scholarly journals The Moss Leptodictyum riparium Counteracts Severe Cadmium Stress by Activation of Glutathione Transferase and Phytochelatin Synthase, but Slightly by Phytochelatins

2020 ◽  
Vol 21 (5) ◽  
pp. 1583 ◽  
Author(s):  
Erika Bellini ◽  
Viviana Maresca ◽  
Camilla Betti ◽  
Monica Ruffini Castiglione ◽  
Debora Fontanini ◽  
...  
Keyword(s):  
High Performance ◽  
Laser Scanning ◽  
Glutathione Transferase ◽  
Cadmium Stress ◽  
Laser Scanning Microscopy ◽  
Cellular Damage ◽  
Confocal Laser ◽  
Peptidase Activity ◽  
Phytochelatin Synthase ◽  
Glutathione S Transferase

In the present work, we investigated the response to Cd in Leptodictyum riparium, a cosmopolitan moss (Bryophyta) that can accumulate higher amounts of metals than other plants, even angiosperms, with absence or slight apparent damage. High-performance liquid chromatography followed by electrospray ionization tandem mass spectrometry of extracts from L. riparium gametophytes, exposed to 0, 36 and 360 µM Cd for 7 days, revealed the presence of γ-glutamylcysteine (γ-EC), reduced glutathione (GSH), and traces of phytochelatins. The increase in Cd concentrations progressively augmented reactive oxygen species levels, with activation of both antioxidant (catalase and superoxide dismutase) and detoxifying (glutathione-S-transferase) enzymes. After Cd treatment, cytosolic and vacuolar localization of thiol peptides was performed by means of the fluorescent dye monochlorobimane and subsequent observation with confocal laser scanning microscopy. The cytosolic fluorescence observed with the highest Cd concentrations was also consistent with the formation of γ-EC-bimane in the cytosol, possibly catalyzed by the peptidase activity of the L. riparium phytochelatin synthase. On the whole, activation of phytochelatin synthase and glutathione-S-transferase, but minimally phytochelatin synthesis, play a role to counteract Cd toxicity in L. riparium, in this manner minimizing the cellular damage caused by the metal. This study strengthens previous investigations on the L. riparium ability to efficiently hinder metal pollution, hinting at a potential use for biomonitoring and phytoremediation purposes.

scholarly journals Polyamines Are Essential for the Formation of Plague Biofilm

2006 ◽  
Vol 188 (7) ◽  
pp. 2355-2363 ◽  
Author(s):  
Chandra N. Patel ◽  
Brian W. Wortham ◽  
J. Louise Lines ◽  
Jacqueline D. Fetherston ◽  
Robert D. Perry ◽  
...  
Keyword(s):  
High Performance ◽  
Laser Scanning ◽  
Double Mutant ◽  
Arginine Decarboxylase ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Crystal Violet Staining ◽  
Liquid Chromatography Mass Spectrometry ◽  
Double Deletion ◽  
Single Deletion

ABSTRACT We provide the first evidence for a link between polyamines and biofilm levels in Yersinia pestis, the causative agent of plague. Polyamine-deficient mutants of Y. pestis were generated with a single deletion in speA or speC and a double deletion mutant. The genes speA and speC code for the biosynthetic enzymes arginine decarboxylase and ornithine decarboxylase, respectively. The level of the polyamine putrescine compared to the parental speA + speC + strain (KIM6+) was depleted progressively, with the highest levels found in the Y. pestis ΔspeC mutant (55% reduction), followed by the ΔspeA mutant (95% reduction) and the ΔspeA ΔspeC mutant (>99% reduction). Spermidine, on the other hand, remained constant in the single mutants but was undetected in the double mutant. The growth rates of mutants with single deletions were not altered, while the ΔspeA ΔspeC mutant grew at 65% of the exponential growth rate of the speA + speC + strain. Biofilm levels were assayed by three independent measures: Congo red binding, crystal violet staining, and confocal laser scanning microscopy. The level of biofilm correlated to the level of putrescine as measured by high-performance liquid chromatography-mass spectrometry and as observed in a chemical complementation curve. Complementation of the ΔspeA ΔspeC mutant with speA showed nearly full recovery of biofilm to levels observed in the speA + speC + strain. Chemical complementation of the double mutant and recovery of the biofilm defect were only observed with the polyamine putrescine.

