Search Results (11)

In the last decade, magnetic nanoparticles (MNPs) have attracted much attention for the implementation of non-invasive approaches suitable for the diagnosis and treatment of vascular diseases. In this work, the optimization of novel vascular grafts loaded with Nickel-based nanoparticles via electrospinning is proposed. Two different polycarbonate urethanes—i.e., Corethane A80 (COT) and Chronoflex AL80 (CHF)—were used to fabricate 3D electrospun nanocomposite grafts. SEM analysis showed a homogeneous distribution of fibers, with slight differences in terms of average diameters as a function of the polymer used—(1.14 ± 0.18) µm for COT, and (1.33 ± 0.23) µm for CHF—that tend to disappear in the presence of MNPs—(1.26 ± 0.19) µm and (1.26 ± 0.213) µm for COT/NPs and CHF/NPs, respectively. TGA analyses confirmed the higher ability of CHF to entrap MNPs in the fibers—18.25% with respect to 14.63% for COT—while DSC analyses suggested an effect of MNPs on short-range rearrangements of hard/soft micro-domains of CHF. Accordingly, mechanical tests confirmed a decay of mechanical strength in the presence of MNPs with some differences depending on the matrix—from (6.16 ± 0.33) MPa to (4.55 ± 0.2) MPa (COT), and from (3.67 ± 0.18) MPa to (2.97 ± 0.22) MPa (CNF). The in vitro response revealed that the presence of MNPs did not negatively affect cell viability after 7 days in in vitro culture, suggesting a promising use of these materials as smart vascular grafts able to support the actuation function of vessel wall muscles. Full article

Carnivorous plants have fascinated botanists and ecologists with their various unusual adaptations in organ structure, physiology, and complex interactions with other organisms since the time of Charles Darwin. Species of the genus Utricularia (bladderworts, family Lentibulariaceae) are carnivorous plants that prey mainly on invertebrates using traps (bladders) of leaf origin. In the traps, there are glandular trichomes called quadrifids, which produce digestive enzymes and absorb the products of prey digestion. These quadrifids are unique due to their highly complex glandular cell structure; hence, they are an excellent model for studying the cell wall and its specialization. The main aim of the study was to investigate the presence and distribution of homogalacturonans (HGs) and hemicelluloses in the cell walls of trichome cells and especially in cell wall ingrowths in the quadrifid cells. The following antibodies were used against the wall components: anti-HGs (homogalacturonans) —JIM5 (low methylesterified HGs), JIM7 (highly esterified HGs), LM19 (low methylesterified HGs), CCRC-M38 (a fully de-esterified HG), LM5 (galactan); anti-hemicelluloses—LM25 (galactoxyloglucan; XXLLG, XXLG, XXXG modules of xyloglucans), LM15 (xyloglucan), CCRC-M138 (xylan), LM11 (heteroxylan); and anti-mannans: LM20 (heteromannan) and LM22 (heteromannan). The localization of the examined compounds was determined using immunohistochemistry techniques and immunogold labeling. In quadrifid cells, we found differences in the presence of the epitope detected by the LM5 antibody in the cell walls. In addition, cell wall ingrowths represented distinct microdomains of the cell wall in terms of the occurrence of wall components (they were methylesterified and demethylesterified homogalacturonan-poor). Hemicelluloses (galactoxyloglucan and xyloglucan) and arabinogalactans co-occur in cell wall ingrowths. Also, a part of the cell wall of the pedestal cell, which forms a Casparian strip, represented a distinct microdomain. We did not detect epitopes recognized by LM11, LM20 and LM22 antibodies. Our research shows that several cell wall microdomains occur in the cell walls of quadrifid cells. They differ depending on the presence and distribution of low methylesterified HGs, highly esterified HGs, fully de-esterified HGs, galactan (the epitope detected by the LM5 antibody), xyloglucan, galactoxyloglucan, and xylan (the epitope detected by the CCRC-M138 antibody). Full article

