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1.
Zootaxa ; 4790(3): zootaxa.4790.3.2, 2020 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-33056694

RESUMO

Slide mounted entomological specimens often require the aid of contrast techniques to improve the clarity of morphological characteristics. Methods can involve the use of techniques such as Phase contrast, Dark field or differential interference contrast microscopy (DIC), however where an entomologist may only have access to simple brightfield microscopy chemical staining of the specimen may be used to improve contrast. For whole mounts of entomological specimens, a single stain, occasionally two, are often used, in comparison to histological sections that often employ multiple stains in complex protocols. A number of authors have proposed different stains and staining methods for a number of insect groups with few considering the long term qualities of the stain, it has previously been shown that aniline dyes are prone to fading in Canada Balsam mounts, and that some stains fade even when protected from sunlight. This paper aims to summarise the knowledge of stains used for entomological specimens and provide details on the archival qualities.


Assuntos
Corantes , Insetos , Microscopia , Animais , Coloração e Rotulagem
2.
Int J Esthet Dent ; 15(4): 490-502, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33089262

RESUMO

AIM: This study assessed the changes in color and translucency of dentin and enamel after staining and bleaching procedures as well as the contribution of the tissues to color change (CTCC). MATERIALS AND METHODS: Fifty enamel-dentin specimens were obtained from bovine incisors and grouped into 10 blocks based on the similarity of the baseline color. One specimen in each block was used as the control, and the others were intrinsically (blood) or extrinsically (coffee) stained. One stained specimen for each pigment was bleached with 35% hydrogen peroxide. The enamel was separated from the dentin, and the tooth color and translucency parameter (TP) were measured individually and after recombining these substrates. Changes in color (ΔE00), whitening index (WI), and TP caused by the staining and bleaching procedures were estimated through assessing the color differences compared with the controls. CTCC was also calculated by recombining the enamel and dentin tissues from different treatments. The data were analyzed using the paired t test or the two-way repeated measures ANOVA (α = 0.05). RESULTS: Extrinsic staining procedures produced higher color changes than intrinsic ones, except in dentin. The highest bleaching effect was observed in the dentin of specimens that were extrinsically stained. Similar CTCC values were observed for the dentin and enamel for both the staining and bleaching procedures. Except for the specimens that were treated with intrinsic staining and became more translucent after the bleaching procedure, the other treatments resulted in non-significant TP changes. CONCLUSIONS: Staining the specimens extrinsically resulted in higher color change than staining them intrinsically, and these specimens were more affected by the bleaching procedure. Enamel and dentin demonstrated similar CTCC values.


Assuntos
Clareamento Dental , Animais , Bovinos , Esmalte Dentário , Dentina , Peróxido de Hidrogênio , Coloração e Rotulagem , Clareamento Dental/efeitos adversos
3.
PLoS One ; 15(8): e0237748, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32866195

RESUMO

Soil microbiota are considered a source of undiscovered bioactive compounds, yet cultivation of most bacteria within a sample remains generally unsuccessful. Two main reasons behind the unculturability of bacteria are the presence of cells in a viable but not culturable state (such as dormant cells) and the failure to provide the necessary growth requirements in vitro (leading to the classification of some bacterial taxa as yet-to-be-cultured). The present work focuses on the development of a single procedure that helps distinguish between both phenomena of unculturability based on viability staining coupled with flow cytometry and fluorescence-activated cell sorting. In the selected soil sample, the success rate of cultured bacteria was doubled by selecting viable and metabolically active bacteria. It was determined that most of the uncultured fraction was not dormant or dead but likely required different growth conditions. It was also determined that the staining process introduced changes in the taxonomic composition of the outgrown bacterial biomass, which should be considered for further developments. This research shows the potential of flow cytometry and fluorescence-activated cell sorting applied to soil samples to improve the success rate of bacterial cultivation by estimating the proportion of dormant and yet-to-be-cultured bacteria and by directly excluding dormant cells from being inoculated into growth media.


