were significant differences in ASCUS+BV (bacterial vaginosis) (P<.02) in LSIL. Also in LSIL group there were more LSIL in the cytologic control at 3 months, 26,82% (P<.01). In the follow-up at 1 year, there were no significant differences. The negative cytology was in 64,86% and 47,82%, respectively. Conclusion: The reiterative cytology in 3 months are value for detecting the associated infections, and treated; to run over and advance the cytologic normality


Assuntos
Humanos , Feminino , Gravidez , Adulto , Pessoa de Meia-Idade , Idoso , Vagina/citologia , Lesões Intraepiteliais Escamosas Cervicais/patologia , Células Escamosas Atípicas do Colo do Útero/citologia , Neoplasia Intraepitelial Cervical/patologia , Técnicas Citológicas/métodos , Esfregaço Vaginal/métodos , Estudos Prospectivos
2.
Nat Protoc ; 14(8): 2546-2570, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31341291

RESUMO

Distal cholesterol biosynthesis (CB) has recently taken center stage as a promising drug target in several diseases previously not linked to this biochemical pathway, including cardiovascular disease, cancer, multiple sclerosis and Alzheimer's disease. Most enzymes involved in this pathway are hard to isolate, warranting dedicated analytical tools for biochemical screening. We describe the use of gas chromatography-electron ionization mass spectrometry (GC-MS) in a whole-cell screening assay aimed at monitoring interactions with all enzymes of distal CB in a single experiment. Following cell culture and lipid extraction, the trimethylsilyl ethers of sterols are analyzed by GC-MS. Analytical data for 23 relevant sterols (intermediates) are provided, allowing their unambiguous identification. Sterol pattern analysis reveals the target enzyme on the basis of characteristic marker sterols, whereas quantification of 2-13C-acetate incorporation correlates with the inhibitory activity of drug candidates. The protocol can be used by both experienced scientists and newcomers to the field, allowing detection and quantification of small molecule-enzyme interactions in distal CB. The entire protocol can be carried out within two working days.


Assuntos
Colesterol/metabolismo , Técnicas Citológicas/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Bioensaio/métodos , Colesterol/análise , Colesterol/química , Sistemas de Liberação de Medicamentos , Células HL-60 , Humanos , Esteróis/análise , Esteróis/química , Esteróis/metabolismo
3.
Biotechnol Lett ; 41(8-9): 929-939, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31321593

RESUMO

OBJECTIVE: To develop a simple robust methodology of screening multiple CHO cell clones secreting recombinant proteins to assess their specific productivity. RESULTS: We developed a dual assay based on immunoassay measurements of a recombinant protein expression combined with staining of viable cells with resazurin. Following this approach, colonies can be simultaneously assessed for cell growth rate and for production of a recombinant protein. Combination of these two assays enables to estimate productivity of a recombinant protein per cell from the very early stages of a cell line development process (CLD) and exclude poor producers from further steps. Comparison of the dual assay with a standard CLD protocol followed by only analysis of protein expression level showed at least 10-20% increase in the amount of clones that can be included into pool of high-producers at early stages. This shortens duration of a typical CLD scheme from 23 to 19 weeks. CONCLUSIONS: Our method: (i) allows to include into workflow clones that demonstrate slow growth during single cell cloning but producing high amounts of a target protein, which otherwise would be lost in standard protocols of cells screening; (ii) can be applied for testing of DNA vectors for transfection and protein production; (iii) can be used for monitoring the heterogeneity of cell population and analysis of stable pools productivity.


Assuntos
Biotecnologia/métodos , Células CHO , Proliferação de Células , Técnicas Citológicas/métodos , Programas de Rastreamento/métodos , Proteínas Recombinantes/metabolismo , Animais , Cricetulus , Proteínas Recombinantes/genética , Coloração e Rotulagem/métodos
4.
Appl Spectrosc ; 73(11): 1292-1298, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31219327

