Novel tool to quantify cell wall porosity relates wall structure to cell growth and drug uptake

Even though cell walls have essential functions for bacteria, fungi, and plants, tools to investigate their dynamic structure in living cells have been missing. Here, it is shown that changes in the intensity of the plasma membrane dye FM4-64 in response to extracellular quenchers depend on the nano-scale porosity of cell walls. The correlation of quenching efficiency and cell wall porosity is supported by tests on various cell types, application of differently sized quenchers, and comparison of results with confocal, electron, and atomic force microscopy images. The quenching assay was used to investigate how changes in cell wall porosity affect the capability for extension growth in the model plant Arabidopsis thaliana. Results suggest that increased porosity is not a precondition but a result of cell extension, thereby providing new insight on the mechanism plant organ growth. Furthermore, it was shown that higher cell wall porosity can facilitate the action of antifungal drugs in Saccharomyces cerevisiae, presumably by facilitating uptake.

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Susceptibility Pattern of Caspofungin-Coated Gold Nanoparticles against Clinically Important Candida Species

Advanced Pharmaceutical Bulletin ◽

10.34172/apb.2021.078 ◽

2020 ◽

Author(s):

Zahra Salehi ◽

Azam Fattahi ◽

Ensieh lotfali ◽

Abdolhassan Kazemi ◽

Ali Shakeri-Zadeh ◽

...

Keyword(s):

Electron Microscopy ◽

Gold Nanoparticles ◽

Cell Wall ◽

Candida Species ◽

Spherical Shape ◽

Sem Analysis ◽

Antifungal Drugs ◽

Wall Structure ◽

Candida Spp ◽

Average Size

Purpose: The present study was performed to examine whether caspofungin-coated gold nanoparticles (CAS-AuNPs) may offer the right platform for sensitivity induction in resistant isolates. Methods: For the purpose of the study, a total of 58 archived Candida species were enrolled in the research. The identification of Candida spp. was performed using polymerase chain reaction-restriction fragment length polymorphism and HWP1 gene amplification approaches. The conjugated CAS-AuNPs were synthesized and then characterized using transmission electron microscopy (TEM) and Zetasizer system to determine their morphology, size, and charge. Furthermore, the efficacy of CAS, CAS-AuNPs conjugate, and AuNPs against Candida spp. was assessed based on the Clinical and Laboratory Standards Institute M60. Finally, the interaction of CAS-AuNPs with Candida element was evaluated via scanning electron microscopy (SEM). Results: According to the TEM results, the synthesized CAS-AuNPs had a spherical shape with an average size of 20 nm. The Zeta potential of CAS-AuNPs was -38.2 mV. Statistical analyses showed that CAS-AuNPs could significantly reduce the minimum inhibitory concentration against C. albicans (P=0.0005) and non-albicans Candida (NAC) species (P<0.0001). All isolates had a MIC value of ≥ 4 µg/ml for CAS, except for C. glabrata. The results of SEM analysis confirmed the effects of AuNPs on the membrane and cell wall structure of C. globrata exposed to CAS-AuNPs, facilitating the formation of pores on the cell wall and finally cell death. Conclusion: The findings revealed that CAS-AuNPs conjugates had significant antifungal effects against Candida spp. through the degradation of the membrane and cell wall integrity. Therefore, it can be concluded that the encapsulation of antifungal drugs in combination with NPs not only diminishes side effects but also enhances the effectiveness of the medications.

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Evaluation of Antifungal Activity and Mode of Action of New Coumarin Derivative, 7-Hydroxy-6-nitro-2H-1-benzopyran-2-one, againstAspergillusspp.

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2015/925096 ◽

2015 ◽

Vol 2015 ◽

pp. 1-8 ◽

Cited By ~ 11

Author(s):

Felipe Queiroga Sarmento Guerra ◽

Rodrigo Santos Aquino de Araújo ◽

Janiere Pereira de Sousa ◽

Fillipe de Oliveira Pereira ◽

Francisco J. B. Mendonça-Junior ◽

...