Assuntos
Bactérias/crescimento & desenvolvimento , Técnicas Microbiológicas/métodos , Microbiota/fisiologia , Microbiologia do Solo , Bactérias/química , Bactérias/genética , Biomassa , Separação Celular/métodos , Meios de Cultura , DNA Bacteriano/isolamento & purificação , Estudos de Viabilidade , Citometria de Fluxo/métodos , Corantes Fluorescentes/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Coloração e Rotulagem/métodos
4.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 42(4): 566-569, 2020 Aug 30.
Artigo em Chinês | MEDLINE | ID: mdl-32895113

RESUMO

The ganglioneuroma is a benign tumor originating from sympathetic ganglion cells.It often locates in the posterior mediastinum,retroperitoneum,and adrenal medulla.The intraspinal ganglioneuromas is relatively rare in clinical practice,which mainly locates in the cervical and thoracolumbar segments.A patient with main symptom of cough was examined by magnetic resonance imaging before operation in our center.Intraspinal ganglioneuromas was confirmed in the left intervertebral cavity area.Total resection of the tumor via the posterior median approach was performed.HE staining showed the mature ganglion cells were scattered.The patient was followed up for three months and no tumor recurrence occured.


Assuntos
Tosse/etiologia , Ganglioneuroma , Ganglioneuroma/complicações , Humanos , Imagem por Ressonância Magnética , Neurônios , Coloração e Rotulagem
5.
Tokai J Exp Clin Med ; 45(3): 148-151, 2020 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-32901905

RESUMO

PURPOSE: Pheochromocytoma (PCC) and paraganglioma (PGL) associated with the succinate dehydrogenase (SDH) germline mutations are characterized by negative results of immunohistochemistry tests for SDH subunit B (SDHB). Genetic testing for the SDH complex (SDHA, SDHB, SDHC, SDHD, and SDHAF2) is indicated only in patients with those diseases in whom immunohistochemistry tests for SDHB as a surrogate marker to detect the SDH complex mutation yield negative results. Two novel SDHB germline mutations, L157X and P236S, in PGL were previously reported. We therefore examined immunohistochemistry testing for SDHB in the PGLs with the SDHB germline mutations of L157X and P236S. METHODS: Immunohistochemistry for SDHB was performed in PGLs with the SDHB germline mutations of L157X and P236S. Five cases of sporadic PCC were subject to immunohistochemistry testing for SDHB. Normal tissue from the adrenal cortex adjacent to the sporadic PCC was used as the external positive control. RESULTS: Immunohistochemistry results were positive for SDHB in PGLs with the SDHB germline mutation of L157X and P236S, all five cases of sporadic PCC, and the adrenal cortex as the external positive control. CONCLUSION: Immunohistochemistry tests for SDHB showed positivity in PGLs associated with the SDHB germline mutations of L157X and P236S. Thus, immunohistochemistry testing for SDHB might not always reveal a surrogate marker in formal genetic testing of the SDH complex.


Assuntos
Neoplasias das Glândulas Suprarrenais/genética , Biomarcadores Tumorais/metabolismo , Mutação em Linhagem Germinativa , Paraganglioma/genética , Coloração e Rotulagem/métodos , Succinato Desidrogenase/genética , Succinato Desidrogenase/metabolismo , Córtex Suprarrenal/metabolismo , Humanos , Imuno-Histoquímica , Resultados Negativos
6.
Croat Med J ; 61(4): 354-365, 2020 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-32881434

RESUMO

AIM: To analyze axon morphology on rapid Golgi impregnated pyramidal neurons in the dorsolateral prefrontal cortex in schizophrenia. METHODS: Postmortem brain tissue from five subjects diagnosed with schizophrenia and five control subjects without neuropathological findings was processed with the rapid Golgi method. Layer III and layer V pyramidal neurons from Brodmann area 9 were chosen in each brain for reconstruction with Neurolucida software. The axons and cell bodies of 136 neurons from subjects with schizophrenia and of 165 neurons from control subjects were traced. The data obtained by quantitative analysis were compared between the schizophrenia and control group with the t test. RESULTS: Axon impregnation length was consistently greater in the schizophrenia group. The axon main trunk length was significantly greater in the schizophrenia than in the control group (93.7 ± 36.6 µm vs 49.8 ± 9.9 µm, P = 0.032). Furthermore, in the schizophrenia group more axons had visibly stained collaterals (14.7% vs 5.5%). CONCLUSION: Axon rapid Golgi impregnation stops at the beginning of the myelin sheath. The increased axonal staining in the schizophrenia group could, therefore, be explained by reduced axon myelination. Such a decrease in axon myelination is in line with both the disconnection hypothesis and the two-hit model of schizophrenia as a neurodevelopmental disease. Our results support that the cortical circuitry disorganization in schizophrenia might be caused by functional alterations of two major classes of principal neurons due to altered oligodendrocyte development.