RESUMO

The development of new methods for the detection of redox cycle is important for biological and clinical diagnoses. Here, a new cyclometalated iridium complex, (4-(2-pyridyl) benzaldehyde)2Ir (5-chloro-1,10-phenanthroline) ([(4-pba)2Ir(5-Cl-phen)]PF6, probe 1), has been synthesized and applied to rapid, sensitive, and reversible detection and imaging of redox cycle HSO3-/H2O2 in living cells. The probe 1 is synthesized by using 4-(2-pyridyl) benzaldehyde as main ligand and 5-chloro-1,10-phenanthroline as ancillary ligand. Probe 1 exhibited "off-on-off" photoluminescence (PL) signal change in response to HSO3- and H2O2 in aqueous solution within 1 min. The change of PL intensity is proportional to HSO3- concentration from 40 µM to 300 µM and to H2O2 concentration from 40 µM to 260 µM. The detection limit is 10 µM for HSO3- and 20 µM for H2O2. Additionally, probe 1 was applied to detect HSO3- in food samples with satisfactory results. More importantly, PL imaging of HeLa cells indicates that probe 1 is able to image redox cycle HSO3-/H2O2 in living cells.


Assuntos
Complexos de Coordenação/química , Corantes Fluorescentes/química , Peróxido de Hidrogênio/análise , Irídio/química , Sulfitos/análise , Técnicas Citológicas/métodos , Células HeLa , Humanos , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/metabolismo , Oxirredução , Espectrometria de Fluorescência/métodos , Sulfitos/química , Sulfitos/metabolismo
5.
PLoS Biol ; 17(5): e3000279, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31100061

RESUMO

We report the development and characterization of a method, named reversible association with motor proteins (RAMP), for manipulation of organelle positioning within the cytoplasm. RAMP consists of coexpressing in cultured cells (i) an organellar protein fused to the streptavidin-binding peptide (SBP) and (ii) motor, neck, and coiled-coil domains from a plus-end-directed or minus-end-directed kinesin fused to streptavidin. The SBP-streptavidin interaction drives accumulation of organelles at the plus or minus end of microtubules, respectively. Importantly, competition of the streptavidin-SBP interaction by the addition of biotin to the culture medium rapidly dissociates the motor construct from the organelle, allowing restoration of normal patterns of organelle transport and distribution. A distinctive feature of this method is that organelles initially accumulate at either end of the microtubule network in the initial state and are subsequently released from this accumulation, allowing analyses of the movement of a synchronized population of organelles by endogenous motors.


Assuntos
Técnicas Citológicas/métodos , Proteínas Motores Moleculares/metabolismo , Organelas/metabolismo , Estreptavidina/metabolismo , Axônios/metabolismo , Axônios/ultraestrutura , Transporte Biológico , Biotina/metabolismo , Dendritos/metabolismo , Dendritos/ultraestrutura , Células HeLa , Humanos , Organelas/ultraestrutura , Reprodutibilidade dos Testes
6.
Adv Colloid Interface Sci ; 269: 309-333, 2019 Jul.
Artigo em Espanhol | MEDLINE | ID: mdl-31128462

RESUMO

Cell-cell and cell-matrix adhesions are fundamental in all multicellular organisms. They play a key role in cellular growth, differentiation, pattern formation and migration. Cell-cell adhesion is substantial in the immune response, pathogen-host interactions, and tumor development. The success of tissue engineering and stem cell implantations strongly depends on the fine control of live cell adhesion on the surface of natural or biomimetic scaffolds. Therefore, the quantitative and precise measurement of the adhesion strength of living cells is critical, not only in basic research but in modern technologies, too. Several techniques have been developed or are under development to quantify cell adhesion. All of them have their pros and cons, which has to be carefully considered before the experiments and interpretation of the recorded data. Current review provides a guide to choose the appropriate technique to answer a specific biological question or to complete a biomedical test by measuring cell adhesion.