Research on Printed Conductive Silver Layer based on Inkjet Photo Paper

2021 ◽  
Author(s):  
Yi Fang ◽  
Lixin Mo ◽  
Zhiqing Xin ◽  
Yinjie Chen ◽  
Xiu Li ◽  
...  
Keyword(s):  
High Performance ◽  
Laser Scanning ◽  
Low Cost ◽  
Flexible Substrate ◽  
Electronic Devices ◽  
Laser Scanning Microscopy ◽  
Silver Layer ◽  
Confocal Laser ◽  
Nano Silver ◽  
Silver Ink

Printed electronics is an emerging technology that applies traditional printing or coating processes to the manufacture of electronic devices and products. In order to find a low-cost, high-performance, environmentally-friendly flexible substrate suitable for electronic devices, the printability between four kinds of inkjet photo papers and nano-silver ink was investigated. First, different surface morphologies of the inkjet photo papers were measured by a confocal laser scanning microscopy. Then, a pen and a gravure printer were used to test the printability between photo papers and nano-silver ink. It was found that the conductive track and pattern was influenced by the surface morphology of the photo papers. Furthermore, a four-probe test showed that the conductivity of the ink layers on the four photo papers was almost at the same level. Furthermore, a tearing test with 3 M tapes showed that the silk photo paper had the best tearing resistance. In general, silk photo paper has the best overall performance. This research could be beneficial for the development of flexible electronic devices which are low-cost, mass manufacture suitable and environment friendly.

Autophagy is required for sea urchin oogenesis and early development

Zygote ◽  
2016 ◽  
Vol 24 (6) ◽  
pp. 918-926 ◽  
Author(s):  
Maria Agnello ◽  
Roberto Chiarelli ◽  
Chiara Martino ◽  
Liana Bosco ◽  
Maria Carmela Roccheri
Keyword(s):  
Sea Urchin ◽  
Laser Scanning ◽  
Marine Invertebrates ◽  
Paracentrotus Lividus ◽  
Cadmium Stress ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Specific Localization ◽  
Developmental Programme

SummaryAutophagy is a major intracellular pathway for the degradation and recycling of cytosolic components. Emerging evidence has demonstrated its crucial role during the embryo development of invertebrates and vertebrates. We recently demonstrated a massive activation of autophagy in Paracentrotus lividus embryos under cadmium stress conditions, and the existence of a temporal relationship between induced autophagy and apoptosis. Although there have been numerous studies on the role of autophagy in the development of different organisms, information on the autophagic process during oogenesis or at the start of development in marine invertebrates is very limited. Here we report our recent data on the occurrence of autophagy at these key phases of development. In order to investigate autophagy trends we performed in vivo assays to detect autophagolysomes, as well as in situ analysis with anti-LC3 antibody to detect autophagosomes before the fusion with lysosomes. From data generated through confocal laser scanning microscopy and quantification of autophagic signals we have drawn several unequivocal conclusions. The results showed a copious and rising number of autophagic organelles that had specific localization. Interestingly the increase in autophagy that occurred just after fertilization has been proved to be crucial for correct initiation of the developmental programme: irreversible developmental delays and morphologic anomalies were induced by short autophagic inhibition. This work focused on the sea urchin model system and corroborates evidence on the need for self-digestion during development, enriching the knowledge on autophagy, a biological mechanism belonging to evolutionarily different organisms.

scholarly journals Antimicrobial efficacy and biocompatibility of extracts from Cryptocarya species

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0261884
Author(s):  
Jacqueline de Oliveira Zoccolotti ◽  
Alberto José Cavalheiro ◽  
Camilla Olga Tasso ◽  
Beatriz Ribeiro Ribas ◽  
Túlio Morandin Ferrisse ◽  
...  
Keyword(s):  
High Performance ◽  
Laser Scanning ◽  
Acrylic Resin ◽  
Laser Scanning Microscopy ◽  
Live Cells ◽  
Confocal Laser ◽  
Array Detector ◽  
Control Group ◽  
Oral Keratinocytes ◽  
Denture Base