Assuntos
Axônios/patologia , Córtex Pré-Frontal/patologia , Células Piramidais/patologia , Esquizofrenia/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Complexo de Golgi/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Coloração e Rotulagem/métodos
7.
PLoS One ; 15(9): e0235370, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32915779

RESUMO

Controlled experiments are one approach to understanding the pathogenicity of etiologic agents to susceptible hosts. The recently discovered fungal pathogen, Batrachochytrium salamandrivorans (Bsal), has resulted in a surge of experimental investigations because of its potential to impact global salamander biodiversity. However, variation in experimental methodologies could thwart knowledge advancement by introducing confounding factors that make comparisons difficult among studies. Thus, our objective was to evaluate if variation in experimental methods changed inferences made on the pathogenicity of Bsal. We tested whether passage duration of Bsal culture, exposure method of the host to Bsal (water bath vs. skin inoculation), Bsal culturing method (liquid vs. plated), host husbandry conditions (aquatic vs. terrestrial), and skin swabbing frequency influenced diseased-induced mortality in a susceptible host species, the eastern newt (Notophthalmus viridescens). We found that disease-induced mortality was faster for eastern newts when exposed to a low passage isolate, when newts were housed in terrestrial environments, and if exposure to zoospores occurred via water bath. We did not detect differences in disease-induced mortality between culturing methods or swabbing frequencies. Our results illustrate the need to standardize methods among Bsal experiments. We provide suggestions for future Bsal experiments in the context of hypothesis testing and discuss the ecological implications of our results.


Assuntos
Quitridiomicetos/patogenicidade , Micoses/veterinária , Urodelos/microbiologia , Animais , Técnicas de Cultura de Células , Quitridiomicetos/isolamento & purificação , Quitridiomicetos/fisiologia , Micoses/microbiologia , Micoses/patologia , Pele/microbiologia , Pele/patologia , Coloração e Rotulagem , Urodelos/fisiologia
9.
Wiad Lek ; 73(4): 789-791, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32731718

RESUMO

OBJECTIVE: The aim of the study to evaluate the peculiarities of the aortic wall structure at the place of coarctation. PATIENTS AND METHODS: Materials and methods: Studying of the aortic sections removed during operative correction at the place of constriction. 10 children at the age between 1 to 6 months were undergone the operation. Intraoperative aortic biopsy specimens were observed in 10% neutral formalin. Histologic sections were prepared in a conventional way followed by staining them with hematoxylin-eosin. RESULTS: Results: Histological examination in the areas of constriction revealed that the endothelium in all the preparations had poor expressiveness. The most significant changes were recorded in the middle layer of the aorta in the form of reduced development of elastic fibers, their fragmentation and chaotic arrangement. Angiomatosis with the formation of thin-layer small vessels by capillary type was found out. In all the preparations, areas of emptying of cells and fibers of the middle cover with the formation of cystic structures were revealed. CONCLUSION: Conclusions: The histological examination has revealed changes in the structure of the aorta wall, which may indicate the systemic nature of the lesion and make it possible to consider coarctation of the aorta to be a manifestation of systemic vasculopathy. The above-mentioned facts determine the need for a more detailed examination of children with the specified pathology at different stages of observation.


Assuntos
Coartação Aórtica , Aorta , Constrição Patológica , Técnicas Histológicas , Humanos , Lactente , Coloração e Rotulagem
10.
Artigo em Chinês | MEDLINE | ID: mdl-32791637

RESUMO

Objective:To evaluate the value of iodine staining in the margin determination in transoral laser microsurgery for early glottic carcinoma. Method:35 patients with early glottic laryngeal cancer(Tis-T2) were enrolled in this study from January 2016 to December 2017.All patients were applied with 1.5% Lugol iodine solution to vocal cords for 3 times after general anesthesia,and the coloration of the laryngeal cavity was observed under the operation microscope to determine the lesion margins.The lesions were excised completely with CO2 laser.The complications,complete resection rate and recurrence rate were evaluated. Result:In all patients,iodine staining was used to assist in determining the lesion range during transoral laser microsurgery.None of the patients had complications.Postoperative pathological results showed that the complete resection rate was 88.6%.There were no local recurrence cases in Tis and T1a,while local recurrence rate was 12.5% in T1b and 16.7% in T2,respectively.The overall local recurrence rate was 5.7%. Conclusion:The application of iodine staining in transoral laser microsurgery for early glottic carcinoma may help improve the complete resection rate and local control rate.It is a reliable,simple and low-cost method,worthy of further clinical application.Key words: laryngeal neoplasms;glottis; transoral laser microsurgery; iodine staining.