Assuntos
Adesão Celular/fisiologia , Técnicas Citológicas/métodos , Pinças Ópticas , Animais , Técnicas Citológicas/instrumentação , Transferência Ressonante de Energia de Fluorescência/métodos , Humanos , Microscopia de Força Atômica
7.
Vet Pathol ; 56(5): 725-731, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31113293

RESUMO

Digital microscopy (DM) has been employed for primary diagnosis in human medicine and for research and teaching applications in veterinary medicine, but there are few veterinary DM validation studies. Region of interest (ROI) digital cytology is a subset of DM that uses image-stitching software to create a low-magnification image of a slide, then selected ROI at higher magnification, and stitches the images into a relatively small file of the embedded magnifications. This study evaluated the concordance of ROI-DM compared to traditional light microscopy (LM) between 2 blinded clinical pathologists. Sixty canine and feline cytology samples from a variety of anatomic sites, including 31 cases of malignant neoplasia, 15 cases of hyperplastic or benign neoplastic lesions, and 14 infectious/inflammatory lesions, were evaluated. Two separate nonblinded adjudicating clinical pathologists evaluated the reports and diagnoses and scored each paired case as fully concordant, partially concordant, or discordant. The average overall concordance (full and partial concordance) for both pathologists was 92%. Full concordance was significantly higher for malignant lesions than benign. For the 40 neoplastic lesions, ROI-DM and LM agreed on general category of tumor type in 78 of 80 cases (98%). ROI-DM cytology showed robust concordance with the current gold standard of LM cytology and is potentially a viable alternative to current LM cytology techniques.


Assuntos
Doenças do Gato/patologia , Técnicas Citológicas/métodos , Doenças do Cão/patologia , Processamento de Imagem Assistida por Computador , Microscopia/métodos , Animais , Doenças do Gato/diagnóstico por imagem , Gatos , Doenças Transmissíveis/diagnóstico por imagem , Doenças Transmissíveis/patologia , Doenças Transmissíveis/veterinária , Doenças do Cão/diagnóstico por imagem , Cães , Inflamação/diagnóstico por imagem , Inflamação/patologia , Inflamação/veterinária , Neoplasias/diagnóstico por imagem , Neoplasias/patologia , Neoplasias/veterinária , Software
8.
BMC Cancer ; 19(1): 296, 2019 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-30940100

RESUMO

BACKGROUND: High-grade serous ovarian cancer is a detrimental disease. Treatment options in patients with a recurrent disease are dependent on BRCA1/2 mutation status since only patients with known BRCA mutation are eligible for treatment with poly(ADP-ribose) polymerase inhibitors (PARPi). The aim of this study was to compare concordance of BRCA mutation analyses from cytological samples (CS) with BRCA mutation analyses from histological formalin fixed paraffin embedded (FFPE) samples. METHODS: Mutation analysis of BRCA1 and BRCA2 genes was performed in 44 women diagnosed with primary or recurrent high-grade ovarian cancer from three different samples: blood, cytological sample (ascites, pleural effusion and enlarged lymph nodes) and tumor tissue. Results from all three samples were compared. RESULTS: Among 44 patients, there were 15 germline mutations and two somatic mutations. A 100% concordance was found between cytological and histologic samples. CONCLUSION: There is a 100% concordance in BRCA mutation testing between cytological and histologic samples. BRCA mutation testing from CS could replace testing from FFPE tissue in clinical decision making in ovarian cancer patients. TRIAL REGISTRATION: The study was retrospectively registered at ISRCTN registry on 24/11/2015 - ISRCTN42408038 .


Assuntos
Proteína BRCA1/genética , Proteína BRCA2/genética , Análise Mutacional de DNA/métodos , Mutação , Neoplasias Ovarianas/patologia , Adulto , Idoso , Técnicas Citológicas/métodos , Feminino , Mutação em Linhagem Germinativa , Humanos , Pessoa de Meia-Idade , Neoplasias Ovarianas/diagnóstico , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo
9.
Appl Microbiol Biotechnol ; 103(11): 4443-4453, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30989251

RESUMO

The availability of preimmune libraries of antibody fragments allows for the fast generation of binders which can be expressed in both eukaryotic and prokaryotic systems. We exploited the recombinant nature of antibody fragments to demonstrate the possibility of expressing them as functional proteins displayed on the surface of Escherichia coli and by such a way to generate living reagents ready-to-use for diagnostics. Such immunoreagents were effectively exploited without the necessity of any purification step to prepare immunocapture surfaces suitable for the diagnostic of both cancer cells and toxic microalgae. The same nanobody-displaying bacteria were also engineered to coexpress GFP in their cytoplasm. Suspensions of such living fluorescent immunoreagents effectively bound to eukaryotic cells making them visible and quantifiable by flow cytometry analysis and using 96-well plate readers. The collected data showed the suitability of such living immunoreagents for reproducible and inexpensive diagnostic applications.