This study evaluated the efficacy of Cryptocarya spp extracts on biofilm of Candida albicans and its biocompatibility. Mature biofilm of C. albicans was formed on denture base acrylic resin samples and the fungicidal effect of the extracts was evaluated by Alamar Blue® assay, counting colony-forming units (CFU/mL) and confocal laser scanning microscopy (CLSM). Cytotoxicity of extracts from Cryptocarya species was evaluated by AlamarBlue® assay, using normal oral keratinocytes (NOK) cells. In additional, Analysis of plant extracts by ultra-high-performance liquid chromatography–diode array detector–tandem mass spectrometry (UPLC-DAD-MS) was performed. The results showed significant reduction in the cellular metabolism and in the number of CFU/mL of C. albicans (p<0.05). The concentration of 0.045 g/mL completely inhibited the number of CFU/mL. Regarding cytotoxicity, all extracts decreased cell viability compared to the control group. CLSM analysis showed predominance of live cells, but with a great difference between the groups. Antimicrobial activity of extracts from Cryptocarya on C. albicans biofilm was confirmed. However, all extracts showed toxicity on NOK cells.

scholarly journals Fluorescence Microscopy of Superplasticizers in Cementitious Systems: Applications and Challenges

Materials ◽  
2020 ◽  
Vol 13 (17) ◽  
pp. 3733
Author(s):  
Johannes Arend ◽  
Alexander Wetzel ◽  
Bernhard Middendorf
Keyword(s):  
Fluorescence Microscopy ◽  
High Performance ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Chemical System ◽  
Confocal Laser ◽  
Granulated Blast Furnace Slag ◽  
Adsorption Analysis ◽  
Resolution Imaging

In addition to the desired plasticizing effect, superplasticizers used in high and ultra-high performance concretes (UHPC) influence the chemical system of the pastes and for example retardation of the cement hydration occurs. Thus, superplasticizers have to be chosen wisely for every material composition and application. To investigate the essential adsorption of these polymers to particle surfaces in-situ to overcome several practical challenges of superplasticizer research, fluorescence microscopy is useful. In order to make the superplasticizer polymers visible for this microscopic approach, they are stained with fluorescence dyes prior the experiment. In this work, the application of this method in terms of retardation and rheological properties of sample systems is presented. The hydration of tricalcium oxy silicate (C3S) in combination with different polycarboxylate ether superplasticizers is observed by fluorescence microscopy and calorimetry. Both methods can identify the retarding effect, depending on the superplasticizer’s chemical composition. On the other hand, the influence of the superplasticizers on the slump of a ground granulated blast furnace slag/cement paste is correlated to fluorescence microscopic adsorption results. The prediction of the efficiency by microscopic adsorption analysis succeeds roughly. At last, the possibility of high-resolution imaging via confocal laser scanning microscopy is presented, which enables the detection of early hydrates and their interaction with the superplasticizers.

Effects of antimicrobial agents on oral biofilms in a saliva-conditioned flowcell

Biofilms ◽  
2004 ◽  
Vol 1 (1) ◽  
pp. 5-12 ◽  
Author(s):  
J. S. Foster ◽  
P. C. Pan ◽  
P. E. Kolenbrander
Keyword(s):  
Laser Scanning ◽  
Antimicrobial Agents ◽  
Cetylpyridinium Chloride ◽  
Oral Bacteria ◽  
Laser Scanning Microscopy ◽  
Cellular Damage ◽  
Confocal Laser ◽  
Oral Biofilms

Oral bacteria form mixed-species biofilms known as dental plaque. Growth of these complex microbial communities is often controlled with the use of antimicrobial mouthrinses. Novel laboratory methods for testing the efficacy of antimicrobials in situ are necessary to complement current clinical testing protocols. In this study, we examined the effects of antimicrobial agents on a streptococcal biofilm grown in a saliva-conditioned flowcell. The flowcell coupled with confocal laser scanning microscopy enabled examination of growing oral biofilms in situ without disruption of the microbial community. Biofilms composed of Streptococcus gordonii DL1 were grown in an in vitro flowcell and treated with several commercially available antimicrobial mouthrinses containing essential oils, triclosan, cetylpyridinium chloride/domiphen or chlorhexidine. The results of this study revealed varying abilities of the antimicrobial agents to cause cellular damage on the growing biofilm in situ. This study therefore demonstrated the usefulness of the flowcell in the rapid assessment of antimicrobial efficacy.