Keyword(s):

Cell Wall ◽

Mode Of Action ◽

Cell Walls ◽

Coumarin Derivative ◽

Antifungal Drugs ◽

Azole Resistance ◽

Subinhibitory Concentration ◽

Fungal Cell ◽

Mycelia Growth

Aspergillusspp. produce a wide variety of diseases. For the treatment of such infections, the azoles and Amphotericin B are used in various formulations. The treatment of fungal diseases is often ineffective, because of increases in azole resistance and their several associated adverse effects. To overcome these problems, natural products and their derivatives are interesting alternatives. The aim of this study was to examine the effects of coumarin derivative, 7-hydroxy-6-nitro-2H-1-benzopyran-2-one (Cou-NO2), both alone and with antifungal drugs. Its mode of action againstAspergillusspp. Cou-NO2was tested to evaluate its effects on mycelia growth and germination of fungal conidia ofAspergillusspp. We also investigated possible Cou-NO2action on cell walls (0.8 M sorbitol) and on Cou-NO2to ergosterol binding in the cell membrane. The study shows that Cou-NO2is capable of inhibiting both the mycelia growth and germination of conidia for the species tested, and that its action affects the structure of the fungal cell wall. At subinhibitory concentration, Cou-NO2enhanced thein vitroeffects of azoles. Moreover, in combination with azoles (voriconazole and itraconazole) Cou-NO2displays an additive effect. Thus, our study supports the use of coumarin derivative 7-hydroxy-6-nitro-2H-1-benzopyran-2-one as an antifungal agent againstAspergillusspecies.

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Investigation of the Mechanical Properties of Flax Cell Walls during Plant Development: The Relation between Performance and Cell Wall Structure

Fibers ◽

10.3390/fib6010006 ◽

2018 ◽

Vol 6 (1) ◽

pp. 6 ◽

Cited By ~ 17

Author(s):

Camille Goudenhooft ◽

David Siniscalco ◽

Olivier Arnould ◽

Alain Bourmaud ◽

Olivier Sire ◽

...

Keyword(s):

Mechanical Properties ◽

Cell Wall ◽

Plant Development ◽

Cell Walls ◽

Cell Wall Structure ◽

Wall Structure

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Up against the Wall: Is Yeast Cell Wall Integrity Ensured by Mechanosensing in Plasma Membrane Microdomains?

Applied and Environmental Microbiology ◽

10.1128/aem.03273-14 ◽

2014 ◽

Vol 81 (3) ◽

pp. 806-811 ◽

Cited By ~ 42

Author(s):

Christian Kock ◽

Yves F. Dufrêne ◽

Jürgen J. Heinisch

Keyword(s):

Cell Wall ◽

Plasma Membrane ◽

Yeast Cell ◽

Antifungal Drugs ◽

Cell Wall Integrity ◽

Yeast Cell Wall ◽

Membrane Microdomains ◽

Wall Structure ◽

Fungal Cell ◽

Membrane Spanning

ABSTRACTYeast cell wall integrity (CWI) signaling serves as a model of the regulation of fungal cell wall synthesis and provides the basis for the development of antifungal drugs. A set of five membrane-spanning sensors (Wsc1 to Wsc3, Mid2, and Mtl1) detect cell surface stress and commence the signaling pathway upon perturbations of either the cell wall structure or the plasma membrane. We here summarize the latest advances in the structure/function relationship primarily of the Wsc1 sensor and critically review the evidence that it acts as a mechanosensor. The relevance and physiological significance of the information obtained for the function of the other CWI sensors, as well as expected future developments, are discussed.