Assuntos
Técnicas de Visualização da Superfície Celular/métodos , Técnicas Citológicas/métodos , Escherichia coli/metabolismo , Proteínas Imobilizadas/metabolismo , Fatores Imunológicos/metabolismo , Proteínas Recombinantes/metabolismo , Anticorpos de Domínio Único/metabolismo , Aderência Bacteriana , Escherichia coli/genética , Proteínas Imobilizadas/genética , Imunoensaio/métodos , Fatores Imunológicos/genética , Proteínas Recombinantes/genética , Anticorpos de Domínio Único/genética , Coloração e Rotulagem/métodos
10.
Methods Cell Biol ; 151: 159-176, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30948006

RESUMO

Recent progress in multiplex cis-regulatory analysis has increased the speed of identifying enhancers and promoters, and enabled efficient incorporation of cis-regulatory information into gene regulatory network models. Three types of barcode reporters have been developed for multiplex reporter assays in sea urchin embryos: 13-tags and 129-tags for QPCR, 130 Nanotags for NanoString, and 100 million N25-tags for next-generation sequencing. In this chapter, to facilitate adoption of high-throughput cis-regulatory analysis in sea urchin embryos, I provide practical guidelines to best utilize barcode reporters that are compatible with either QPCR or next-generation sequencing. I expect that the guidelines are also applicable to other invertebrate embryos.


Assuntos
Técnicas Citológicas/métodos , Redes Reguladoras de Genes/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Análise de Sequência de DNA/métodos , Animais , Embrião não Mamífero , Regulação da Expressão Gênica no Desenvolvimento/genética , Regiões Promotoras Genéticas , Ouriços-do-Mar/genética
11.
Methods Cell Biol ; 151: 283-304, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30948012

RESUMO

The exquisite synchronicity of sea urchin development provides a reliable model for studying maternal proteins in the haploid egg as well as those involved in egg activation, fertilization and early development. Sea urchin eggs are released by the millions, enabling the quantitative evaluation of maternally stored and newly synthesized proteins over a range of time (seconds to hours post fertilization). During this window of development exist many hallmark and unique biochemical interactions that can be investigated for the purpose of characterizing profiles of kinases and other signaling proteins, manipulated using pharmacology to test sufficiency and necessity, for identification of post translational modifications, and for capturing protein-protein interactions. Coupled with the fact that sea urchin eggs and embryos are transparent, this synchronicity also results in large populations of cells that can be evaluated for newly synthesized protein localization and identification through use of the Click-iT technology. We provide basic protocols for these approaches and direct readers to the appropriate literature for variations and examples.


Assuntos
Biologia Celular/tendências , Técnicas Citológicas/métodos , Regulação da Expressão Gênica no Desenvolvimento/genética , Proteínas/efeitos adversos , Animais , Embrião não Mamífero , Desenvolvimento Embrionário/genética , Óvulo/crescimento & desenvolvimento , Óvulo/metabolismo , Biossíntese de Proteínas/genética , Proteínas/genética , Ouriços-do-Mar/genética , Ouriços-do-Mar/crescimento & desenvolvimento
13.
Genes Immun ; 20(5): 426-435, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31019256

RESUMO

Only a profound understanding of the structure and function of cells-either as single units or in the context of tissues and whole organisms-will allow a comprehension of what happens in pathological conditions and provides the means to fight disease. The Cell Biology and Infection (BCI for Biologie Cellulaire et Infection) department was created in 2002 at the Institut Pasteur in Paris to develop a research program under the umbrella of cell biology, infection biology, and microbiology. Its visionary ambition was to shape a common framework for cellular microbiology, and to interface the latter with hard sciences like physics and mathematics and cutting-edge technology. This concept, ahead of time, has given high visibility to the field of cellular microbiology and quantitative cell biology, and it has allowed the successful execution of highly interdisciplinary research programs linking a molecular understanding of cellular events with disease. Now, the BCI department embraces additional pathologies, namely cancer and neurodegenerative diseases. Here, we will portray how the integrative research approach of BCI has led to major scientific breakthroughs during the last 10 years, and where we see scientific opportunities for the near future.