Assuntos
Carcinoma de Células Escamosas , Iodo , Neoplasias Laríngeas , Terapia a Laser , Glote , Humanos , Microcirurgia , Recidiva Local de Neoplasia , Estadiamento de Neoplasias , Estudos Retrospectivos , Coloração e Rotulagem , Resultado do Tratamento
11.
Zhonghua Yi Xue Za Zhi ; 100(30): 2372-2377, 2020 Aug 11.
Artigo em Chinês | MEDLINE | ID: mdl-32791814

RESUMO

Objective: To investigate the effects of serum immunoglobulin A/complement factor 3 (IgA/C3) ratio and glomerular C3 staining on clinical prognosis in patients with IgA nephropathy. Methods: From January 1st, 2007 to December 30th, 2016, a total of 519 patients with biopsy-proven IgA nephropathy (IgAN) in West China Hospital were retrospectively reviewed and divided into four groups based on serum IgA/C3 ratio and glomerular C3 staining: group A with IgA/C3 ratio ≥3.046 (median) and glomerular C3 staining ≥2 (n=151), group B with IgA/C3 ratio ≥3.046 and glomerular C3 staining<2 (n=109), group C with IgA/C3 ratio<3.046 and glomerular C3 staining ≥2 (n=119), and group D with IgA/C3 ratio<3.046 and glomerular C3 staining<2 (n=140). Clinical data, pathological characteristics and the primary endpoint [≥ 50% decline in estimated glomerular filtration rate (eGFR) and/or end-stage renal disease (ESRD)]were collected. Clinical prognosis and relevant risk factors were analyzed among the four groups. Results: Totally, 519 patients (298 males, 57.4%) with an average age of (33.6±10.9) years were recruited and followed up for (43.4±21.6) months. The rate of complete remission plus partial remission was 74.2% (112/151), 74.3% (81/109), 72.3% (86/119), 81.4% (114/140) in group A, B, C, D, respectively. Meanwhile, The rate of ESRD was highest in group A (14.6% vs 9.2%, 13.4%, 8.6%). Renal outcome (patients reached the endpoint) was worse in group A and C compared with group B and D (15.2%, 16.0 vs 8.3%, 7.9%). Moreover, 80-month renal survival rate was significantly worse in group A (84.8%) than that in group B and D (91.7% and 92.1%), but no statistical significant difference was found between group A and B (P(AB)=0.085; P(AD)=0.028). There was no significant difference of renal survival rate between group A and C (84.8% vs 84.0%, P=0.896). Multivariate Cox model showed that hypertension (HR=2.753, 95%CI: 1.452-5.217, P=0.002), serum creatinine (HR=1.011, 95%CI: 1.008-1.014, P<0.001), and tubular atrophy/interstitial fibrosis (T1/T2) (HR=6.595, 95%CI: 3.107-13.999, P<0.001) were independent predictors of poor renal survival. Conclusion: Serum IgA/C3 ratio and glomerular C3 staining are predictors of renal clinical prognosis in patients with IgA nephropathy.


Assuntos
Glomerulonefrite por IGA , Adulto , China , Complemento C3/análise , Progressão da Doença , Taxa de Filtração Glomerular , Humanos , Imunoglobulina A , Masculino , Prognóstico , Estudos Retrospectivos , Coloração e Rotulagem , Adulto Jovem
12.
Nat Protoc ; 15(9): 2813-2836, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32747820