Since its standardization, clinical antimicrobial susceptibility testing (AST) has relied upon a standard medium, Mueller-Hinton Broth/Agar (MHB/A), to determine antibiotic resistance. However, this microbiologic medium bears little resemblance to the host milieu, calling into question the physiological relevance of resistance phenotypes it reveals. Recent studies investigating antimicrobial susceptibility in mammalian cell culture media, a more host-mimicking environment, demonstrate that exposure to host factors significantly alters susceptibility profiles. One such factor is bicarbonate, an abundant ion in the mammalian bloodstream/tissues. Importantly, bicarbonate sensitizes methicillin-resistant Staphylococcus aureus (MRSA) to early-generation β-lactams used for the treatment of methicillin-susceptible S. aureus (MSSA). This “NaHCO₃-responsive” phenotype is widespread among US MRSA USA300/CC8 bloodstream and skin and soft tissue infection isolates. Translationally, β-lactam therapy has proven effective against NaHCO₃-responsive MRSA in both ex vivo simulated endocarditis vegetation (SEV) and in vivo rabbit infective endocarditis (IE) models. Mechanistically, bicarbonate appears to influence mecA expression and PBP2a production/localization, as well as key elements for PBP2a functionality, including the PBP2a chaperone PrsA, components of functional membrane microdomains (FMMs), and wall teichoic acid (WTA) synthesis. The NaHCO₃-responsive phenotype highlights the critical role of host factors in shaping antibiotic susceptibility, emphasizing the need to incorporate more physiological conditions into AST protocols. Full article

The Utricularia (bladderworts) species are carnivorous plants that prey mainly on invertebrates using traps (bladders) of leaf origin. On the outer surfaces of the trap, there are dome-shaped glands (capitate trichomes). Each such trichome consists of a basal cell, a pedestal cell, and a terminal cell. During the maturation of these external glands, there are changes in the cell wall of the terminal cell of the gland (deposited layers of secondary wall material). Thus, due to changes in the cell wall, these glands are excellent models for studying the specialization of cell walls. The main aim of this study was to check whether different cell wall layers in terminal gland cells have a different composition in the case of homogalacturonans (low-methylesterified HGs, fully de-esterified HGs, and galactan) and hemicelluloses (galactoxyloglucan, xyloglucan, and xylan). The antibodies were used against cell wall components (anti-pectins JIM5, JIM7, LM19, CCRC-M38, and LM5 and anti-hemicelluloses LM25, LM15, CCRC-M1, and CCRC-M138). The localization of the examined compounds was determined using immunohistochemistry techniques, Carbotrace 680, and Calcofluor White. Our study showed the presence of various components in the cell walls of external gland cells: methylesterified and demethylesterified homogalacturonans, galactan, xylan, galactoxyloglucan, and xyloglucan. In the terminal cell, the primary cell wall contains different pectins in contrast to the secondary wall material, which is rich in cellulose and hemicelluloses. We also found that the basal cell differs from the other gland cells by the presence of galactan in the cell wall, which resembles the epidermal cells and parenchyma of traps. A particularly noteworthy part of the cell wall functions as a Casparian strip in the pedestal cell. Here, we found no labeling with Carbotrace 680, possibly due to cell wall modification or cell wall chemical composition variation. We have shown that the apoplastic space formed by the cell walls of the terminal cell is mainly composed of cellulose and hemicelluloses (galactoxyloglucan and xyloglucan). This composition of the cell walls allows the easy uptake of components from the external environment. Our research supports the external glands’ function as hydropotens. Full article

Species in the genus Utricularia are carnivorous plants that prey on invertebrates using traps of leaf origin. The traps are equipped with numerous different glandular trichomes. Trichomes (quadrifids) produce digestive enzymes and absorb the products of prey digestion. The main aim of this study was to determine whether arabinogalactan proteins (AGPs) occur in the cell wall ingrowths in the quadrifid cells. Antibodies (JIM8, JIM13, JIM14, MAC207, and JIM4) that act against various groups of AGPs were used. AGP localization was determined using immunohistochemistry techniques and immunogold labeling. AGPs localized with the JIM13, JIM8, and JIM14 epitopes occurred in wall ingrowths of the pedestal cell, which may be related to the fact that AGPs regulate the formation of wall ingrowths but also, due to the patterning of the cell wall structure, affect symplastic transport. The presence of AGPs in the cell wall of terminal cells may be related to the presence of wall ingrowths, but processes also involve vesicle trafficking and membrane recycling, in which these proteins participate. Full article