Assuntos
Técnicas Citológicas/métodos , Infectologia/métodos , Pesquisa Interdisciplinar/métodos , França
14.
Soft Matter ; 15(21): 4266-4275, 2019 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-30968924

RESUMO

Functional cilia and flagella are crucial to the propulsion of physiological fluids, motile cells, and microorganisms. Motility assessment of individual cells allows discrimination of normal from dysfunctional behavior, but cell-scale analysis of individual trajectories to represent a population is laborious and impractical for clinical, industrial, and even research applications. We introduce an assay that quantifies swimming capability as a function of the variation in polar moment of inertia of cells released from an acoustic trap. Acoustic confinement eliminates the need to trace discrete trajectories and enables automated analysis of hundreds of cells in minutes. The approach closely approximates the average speed estimated from the mean squared displacement of individual cells for wild-type Chlamydomonas reinhardtii and two mutants (ida3 and oda5) that display aberrant swimming behaviors. Large-population acoustic trap-and-release rapidly differentiates these cell types based on intrinsic motility, which provides a highly sensitive and efficient alternative to conventional particle tracing.


Assuntos
Acústica , Chlamydomonas reinhardtii/citologia , Técnicas Citológicas/métodos , Chlamydomonas reinhardtii/genética , Cílios/metabolismo , Análise de Elementos Finitos , Flagelos/metabolismo , Mutação , Fatores de Tempo
15.
APMIS ; 127(4): 196-201, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30815926

RESUMO

The aim of this study was to assess the role of cytology, human papilloma virus (HPV) DNA and human papilloma virus messenger RNA (HPV mRNA) assays in detecting cervical intraepithelial neoplasia grade 2+ (CNi 2+) (recurrences/persistence) during the follow-up of women after treatment of cervical intraepithelial lesion. This cross-sectional study was performed among 43 women treated for cervical intraepithelial neoplasia (CIN) between January 2014 and January 2017 at the Department of Obstetrics and Gynecology of Spedali Civili's Hospital, Brescia, Italy. Pap smear and cervical samples for HPV tests were collected during the follow-up visit. Furthermore, colposcopy was always performed in order to find out the persistence/recurrence of the disease. A cervical biopsy was collected when necessary. Cervical samples obtained were tested for HPV DNA using the INNO-LiPa HPV assay and for HPV mRNA using the APTIMA assay. The mean age of enrolled women was 42.5 years. Among the treated patients, more than 50% of women revealed the absence of high risk HPV DNA and HPV mRNA. We found the persistence of the disease cervical intraepithelial neoplasia grade 2 (CIN 2) only in one woman. The sensitivity of cytology, HPV DNA and HPV mRNA in detecting disease was satisfactory (100%), while the specificity was quite different for the three tests: 64.2, 52.4 and 78.9%, respectively. The HPV mRNA test has higher specificity with respect to cytology and HPV DNA, avoiding the referral to unnecessary colposcopy with an improvement of costs/benefits for healthcare system. However, given the small size sample, this study should be considered as a pilot for future larger studies.


Assuntos
Neoplasia Intraepitelial Cervical/diagnóstico , Neoplasia Intraepitelial Cervical/patologia , Técnicas Citológicas/métodos , Testes Diagnósticos de Rotina/métodos , Técnicas de Diagnóstico Molecular/métodos , Infecções por Papillomavirus/complicações , Adulto , Idoso , DNA Viral/análise , Feminino , Seguimentos , Humanos , Itália , Pessoa de Meia-Idade , RNA Mensageiro/análise , RNA Viral/análise , Sensibilidade e Especificidade , Adulto Jovem
16.
Acta Cytol ; 63(3): 240-246, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30897562