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The fate of acetyl groups and sugar components during the digestion of grass cell walls in sheep

The Journal of Agricultural Science ◽

10.1017/s0021859600028252 ◽

1977 ◽

Vol 89 (2) ◽

pp. 327-340 ◽

Cited By ~ 55

Author(s):

E. Jane Morris ◽

J. S. D. Bacon

Keyword(s):

Cell Wall ◽

Cell Walls ◽

Lignin Content ◽

Sodium Ethoxide ◽

Mesophyll Cell ◽

Cell Types ◽

Gas Liquid Chromatography ◽

Cell Wall Preparation ◽

The Relationship ◽

Isolated Cell

SummaryThe digestibilities of grass cell wall constituents determined in a digestion trial were compared with those obtained by suspending various isolated cell wall preparations in nylon bags in the rumen of a sheep. Particular attention was paid to acetyl groups and to individual sugars, which were determined in both cases by gas liquid chromatography.For dried grass and hay in the digestion trial the cell wall constituents showed digestibilities decreasing in the following order: arabinose, galactose, glucose, xylose, acetyl, lignin.For a leaf cell wall preparation derived from all cell types except mesophyll, the nylon bag technique allowed the same order of digestibilities; rhamnose and uronic acids were also measured and found to be rapidly digested. Mesophyll cell walls placed in nylon bags were more readily digested than non-mesophyll. All the sugars, and also acetyl groups, were digested to the same extent.In a grass cell wall preparation isolated from sheep faeces, tested similarly, xylose and glucose were digested to the same extent, but acetyl groups were less digested.Removal of acetyl groups, using sodium ethoxide, which left the sugar composition and lignin content unchanged, increased the digestibility particularly of the cell walls from faeces.The results are discussed with reference to the relationship between cell wall composition and digestibility.

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Die gemeinsame Wurzel der Lyse von Escherichia coli durch Penicillin oder durch Phagen

Zeitschrift für Naturforschung B ◽

10.1515/znb-1957-0701 ◽

1957 ◽

Vol 12 (7) ◽

pp. 421-427 ◽

Cited By ~ 51

Author(s):

W. Weidel ◽

J. Primosigh

Keyword(s):

Cell Wall ◽

Cell Walls ◽

Building Blocks ◽

Diaminopimelic Acid ◽

Wall Structure ◽

Gram Positive ◽

E Coli ◽

Growing Cell ◽

Cell Wall Disruption ◽

Entire Cell

One of the two layers of the E. coli B cell wall is shown to possess the chemical composition typical of a gram-positive microorganism. It is this layer which lends support and strength to the entire cell wall structure, its rigidity depending up on the incorporation of building blocks made up from alanine, glutamic acid, diaminopimelic acid, muramic acid and glucosamine.Phage enzyme is an agent capable of removing these stabilizing units from the „gram-positive “ layer, thereby causing it to collapse. Penicillin appears to prevent the biosynthetic incorporation of the same stabilizing units into growing cell walls, thus producing eventually the effect of cell wall disruption in a basically similar way.The rather manifold aspects of these findings are discussed at some length.

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Chilling Injury in Peaches: A Cytochemical and Ultrastructural Cell Wall Study

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.117.1.114 ◽

1992 ◽

Vol 117 (1) ◽

pp. 114-118 ◽

Cited By ~ 53

Author(s):

J.G. Luza ◽

R. van Gorsel ◽

V.S. Polito ◽

A.A. Kader

Keyword(s):

Cell Wall ◽

Cell Walls ◽

Prunus Persica ◽

Periodic Acid ◽

Chilling Injury ◽

Middle Lamella ◽

Wall Structure ◽

Positive Staining ◽

Intercellular Matrix ◽

Parenchyma Cells

Fruits of mid- (`O'Henry'), late (`Airtime'), and extra-late-season (`Autumn Gem') peach [Prunus persica (L.) Batsch] cultivars were examined for changes in cell wall structure and cytochemistry that accompany the onset of mealiness and leatheriness of the mesocarp due to chilling injury. The peaches were stored at 10C for up to 18 days or at SC for up to 29 days. Plastic-embedded sections were stained by the Schiff's-periodic acid reaction, Calcofluor white MR2, and Coriphosphine to demonstrate total insoluble carbohydrates, ß-1,4 glucans, and pectins, respectively. Mealiness was characterized by separation of mesocarp parenchyma cells leading to increased intercellular spaces and accumulation of pectic substances in the intercellular matrix. Little structural change was apparent in the cellulosic component of the cell walls of these fruits. In leathery peaches, the mesocarp parenchyma cells collapsed, intercellular space continued to increase, and pectin-positive staining in the intercellular matrix increased greatly. In addition, the component of the cell walls that stained positively for ß-1,4 glucans became thickened relative to freshly harvested or mealy fruit. At the ultrastructural level, dissolution of the middle lamella, cell separation, irregular thickening of the primary wall, and plasmolysis of the mesocarp parenchyma cells were seen as internal breakdown progressed.