RESUMO

Several noncanonical initial nucleotides (NCINs) have been found to cap RNAs and possibly regulate RNA stability, transcription and translation. NAD+ is one of the NCINs that has recently been discovered to cap RNAs in a wide range of species. Identification of the NAD+-capped RNAs (NAD-RNAs) could help to unveil the cap-mediated regulation mechanisms. We previously reported a method termed NAD tagSeq for genome-wide analysis of NAD-RNAs. NAD tagSeq is based on the previously published NAD captureSeq protocol, which applies an enzymatic reaction and a click chemistry reaction to label NAD-RNAs with biotin followed by enrichment with streptavidin resin and identification by RNA sequencing. In the current NAD tagSeq method, NAD-RNAs are labeled with a synthetic RNA tag and identified by direct RNA sequencing based on Oxford Nanopore technology. Compared to NAD captureSeq, NAD tagSeq provides a simpler procedure for direct sequencing of NAD-RNAs and noncapped RNAs and affords information on the whole sequence organization of NAD-RNAs and the ratio of NAD-RNAs to total transcripts. Furthermore, NAD-RNAs can be enriched by hybridizing a complementary DNA probe to the RNA tag, thus increasing the sequencing coverage of NAD-RNAs. The strategy of tagging RNAs with a synthetic RNA tag and identifying them by direct RNA sequencing might be employed in analyzing other NCIN-capped RNAs. The experimental procedure of NAD tagSeq, including RNA extraction, RNA tagging and direct RNA sequencing, takes ~5 d, and initial data analysis can be completed within 2 d.


Assuntos
Perfilação da Expressão Gênica , NAD/metabolismo , Capuzes de RNA/genética , Capuzes de RNA/metabolismo , Análise de Sequência de RNA/métodos , Coloração e Rotulagem
13.
Exp Parasitol ; 217: 107938, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32768560

RESUMO

PURPOSE: Praziquantel (PZQ) is the conventional antibilharzial agent. Nevertheless, no antibilharzial prophylactic agents or 100% curable therapy approved and no reported data about use of human CD34+ Umbilical Cord Blood Stem Cells (CD34+UCBSCs) or Wharton Jelly Mesenchymal Stem Cells (WJMSCs) in prevention and/or complete eradication of acute S.mansoni granulomas in liver. We aimed to study possible prophylactic vs therapeutic role of human CD34+UCBSCs and WJMSCs in acute hepatic bilharzial granulomas in pre vs post-infected mice. METHODS: Seventy mice were divided into 7 groups (10 mice each): Normal, S.mansoni-infected, post-infected PZQ-treated, CD34+UCBSCs pre and post-infected, WJMSCs pre and post-infected. Serological, parasitological, histopathological evaluation using OCT4 & TGFB immunohistochemistry and quantitative image analysis assessment of TGFB-stained fibrogenesis in liver granulomas performed. RESULTS: Histopathologically, surprisingly and significantly, the prophylactic pre-infection stem cells (CD34+UCBSCs and WJMSCs) & similarly the post-infection CD34+UCBSCs treatment revealed eradication/reversal of the entire granulomas and no fibrosis. Moreover, post-infection PZQ treatment showed fewer and significantly smaller granulomas than post-infection WJMSCs treatment. Nevertheless, post-infection WJMSCs exhibited non-significant less TGFB-stained fibrogenesis. CONCLUSION: CD34+UCBSCs exerted the best prophylactic and therapeutic roles in prevention and complete cure of acute hepatic S.mansoni granulomas over WJMSCs and PZQ. In contrast, only pre-infection WJMSCs exhibited similar preventive (prophylactic) effect. On the contrary, post-infection WJMSCs were the worst (incompletely reversed granulomas). Post-infection Praziquantel was overall better therapeutically than WJMSCs in this regard. Accordingly, when it comes to WJMSCs application, WJMSCs are better used as a pre-infection prophylactic and preventive tool rather than a post-infection therapy. Further studies are needed.


Assuntos
Antígenos CD34/sangue , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Esquistossomose mansoni/prevenção & controle , Esquistossomose mansoni/terapia , Animais , Anti-Helmínticos/administração & dosagem , Fezes/parasitologia , Sangue Fetal/citologia , Citometria de Fluxo , Granuloma/prevenção & controle , Granuloma/terapia , Humanos , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Fígado/patologia , Hepatopatias Parasitárias/prevenção & controle , Hepatopatias Parasitárias/terapia , Masculino , Células-Tronco Mesenquimais , Camundongos , Fator 3 de Transcrição de Octâmero , Contagem de Ovos de Parasitas , Praziquantel/administração & dosagem , Coloração e Rotulagem , Fator de Crescimento Transformador beta
14.
Zhonghua Yan Ke Za Zhi ; 56(6): 465-472, 2020 Jun 11.
Artigo em Chinês | MEDLINE | ID: mdl-32842329