RESUMO

OBJECTIVES: The diagnostic performance of cytology in esophageal squamous cell carcinoma (ESCC) is meticulously described. METHODS: Cytological and biopsy specimens were prospectively taken during esophagogastroduodenoscopy of 123 individuals in 2013 and 2014. Cytology samples were maintained in preservative fluid until processing and biopsies were formalin-fixed and paraffin-embedded. RESULTS: Based on endoscopic biopsy results, 70 cases were positive for ESCC whilst 53 were negative for cancer. In addition, brush cytology showed high sensitivity and specificity (98.57 and 96.23%, respectively) in detecting the disease, and high accuracy (97.5%) comparable to that provided by histopathology which is the accepted gold standard. CONCLUSION: Brush cytology specimens preserved in liquid medium may be a good alternative for ESCC diagnosis.


Assuntos
Biópsia/métodos , Citodiagnóstico/métodos , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/patologia , Esôfago/patologia , Estudos Transversais , Técnicas Citológicas/métodos , Neoplasias Esofágicas/diagnóstico , Carcinoma de Células Escamosas do Esôfago/diagnóstico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Conservantes Farmacêuticos , Estudos Prospectivos , Sensibilidade e Especificidade , Preservação de Tecido/métodos
17.
Spectrochim Acta A Mol Biomol Spectrosc ; 215: 297-302, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30844677

RESUMO

The design and synthesis of efficient probes for the detection of hydrogen sulfide is essential to explore the physiological role of such signaling molecules. A new probe,CDP containing 2-[(2,4-dinitrophenyl)thio]benzoic acid as responsive moiety was developed for H2S. After cleavage of responsive moiety occurs through a H2S-triggered nucleophilic reaction, the fluorescence of CDP was switched on. Up to 20-fold fluorescence enhancement toward H2S was observed and the detection limit was calculated to be as low as 307 nM. Moreover, this probe CDP was simple with good selectivity, high sensitivity and low cytotoxicity, which enabled it to detect H2S in solutions and exogenous/endogenous H2S in HepG-2 cells respectively.


Assuntos
Técnicas Citológicas/métodos , Corantes Fluorescentes/química , Sulfeto de Hidrogênio/química , Espectrometria de Fluorescência/métodos , Espectrofotometria Ultravioleta/métodos , Corantes Fluorescentes/metabolismo , Células Hep G2 , Humanos , Sulfeto de Hidrogênio/metabolismo , Modelos Lineares
18.
Vet Clin Pathol ; 48(1): 61-66, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30861155

RESUMO

BACKGROUND: The cytobrush technique is commonly used to sample the equine ocular surface. Impression cytology (IC) is an innovative noninvasive method, which allows for the collection of superficial layers of ocular epithelium. OBJECTIVES: The aims of this study were to compare the cytobrush and IC techniques on healthy equine ocular surfaces, to assess the agreement between observers with different levels of expertise, and to test the preservability of filters over time. METHODS: Twenty-four horses were sampled within 10 minutes of slaughter using IC on the left eye and the cytobrush technique on the right eye. May-Grünwald-Giemsa stained specimens were evaluated by two observers with different levels of expertise. Morphologic features were evaluated using a 4-grade system. The IC samples were re-evaluated after 6 months to examine filter preservation. RESULTS: In IC samples, corneal and conjunctival cells were clearly separated. Goblet cells were found in five and 17 filters by observer 1 and 2, respectively. Using the cytobrush technique, corneal and conjunctival cells were present but mixed. Goblet cell cellularity, preservation, and enumeration were higher with the IC technique compared with the cytobrush technique (P = 0.013; P = 0.004; P = 0.031, respectively). The inter-observer agreement for the IC technique was moderate to fair. In 7/24 IC samples re-evaluated after 6 months, cellular morphology was impaired, and the overall score was significantly lower. CONCLUSIONS: IC is an innovative noninvasive method, which allows for sample collection with higher cellularity and preservation. Moreover, the identification of goblet cells is easier. For these reasons, IC could be interesting and useful as a complementary diagnostic cytologic method in clinical practice.