scholarly journals Effect of reciprocating instrumentation on chlorhexidine substantivity on human dentin: chemical analysis followed by confocal laser microscopy

2021 ◽  
Vol 37 ◽  
pp. e37038
Author(s):  
Matheus Albino Souza ◽  
Fernanda Duda Bonatto ◽  
Afonso Cristiano Fleck da Silva ◽  
Ezequiel Santin Gabrielli ◽  
Felipe Trentin Motter ◽  
...  
Keyword(s):  
High Performance ◽  
Laser Scanning ◽  
Ex Vivo ◽  
Experimental Tests ◽  
Laser Scanning Microscopy ◽  
Live Cells ◽  
Confocal Laser ◽  
Water Control ◽  
Human Teeth ◽  
Root Canals

The present research analyzed the reciprocating instrumentation associated to chlorhexidine (CHX) substantivity as its correlation with E. faecalis viability in ex vivo root canals. Eighty extracted single-rooted human teeth were used, being 40 to high-performance liquid chromatography (HPLC) and 40 to confocal laser scanning microscopy (CLSM). In both, teeth were decoronated and the cervical third was prepared. In the CLSM analysis, the root canals were inoculated with E. faecalis for 14 days. Samples were divided into 4 groups (n=10) according to instrumentation technique: no instrumentation and irrigation with distilled water (control); manual instrumentation (K-File); rotary instrumentation (ProTaper Next); and reciprocating instrumentation (Reciproc R25). Two percent chlorhexidine was applied as irrigating substance in experimental groups. Longitudinal grooves resulted in 2 halves root and 20 proof bodies in each group. Samples were divided by chance in two groups (n=10) and the outcomes were evaluated after two days and one week. The retained chlorhexidine and live cells after instrumentation techniques in each evaluation time was measured by HPLC and CLSM, respectively. Specific analysis was applied for experimental tests (p≤0.05). Both rotary as well as reciprocating techniques significantly reduced the amount of chlorhexidine on dentin in all observation periods (p<0.05). After evaluation times, all experimental groups presented lower live cells compared to control, but without statistically difference. Intragroup comparisons in times of evaluation showed no differences in instrumentation techniques, in chlorhexidine retention and number of live cells (p>0.05). Reciprocating instrumentation does not interfere on chlorhexidine substantivity.

3-D imaging of cells using a confocal laser scanning microscope and digital image processing

1988 ◽  
Vol 46 ◽  
pp. 96-97
Author(s):  
Thomas M. Jovin ◽  
Michel Robert-Nicoud ◽  
Donna J. Arndt-Jovin ◽  
Thorsten Schormann
Keyword(s):  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Scanning Microscope ◽  
Laser Scanning Microscope ◽  
Biochemical Processes ◽  
Adjacent Structures

Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.

Vacuoles Induced in Mammalian Cells by Helicobacter Cytotoxin are Acidic Organelles

1990 ◽  
Vol 48 (3) ◽  
pp. 168-169
Author(s):  
M. H. Chestnut ◽  
C. E. Catrenich
Keyword(s):  
Acridine Orange ◽  
Mammalian Cells ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ulcer Disease ◽  
Gastroduodenal Disease ◽  
H Pylori

Helicobacter pylori is a non-invasive, Gram-negative spiral bacterium first identified in 1983, and subsequently implicated in the pathogenesis of gastroduodenal disease including gastritis and peptic ulcer disease. Cytotoxic activity, manifested by intracytoplasmic vacuolation of mammalian cells in vitro, was identified in 55% of H. pylori strains examined. The vacuoles increase in number and size during extended incubation, resulting in vacuolar and cellular degeneration after 24 h to 48 h. Vacuolation of gastric epithelial cells is also observed in vivo during infection by H. pylori. A high molecular weight, heat labile protein is believed to be responsible for vacuolation and to significantly contribute to the development of gastroduodenal disease in humans. The mechanism by which the cytotoxin exerts its effect is unknown, as is the intracellular origin of the vacuolar membrane and contents. Acridine orange is a membrane-permeant weak base that initially accumulates in low-pH compartments. We have used acridine orange accumulation in conjunction with confocal laser scanning microscopy of toxin-treated cells to begin probing the nature and origin of these vacuoles.

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