The genus Utricularia (bladderworts) species are carnivorous plants that prey on invertebrates using traps with a high-speed suction mechanism. The outer trap surface is lined by dome-shaped glands responsible for secreting water in active traps. In terminal cells of these glands, the outer wall is differentiated into several layers, and even cell wall ingrowths are covered by new cell wall layers. Due to changes in the cell wall, these glands are excellent models for studying the specialization of cell walls (microdomains). The main aim of this study was to check if different cell wall layers have a different composition. Antibodies against arabinogalactan proteins (AGPs) were used, including JIM8, JIM13, JIM14, MAC207, and JIM4. The localization of the examined compounds was determined using immunohistochemistry techniques and immunogold labeling. Differences in composition were found between the primary cell wall and the cell secondary wall in terminal gland cells. The outermost layer of the cell wall of the terminal cell, which was cuticularized, was devoid of AGPs (JIM8, JIM14). In contrast, the secondary cell wall in terminal cells was rich in AGPs. AGPs localized with the JIM13, JIM8, and JIM14 epitopes occurred in wall ingrowths of pedestal cells. Our research supports the hypothesis of water secretion by the external glands. Full article

Endothelial cells (ECs) respond to concurrent stimulation by biochemical factors and wall shear stress (SS) exerted by blood flow. Disruptions in flow-induced responses can result in remodeling issues and cardiovascular diseases, but the detailed mechanisms linking flow-mechanical cues and biochemical signaling remain unclear. Activin receptor-like kinase 1 (ALK1) integrates SS and ALK1-ligand cues in ECs; ALK1 mutations cause hereditary hemorrhagic telangiectasia (HHT), marked by arteriovenous malformation (AVM) development. However, the mechanistic underpinnings of ALK1 signaling modulation by fluid flow and the link to AVMs remain uncertain. We recorded EC responses under varying SS magnitudes and ALK1 ligand concentrations by assaying pSMAD1/5/9 nuclear localization using a custom multi-SS microfluidic device and a custom image analysis pipeline. We extended the previously reported synergy between SS and BMP9 to include BMP10 and BMP9/10. Moreover, we demonstrated that this synergy is effective even at extremely low SS magnitudes (0.4 dyn/cm²) and ALK1 ligand range (femtogram/mL). The synergistic response to ALK1 ligands and SS requires the kinase activity of ALK1. Moreover, ALK1’s basal activity and response to minimal ligand levels depend on endocytosis, distinct from cell–cell junctions, cytoskeleton-mediated mechanosensing, or cholesterol-enriched microdomains. However, an in-depth analysis of ALK1 receptor trafficking’s molecular mechanisms requires further investigation. Full article

Pollen tubes are tip-growing cells that create safe routes to convey sperm cells to the embryo sac for double fertilization. Recent studies have purified and biochemically characterized detergent-insoluble membranes from tobacco pollen tubes. These microdomains, called lipid rafts, are rich in sterols and sphingolipids and are involved in cell polarization in organisms evolutionarily distant, such as fungi and mammals. The presence of actin in tobacco pollen tube detergent-insoluble membranes and the preferential distribution of these domains on the apical plasma membrane encouraged us to formulate the intriguing hypothesis that sterols and sphingolipids could be a “trait d’union” between actin dynamics and polarized secretion at the tip. To unravel the role of sterols and sphingolipids in tobacco pollen tube growth, we used squalestatin and myriocin, inhibitors of sterol and sphingolipid biosynthesis, respectively, to determine whether lipid modifications affect actin fringe morphology and dynamics, leading to changes in clear zone organization and cell wall deposition, thus suggesting a role played by these lipids in successful fertilization. Full article