Assuntos
Córnea/citologia , Técnicas Citológicas/veterinária , Cavalos/anatomia & histologia , Animais , Córnea/anatomia & histologia , Técnicas Citológicas/instrumentação , Técnicas Citológicas/métodos , Variações Dependentes do Observador , Manejo de Espécimes/veterinária
19.
Vet Clin Pathol ; 48(1): 143-147, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30861158

RESUMO

BACKGROUND: Sporotrichosis is an emerging zoonotic mycosis that presents as a cutaneous lymphatic or disseminated disease, caused by fungi from the Sporothrix schenkii (S schenkii) clinical clade. Its importance is growing, primarily due to an outbreak that occurred in Brazil, affecting mainly cats and people. OBJECTIVES: In Brazil, an S schenkii diagnosis is often made using cultures, which allows genus identification and sufficient growth to perform molecular biology testing. Despite its advantages, fungal cultures are slow to develop and can delay public health measures, highlighting the importance of developing additional diagnostics techniques. METHODS: Cell block cytology (CBLC) is an older method that regained importance after liquid-based cytology (LBC) was introduced, and it has been previously and successfully applied to veterinary diagnostics. We aimed to standardize and compare CBLC from cervical brush exfoliation of open wounds and fine-needle aspirates with culture and immunohistochemistry of skin biopsies for sporotrichosis in cats, as a novel method. RESULTS: For this purpose, we selected 40 cats with skin lesions suspected of having sporotrichosis in Guarulhos city, São Paulo state, Brazil. We achieved 97.5% and 95% positivity using CBLC and culture, respectively, and 100% of feline skin biopsies were positive for Sporothrix spp on histopathology/immunohistochemistry. CONCLUSIONS: Cell block cytology is an efficient and rapid tool to diagnose sporotrichosis in cats, particularly during epidemics.


Assuntos
Doenças do Gato/microbiologia , Dermatomicoses/veterinária , Técnicas de Preparação Histocitológica/veterinária , Sporothrix , Esporotricose/veterinária , Animais , Biópsia por Agulha Fina/veterinária , Doenças do Gato/diagnóstico , Doenças do Gato/patologia , Gatos , Técnicas Citológicas/instrumentação , Técnicas Citológicas/métodos , Técnicas Citológicas/veterinária , Dermatomicoses/diagnóstico , Dermatomicoses/microbiologia , Dermatomicoses/patologia , Feminino , Técnicas de Preparação Histocitológica/instrumentação , Técnicas de Preparação Histocitológica/métodos , Masculino , Técnicas Microbiológicas/métodos , Técnicas Microbiológicas/veterinária , Pele/citologia , Pele/microbiologia , Pele/patologia , Esporotricose/diagnóstico , Esporotricose/microbiologia , Esporotricose/patologia
20.
Methods Cell Biol ; 150: 269-292, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30777180

RESUMO

Small micromeres of the sea urchin are believed to be primordial germ cells (PGCs), fated to give rise to sperm or eggs in the adult. Sea urchin PGCs are formed at the fifth cleavage, undergo one additional division during blastulation, and migrate to the coelomic pouches of the pluteus larva. The goal of this chapter is to detail classical and modern techniques used to analyze primordial germ cell specification, gene expression programs, and cell behaviors in fixed and live embryos. The transparency of the sea urchin embryo enables both live imaging techniques and in situ RNA hybridization and immunolabeling for a detailed molecular characterization of these cells. Four approaches are presented to highlight small micromeres with fluorescent molecules for analysis by live and fixed cell microscopy: (1) small molecule dye accumulation during cleavage and blastula stages, (2) primordial germ cell targeted RNA expression using the Nanos untranslated regions, (3) fusing genes of interest with a Nanos2 targeting peptide, and (4) EdU and BrdU labeling. Applications of the live labeling techniques are discussed, including sorting by fluorescence-activated cell sorting for transcriptomic analysis, and, methods to image small micromere behavior in whole and dissociated embryos by live confocal microscopy. Finally, summary table of antibody and RNA probes as well as small molecule dyes to label small micromeres at a variety of developmental stages is provided.


Assuntos
Técnicas Citológicas/métodos , Células Germinativas/citologia , Ouriços-do-Mar/citologia , Animais , Embrião não Mamífero/citologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Larva/citologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA