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1.
Adv Exp Med Biol ; 1131: 7-26, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31646505

RESUMO

Measuring free Ca2+ concentration ([Ca2+]) in the cytosol or organelles is routine in many fields of research. The availability of membrane permeant forms of indicators coupled with the relative ease of transfecting cell lines with biological Ca2+ sensors have led to the situation where cellular and subcellular [Ca2+] is examined by many non-specialists. In this chapter, we evaluate the most used Ca2+ indicators and highlight what their major advantages and disadvantages are. We stress the potential pitfalls of non-ratiometric techniques for measuring Ca2+ and the clear advantages of ratiometric methods. Likely improvements and new directions for Ca2+ measurement are discussed.


Assuntos
Cálcio , Citosol , Organelas , Animais , Cálcio/metabolismo , Técnicas Citológicas , Citosol/química , Citosol/metabolismo , Humanos , Organelas/química , Organelas/metabolismo
2.
Vet Dermatol ; 30(4): 337-e94, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31038257

RESUMO

BACKGROUND: Otitis is common in alpacas. Suppurative otitis media/interna can be an extension from the external ear canal or from a respiratory infection. Cytological evaluation provides rapid and inexpensive information to assist in therapeutic decision; to date, there is no published information regarding the normal cytological results and flora of the alpaca external ear canal. HYPOTHESIS/OBJECTIVES: To describe normal resident cytological findings and flora and possible variation over time, we sampled clinically normal alpaca external ear canals during two different seasons. ANIMALS: Fifty privately owned, healthy alpacas of different ages and sexes in two northeastern United States flocks. METHODS AND MATERIALS: One ear per alpaca had both cytological swabs (ectoparasites, inflammatory and epithelial cells, bacteria and yeast) and sterile swabs (bacterial and fungal cultures) taken. This was done in August 2017 and repeated in January 2018. RESULTS: Yeast organisms were noted cytologically in 2-4% of the samples. Prevalence of total yeast genera was 6% in August and 30% in January. Cytologically, rod-shaped bacteria [maximum 4-10/high power field (HPF); median 0-0.5/HPF] were seen in 50% of alpacas in August and 26% in January. Coccal bacteria (maximum 6-10/HPF; median 0/HPF) were seen in 32% of alpacas in August and 16% in January. No statistically significant findings were noted between sampling months. Common bacterial genera isolated in August were Bacillus (44%), Arthrobacter (40%) and nonhaemolytic Staphylococcus (26%), and in January were Bacillus (42%) and Pantoea (38%). CONCLUSIONS AND CLINICAL IMPORTANCE: This information may be useful when evaluating alpaca external ear canal samples, which subsequently may help dictate empirical therapy.


Assuntos
Bactérias/isolamento & purificação , Técnicas Citológicas/veterinária , Meato Acústico Externo/microbiologia , Animais , Bactérias/classificação , Camelídeos Americanos , Feminino , Masculino , Estações do Ano , Manejo de Espécimes
3.
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
4.
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
5.
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
6.
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
7.
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
11.
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
12.
BMC Womens Health ; 19(1): 47, 2019 03 25.
Artigo em Inglês | MEDLINE | ID: mdl-30909894

RESUMO

BACKGROUND: European guidelines for cervical cancer screening now recommend the use of clinically validated assays for high-risk HPV-DNA sequences as primary test in women older than 30 years, performed in centralized laboratories, and run on systems providing automated solutions for all steps. METHODS: We conducted a comparison study, according to the international guidelines, nested within the organized population-based cervical screening program, between the cobas 4800 and 6800 systems (Roche Diagnostics), to evaluate accuracy and reproducibility of HPV test results and laboratory workflow. In Italy implementation of HPV cervical screening is under way on a regional basis; in Veneto it started in June 2015, following a piloting phase; the assay in use in the three centralized laboratories is the cobas 4800 HPV test, run on the cobas 4800 system. Comparison of HPV results with a new version of the assay (cobas 6800/8800 HPV) run on the cobas 6800 system, and intra- and inter-reproducibility analyses have been conducted in samples collected in PreservCyt medium (Hologic) from women without and with a subsequent diagnosis of high-grade lesion. RESULTS: Samples from women older than 30 years attending organized cervical cancer screening were used. Clinical sensitivity and specificity were evaluated on 60 cases and 925 controls, respectively; intra-laboratory reproducibility and inter-laboratory agreement by the 6800 system were evaluated on 593 and 460 specimens, respectively. Our results showed a very high agreement (> 98%) for overall qualitative results between the two systems; clinical sensitivity and specificity of the HPV assay run on 6800 were non-inferior to those of the HPV assay run on 4800 (p = 0,0157 and p = 0,0056, respectively, at the recommended thresholds of 90 and 98%); kappa values of 0.967 and 0.969 were obtained for intra- and inter-laboratory reproducibility analyses in the 6800 system. The 6800 platform displayed several technological improvements over the 4800 system, with higher throughput and laboratory productivity, and lower operator's hands-on time. CONCLUSIONS: The new cobas 6800/8800 HPV assay run on the 6800 instrument is suitable for use in large centralized laboratories included within population-based cervical cancer screening programs.