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Aplikasi Kalsium dan NAA untuk Mengendalikan Getah Kuning Buah Manggis (Garcinia mangostana L.)

Jurnal Hortikultura Indonesia ◽

10.29244/jhi.9.1.10-18 ◽

2018 ◽

Vol 9 (1) ◽

pp. 10-18

Author(s):

Yulinda Tanari ◽

Darda Efendi ◽

Roedhy Poerwanto ◽

Didy Sopandie ◽

Ketty Suketi

Keyword(s):

Cell Wall ◽

Cell Division ◽

Cell Walls ◽

Block Design ◽

Wall Structure ◽

Sink Strength ◽

Naphthaleneacetic Acid ◽

Garcinia Mangostana ◽

Random Block ◽

Random Block Design

ABSTRACTThe yellow sap is produced naturally in mangosteen organ except in the root. The yellow sap contaminated the aryl and rind if the epithelial cell walls rupture due to deficiency of calcium (Ca). Calcium is one of structural component of cell walls, whereas naphthaleneacetic acid (NAA) has its role in improving cell division and cell elongation. Interaction of Ca and NAA can improve sink strength and capacity because the newly formed cells need Ca to construct wall structure. This experiment aimed at finding out the effect of Ca and NAA applications in reducing the contamination of yellow sap in mangosteen. The experiment was conducted by using factorial random block design consisting of 2 factors and 3 replications. The first factor was Ca dosage (0 and 4.8 kg/tree), and the second factor was NAA concentration (0, 200, 400 and 600 ppm). The results showed that application of 4.8 Ca/tree and 200 ppm NAA as much as 5 ml / fruit effectively improve the content of Ca pectate in pericarp, reduced the percentage of yellow sap contamination on the fruit segment, aryl and rind to 0% and 12.3% respectively compared to control (17.8% on fruit segment, 36.8% on aryl and 56.1% on rind).Key words: aryl, Ca pectate, cell wall, middle lamela.ABSTRAKGetah kuning adalah getah yang dihasilkan secara alami pada setiap organ manggis, kecuali pada akar. Getah kuning akan keluar dan mencemari aril serta kulit jika dinding sel epitel pecah karena kekurangan kalsium (Ca). Kalsium adalah komponen dinding sel, berperan dalam struktur dan permeabilitas membran sedangkan asam naftalenasetat (NAA) berperan penting dalam meningkatkan pembelahan dan pembesaran sel. Interaksi keduanya dapat meningkatkan kapasitas sink buah karena sel yang baru terbentuk membutuhkan Ca dalam menyusun struktur dinding sel. Percobaan bertujuan untuk mengetahui pengaruh aplikasi Ca dan NAA dalam menurunkan cemaran getah kuning manggis. Percobaan menggunakan rancangan acak kelompok faktorial 2 faktor dengan 3 ulangan. Faktor ke-1 yaitu dosis Ca (0 dan 4.8 kg Ca/pohon) dan faktor ke-2 yaitu konsentrasi NAA (0, 200, 400 dan 600 ppm) dengan volume semprot 5 ml perbuah. Hasil percobaan menunjukkan bahwa aplikasi 4.8 kg Ca/pohon dengan NAA 200 ppm sebanyak 5 ml/buah efektif meningkatkan kandungan Ca pektat perikarp dan menurunkan persentase buah tercemar getah kuning menjadi 0% pada juring dan aril serta 12.3% pada kulit dibandingkan dengan perlakuan kontrol (17.8% pada juring, 36.8% pada aril dan 56.1% pada kulit buah).Kata kunci: aril, Ca pektat, dinding sel, lamela tengah