RESUMO

Objective: To observe the clinical effect of corneal staining in patients with corneal leucoma. Methods: Restrospective case series study.Thirty eyes of 30 patients (17 males, 13 females) with corneal leucoma who underwent corneal interlamellar staining at Beijing Aier-Intech Eye Hospital from October 2014 to July 2018 were included. The mean age was 31.50±15.66 years. Postoperative follow-up was more than 1 year. All the patients underwent appearance examination, anterior segment examination with a slit lamp, B-scan examination and anterior segment OCT examination preoperatively and postoperatively. The effect of corneal staining, patient satisfaction and the incidence of complications were investigated. Results: The corneal epithelium of all the patients healed completely within 1-3 days after operation. The mean follow-up period was 30.68±18.02 months (range, 12.37-58.10 months). During the postoperative follow-up period, no staining permeation or inflammatory reaction in the anterior chamber occurred. The corneal color and appearance were well maintained in 18 patients (72.0%). Seven patients (28.0%) showed mild corneal color-fading. One patient was treated with enucleation and orbital hydroxyapatite implantation because of eye atrophy and corneal banding degeneration caused by the primary disease at 4 years after operation. The surgeon was satisfied with the improvement of the appearance of all cases; 22 patients (88.0%) were satisfied with the postoperative appearance. No significant complications were observed in all the cases. Conclusions: Corneal interlamellar staining is one of best choices for the treatment of corneal leucoma. It has advantages of quick postoperative recovery, long-standing color staining and good cosmetic effect. The operation is simple and easy to carry out and there is no obvious damage to eye tissues. (Chin J Ophthalmol, 2020, 56:465-472).


Assuntos
Opacidade da Córnea , Cosméticos , Adolescente , Adulto , Córnea , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Coloração e Rotulagem , Adulto Jovem
15.
J Environ Manage ; 275: 111249, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32836169

RESUMO

Lagooning sludge (LS), which is used as soil amendment in Morocco, may contain microplastics (MPs). The aim of this study was to examine the effect of dewatering and co-composting of LS with green waste (GW) on the MPs' evolution. In this context the present study proposes fast-preliminary steps to detect plastics in lagooning sewage sludge before the extraction and identification process. We used pyrolysis GC/MS spectrometry to investigate the presence of chemical compounds possibly derived from plastics, and fluorescence staining by Nile Red to detect fluorescent particles suspected as plastics. Thereafter, we quantified the MPs particles after density fractionation and investigated their nature by Raman spectroscopy. RESULTS: indicated the presence of an average of 40.5 ± 11.9 × 103 MPs particles/kg (dry matter) and 36 ± 9.7 × 103 MPs particles/kg (dry matter) in fresh sludge and dewatered sludge respectively. Sludge dewatering in drying beds resulted a loss of small MPs (<500 µm). In co-composts, the quantity of MPs varied with the proportion of sewage sludge. The distribution of MPs types differentiated by colour and types (polypropylene, polyethylene, polyamide and polyester) evolved differently. Conventional co-composting did not have any effect on MPs quantity, indicating that they are not biodegradable under these temperature conditions, but it influenced their particle size. The risks of these pollutants after repeated field application and the possibility of their reduction through others co-composting procedures and techniques would be further investigated.


Assuntos
Compostagem , Esgotos , Microplásticos , Marrocos , Plásticos , Pirólise , Análise Espectral Raman , Coloração e Rotulagem
16.
Nat Commun ; 11(1): 4339, 2020 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-32859909

RESUMO

DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) facilitates multiplexing in superresolution microscopy but is practically limited by slow imaging speed. To address this issue, we propose the additions of ethylene carbonate (EC) to the imaging buffer, sequence repeats to the docking strand, and a spacer between the docking strand and the affinity agent. Collectively termed DNA-PAINT-ERS (E = EC, R = Repeating sequence, and S = Spacer), these strategies can be easily integrated into current DNA-PAINT workflows for both accelerated imaging speed and improved image quality through optimized DNA hybridization kinetics and efficiency. We demonstrate the general applicability of DNA-PAINT-ERS for fast, multiplexed superresolution imaging using previously validated oligonucleotide constructs with slight modifications.