Assuntos
Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/normas , Neoplasias do Colo do Útero/diagnóstico , Carga Viral/métodos , Adulto , Técnicas Citológicas/normas , Detecção Precoce de Câncer/métodos , Feminino , Humanos , Itália , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular/instrumentação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
13.
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
14.
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
15.
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
16.
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
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.
Turk Patoloji Derg ; 35(2): 119-127, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30715725

RESUMO

OBJECTIVE: This study was designed to determine whether obesity causes the development of metaplasia in conjunctival epithelial cells. MATERIAL AND METHOD: A total of 61 volunteer participants who had no previous history of illness or drug use were involved in this study. Of those, 20 were obese, and 41 were of normal weight. We measured the glucose and insulin values of all volunteers. We also measured the Body Mass Index (BMI) and Homeostasis Model Assessment for Insulin Resistance (HOMA IR). The impression cytology method was used to analyze the conjunctival epithelium cells, and to classify them between Grades 0 to 3 according to the Nelson criteria. RESULTS: There was a certain level of loss of goblet cells on the 90% level as well as squamous metaplasia (Grade 2-3) in 80% of the obese participants and impression cytology was found to be normal in only two patients. The expected results were observed in 56.1% of the control group where the squamous metaplasia rate was nearly 17% (p < 0.001). 90.9% of the grade 3 patients were obese. The variables as independent predictors were found to indicate the existence of abnormal cytology in the conjunctiva at various levels; BMI (OR: 1.24; p=0.002) and HOMA IR (OR= 28.6; p= 0.001) in a Model I multivariable regression model, and the existence of obesity (OR: 11.91; p=0.002) and HOMA IR (OR= 15.08; p < 0.001) in a Model II multivariable regression model. CONCLUSION: Obesity was found to be a disorder that causes metaplasia in the conjunctival epithelium cells for the first time.


Assuntos
Túnica Conjuntiva/patologia , Resistência à Insulina , Obesidade/complicações , Adulto , Glicemia/análise , Índice de Massa Corporal , Túnica Conjuntiva/citologia , Técnicas Citológicas , Células Epiteliais/citologia , Células Epiteliais/patologia , Feminino , Células Caliciformes/citologia , Homeostase , Humanos , Insulina/sangue , Masculino , Glândulas Tarsais/fisiopatologia , Metaplasia/etiologia , Pessoa de Meia-Idade , Transtornos da Visão/etiologia
19.
Lab Chip ; 19(7): 1141-1152, 2019 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-30778467

RESUMO

Developing embryos create complexity by expressing genes to coordinate movement which generates mechanical force. An emerging theory is that mechanical force can also serve as an input signal to regulate developmental gene expression. Experimental methods to apply mechanical stimulation to whole embryos have been limited, mainly to aspiration, indentation, or moving a coverslip; these approaches stimulate only a few embryos at a time and require manual alignment. A powerful approach for automation is microfluidic devices, which can precisely manipulate hundreds of samples. However, using microfluidics to apply mechanical stimulation has been limited to small cellular systems, with fewer applications for larger scale whole embryos. We developed a mesofluidic device that applies the precision and automation of microfluidics to the Drosophila embryo: high-throughput automatic alignment, immobilization, compression, real-time imaging, and recovery of hundreds of live embryos. We then use twist:eGFP embryos to show that the mechanical induction of twist depends on the dose and duration of compression. This device allows us to quantify responses to compression, map the distribution of ectopic twist, and measure embryo stiffness. For building mesofluidic devices, we describe modifications on ultra-thick photolithography, derive an analytical model that predicts the deflection of sidewalls, and discuss parametric calibration. This "mesomechanics" approach combines the high-throughput automation and precision of microfluidics with the biological relevance of live embryos to examine mechanotransduction. These analytical models facilitate the design of future devices to process multicellular organisms such as larvae, organoids, and mesoscale tissue samples.


Assuntos
Técnicas Citológicas/instrumentação , Drosophila/embriologia , Embrião não Mamífero/citologia , Dispositivos Lab-On-A-Chip , Mecanotransdução Celular , Animais , Calibragem , Módulo de Elasticidade , Desenho de Equipamento
20.
Bull Tokyo Dent Coll ; 60(1): 29-37, 2019 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-30700642

RESUMO

Oral exfoliative cytology is now used by general practitioners in Japan to screen for oral cancer. With conventional cytology, however, the number of cells that can be sampled is small. Moreover, cell deformation and piling of cells when preparing specimens has been reported. The purpose of this study was to compare conventional and liquid based cytology (LBC), which has been employed with increasing frequency in recent years. We believe that identifying potential pitfalls in oral exfoliative cytology will help improve diagnostic accuracy. A total of 153 patients with tongue squamous cell carcinoma who were diagnosed and treated initially at our hospital between January 2000 and December 2010 were included. Of these, 124 underwent conventional cytology, while the remaining 29 underwent LBC. Histopathological and clinical findings were used as criteria. Conventional cytology yielded a positive rate of 54.8% and LBC 79.3%, while values of 28.2% and 13.8% were obtained for a suspected positive rate, respectively. Liquid based cytology yielded a significantly higher percentage of accurate diagnoses and fewer suspected positives (p<0.05) in cases clinically classified as endophytic and those classified as ulcerative in terms of clinical growth pattern. No significant difference was observed between conventional cytology and LBC in cases of an infiltrative growth pattern, however.The present results suggest that LBC is superior to conventional cytology in achieving an accurate diagnosis based on oral exfoliative cytology. The present findings also suggest that exophytic type, and especially leukoderma type clinical growth patterns constitute pitfall cases in oral exfoliative cytology.


Assuntos
Carcinoma de Células Escamosas/diagnóstico , Técnicas Citológicas/métodos , Detecção Precoce de Câncer/métodos , Neoplasias da Língua/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Adulto Jovem
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