Carnivorous plants are unique due to their ability to attract small animals or protozoa, retain them in specialized traps, digest them, and absorb nutrients from the dissolved prey material; however, to this end, these plants need a special secretion-digestive system (glands). A common trait of the digestive glands of carnivorous plants is the presence of transfer cells. Using the aquatic carnivorous species Aldrovanda vesiculosa, we showed carnivorous plants as a model for studies of wall ingrowths/transfer cells. We addressed the following questions: Is the cell wall ingrowth composition the same between carnivorous plant glands and other plant system models? Is there a difference in the cell wall ingrowth composition between various types of gland cells (glandular versus endodermoid cells)? Fluorescence microscopy and immunogold electron microscopy were employed to localize carbohydrate epitopes associated with major cell wall polysaccharides and glycoproteins. The cell wall ingrowths were enriched with arabinogalactan proteins (AGPs) localized with the JIM8, JIM13, and JIM14 epitopes. Both methylesterified and de-esterified homogalacturonans (HGs) were absent or weakly present in the wall ingrowths in transfer cells (stalk cells and head cells of the gland). Both the cell walls and the cell wall ingrowths in the transfer cells were rich in hemicelluloses: xyloglucan (LM15) and galactoxyloglucan (LM25). There were differences in the composition between the cell wall ingrowths and the primary cell walls in A. vesiculosa secretory gland cells in the case of the absence or inaccessibility of pectins (JIM5, LM19, JIM7, LM5, LM6 epitopes); thus, the wall ingrowths are specific cell wall microdomains. Even in the same organ (gland), transfer cells may differ in the composition of the cell wall ingrowths (glandular versus endodermoid cells). We found both similarities and differences in the composition of the cell wall ingrowths between the A. vesiculosa transfer cells and transfer cells of other plant species. Full article

The ability to extrude mucilage upon seed imbibition (myxospermy) occurs in several Angiosperm taxonomic groups, but its ancestral nature or evolutionary convergence origin remains misunderstood. We investigated seed mucilage evolution in the Brassicaceae family with comparison to the knowledge accumulated in Arabidopsis thaliana. The myxospermy occurrence was evaluated in 27 Brassicaceae species. Phenotyping included mucilage secretory cell morphology and topochemistry to highlight subtle myxospermy traits. In parallel, computational biology was driven on the one hundred genes constituting the so-called A. thaliana mucilage secretory cell toolbox to confront their sequence conservation to the observed phenotypes. Mucilage secretory cells show high morphology diversity; the three studied Arabidopsis species had a specific extrusion modality compared to the other studied Brassicaceae species. Orthologous genes from the A. thaliana mucilage secretory cell toolbox were mostly found in all studied species without correlation with the occurrence of myxospermy or even more sub-cellular traits. Seed mucilage may be an ancestral feature of the Brassicaceae family. It consists of highly diverse subtle traits, probably underlined by several genes not yet characterized in A. thaliana or by species-specific genes. Therefore, A. thaliana is probably not a sufficient reference for future myxospermy evo–devo studies. Full article

Detergent-resistant membranes (DRMs) microdomains, or “raft lipids”, are key components of the plasma membrane (PM), being involved in membrane trafficking, signal transduction, cell wall metabolism or endocytosis. Proteins imbibed in these domains play important roles in these cellular functions, but there are few studies concerning DRMs under abiotic stress. In this work, we determine DRMs from the PM of broccoli roots, the lipid and protein content, the vesicles structure, their water osmotic permeability and a proteomic characterization focused mainly in aquaporin isoforms under salinity (80 mM NaCl). Based on biochemical lipid composition, higher fatty acid saturation and enriched sterol content under stress resulted in membranes, which decreased osmotic water permeability with regard to other PM vesicles, but this permeability was maintained under control and saline conditions; this maintenance may be related to a lower amount of total PIP1 and PIP2. Selective aquaporin isoforms related to the stress response such as PIP1;2 and PIP2;7 were found in DRMs and this protein partitioning may act as a mechanism to regulate aquaporins involved in the response to salt stress. Other proteins related to protein synthesis, metabolism and energy were identified in DRMs independently of the treatment, indicating their preference to organize in DMRs. Full article