Assuntos
Técnicas Citológicas/métodos , DNA/química , Microscopia de Fluorescência/métodos , Simulação de Acoplamento Molecular/métodos , Linhagem Celular , Humanos , Processamento de Imagem Assistida por Computador/métodos , Oligonucleotídeos , Coloração e Rotulagem/métodos
17.
Nat Commun ; 11(1): 3850, 2020 07 31.
Artigo em Inglês | MEDLINE | ID: mdl-32737322

RESUMO

Resolving the distribution of specific proteins at the nanoscale in the ultrastructural context of the cell is a major challenge in fluorescence microscopy. We report the discovery of a new principle for an optical contrast equivalent to electron microscopy (EM) which reveals the ultrastructural context of the cells with a conventional confocal microscope. By decrowding the intracellular space through 13 to 21-fold physical expansion while simultaneously retaining the proteins, bulk (pan) labeling of the proteome resolves local protein densities and reveals the cellular nanoarchitecture by standard light microscopy.


Assuntos
Imageamento Tridimensional/métodos , Microscopia de Fluorescência/métodos , Proteoma/análise , Coloração e Rotulagem/métodos , Acrilamidas/química , Reagentes para Ligações Cruzadas/química , Corantes Fluorescentes/química , Células HeLa , Humanos , Hidrogéis/química , Espaço Intracelular/química , Succinimidas/química , Inclusão do Tecido/métodos
19.
Rinsho Shinkeigaku ; 60(8): 565-568, 2020 Aug 07.
Artigo em Japonês | MEDLINE | ID: mdl-32641630

RESUMO

A 49-year-old woman was admitted to our hospital with suspected hypertensive encephalopathy. On the basis of MRI showing leptomeningeal enhancement and Class V cytology of the CSF, she was diagnosed as having leptomeningeal carcinomatosis. Although no primary site was detected, a few melanin granules were observed at the third CSF examination. The atypical cells in the CSF demonstrated immunoreactivity for HMB-45 and S-100, which are specific markers of malignant melanoma. There have been few reports of meningeal melanomatosis in Japan. This case illustrates that immunostaining is diagnostically useful in patients with leptomeningeal carcinomatosis from neoplasms with unknown primary sites.


Assuntos
Biomarcadores Tumorais/líquido cefalorraquidiano , Líquido Cefalorraquidiano , Citodiagnóstico/métodos , Melanoma/líquido cefalorraquidiano , Melanoma/diagnóstico , Carcinomatose Meníngea/líquido cefalorraquidiano , Carcinomatose Meníngea/diagnóstico , Neoplasias Meníngeas/líquido cefalorraquidiano , Neoplasias Meníngeas/diagnóstico , Coloração e Rotulagem/métodos , Idoso , Feminino , Humanos , Antígenos Específicos de Melanoma/líquido cefalorraquidiano , Neoplasias Primárias Desconhecidas , Proteínas S100/líquido cefalorraquidiano
20.
Nat Commun ; 11(1): 3284, 2020 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-32601292

RESUMO

The inner nuclear membrane (INM) selectively accumulates proteins that are essential for nuclear functions; however, overaccumulation of INM proteins results in a range of rare genetic disorders. So far, little is known about how defective, mislocalized, or abnormally accumulated membrane proteins are actively removed from the INM, especially in plants and animals. Here, via analysis of a proximity-labeling proteomic profile of INM-associated proteins in Arabidopsis, we identify critical components for an INM protein degradation pathway. We show that this pathway relies on the CDC48 complex for INM protein extraction and 26S proteasome for subsequent protein degradation. Moreover, we show that CDC48 at the INM may be regulated by a subgroup of PUX proteins, which determine the substrate specificity or affect the ATPase activity of CDC48. These PUX proteins specifically associate with the nucleoskeleton underneath the INM and physically interact with CDC48 proteins to negatively regulate INM protein degradation in plants.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de Membrana/metabolismo , Membrana Nuclear/metabolismo , Proteoma/metabolismo , Proteômica/métodos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Proteínas de Membrana/genética , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Membrana Nuclear/genética , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Plantas Geneticamente Modificadas , Complexo de Endopeptidases do Proteassoma/metabolismo , Ligação Proteica , Proteólise , Proteoma/genética , Coloração e Rotulagem/métodos , Espectrometria de Massas em Tandem/métodos , Proteína com Valosina/genética , Proteína com Valosina/metabolismo
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