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Staphylococcus aureus cell wall structure and dynamics during host-pathogen interaction

PLoS Pathogens ◽

10.1371/journal.ppat.1009468 ◽

2021 ◽

Vol 17 (3) ◽

pp. e1009468

Author(s):

Joshua A. F. Sutton ◽

Oliver T. Carnell ◽

Lucia Lafage ◽

Joe Gray ◽

Jacob Biboy ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Cell Wall ◽

Cell Walls ◽

Ex Vivo ◽

Cell Wall Structure ◽

Wall Structure ◽

Bacterial Cells ◽

Human Macrophages ◽

Structure And Dynamics

Peptidoglycan is the major structural component of the Staphylococcus aureus cell wall, in which it maintains cellular integrity, is the interface with the host, and its synthesis is targeted by some of the most crucial antibiotics developed. Despite this importance, and the wealth of data from in vitro studies, we do not understand the structure and dynamics of peptidoglycan during infection. In this study we have developed methods to harvest bacteria from an active infection in order to purify cell walls for biochemical analysis ex vivo. Isolated ex vivo bacterial cells are smaller than those actively growing in vitro, with thickened cell walls and reduced peptidoglycan crosslinking, similar to that of stationary phase cells. These features suggested a role for specific peptidoglycan homeostatic mechanisms in disease. As S. aureus penicillin binding protein 4 (PBP4) has reduced peptidoglycan crosslinking in vitro its role during infection was established. Loss of PBP4 resulted in an increased recovery of S. aureus from the livers of infected mice, which correlated with enhanced fitness within murine and human macrophages. Thicker cell walls correlate with reduced activity of peptidoglycan hydrolases. S. aureus has a family of 4 putative glucosaminidases, that are collectively crucial for growth. Loss of the major enzyme SagB, led to attenuation during murine infection and reduced survival in human macrophages. However, loss of the other three enzymes Atl, SagA and ScaH resulted in clustering dependent attenuation, in a zebrafish embryo, but not a murine, model of infection. A combination of pbp4 and sagB deficiencies resulted in a restoration of parental virulence. Our results, demonstrate the importance of appropriate cell wall structure and dynamics during pathogenesis, providing new insight to the mechanisms of disease.

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Bundling of cellulose microfibrils in native and polyethylene glycol-containing wood cell walls revealed by small-angle neutron scattering

Scientific Reports ◽

10.1038/s41598-020-77755-y ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Paavo A. Penttilä ◽

Michael Altgen ◽

Muhammad Awais ◽

Monika Österberg ◽

Lauri Rautkari ◽

...

Keyword(s):

Cell Wall ◽

Neutron Scattering ◽

Plant Cell ◽

Small Angle ◽

Cell Walls ◽

Small Angle Neutron Scattering ◽

Plant Cell Wall ◽

Raw Materials ◽

Cellulose Microfibrils ◽

Wall Structure

AbstractWood and other plant-based resources provide abundant, renewable raw materials for a variety of applications. Nevertheless, their utilization would greatly benefit from more efficient and accurate methods to characterize the detailed nanoscale architecture of plant cell walls. Non-invasive techniques such as neutron and X-ray scattering hold a promise for elucidating the hierarchical cell wall structure and any changes in its morphology, but their use is hindered by challenges in interpreting the experimental data. We used small-angle neutron scattering in combination with contrast variation by poly(ethylene glycol) (PEG) to identify the scattering contribution from cellulose microfibril bundles in native wood cell walls. Using this method, mean diameters for the microfibril bundles from 12 to 19 nm were determined, without the necessity of cutting, drying or freezing the cell wall. The packing distance of the individual microfibrils inside the bundles can be obtained from the same data. This finding opens up possibilities for further utilization of small-angle scattering in characterizing the plant cell wall nanostructure and its response to chemical, physical and biological modifications or even in situ treatments. Moreover, our results give new insights into the interaction between PEG and the wood nanostructure, which may be helpful for preservation of archaeological woods.

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