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International Journal of Molecular... Jul 2020Although cell wall dynamics, particularly modification of homogalacturonan (HGA, a major component of pectin) during pollen tube growth, have been extensively studied in...
Although cell wall dynamics, particularly modification of homogalacturonan (HGA, a major component of pectin) during pollen tube growth, have been extensively studied in dicot plants, little is known about how modification of the pollen tube cell wall regulates growth in monocot plants. In this study, we assessed the role of HGA modification during elongation of the rice pollen tube by adding a pectin methylesterase (PME) enzyme or a PME-inhibiting catechin extract (Polyphenon 60) to in vitro germination medium. Both treatments led to a severe decrease in the pollen germination rate and elongation. Furthermore, using monoclonal antibodies toward methyl-esterified and de-esterified HGA epitopes, it was found that exogenous treatment of PME and Polyphenon 60 resulted in the disruption of the distribution patterns of low- and high-methylesterified pectins upon pollen germination and during pollen tube elongation. Eleven PMEs and 13 PME inhibitors (PMEIs) were identified by publicly available transcriptome datasets and their specific expression was validated by qRT-PCR. Enzyme activity assays and subcellular localization using a heterologous expression system in tobacco leaves demonstrated that some of the pollen-specific PMEs and PMEIs possessed distinct enzymatic activities and targeted either the cell wall or other compartments. Taken together, our findings are the first line of evidence showing the essentiality of HGA methyl-esterification status during the germination and elongation of pollen tubes in rice, which is primarily governed by the fine-tuning of PME and PMEI activities.
Topics: Carboxylic Ester Hydrolases; Cell Wall; Gene Expression Regulation, Plant; Germination; Oryza; Pectins; Plant Leaves; Plant Proteins; Pollen Tube; Polyphenols; Nicotiana; Transcriptome
PubMed: 32650624
DOI: 10.3390/ijms21144840 -
International Journal of Biological... Oct 2019Several studies have been conducted on polysaccharides derived from natural sources, and their different biological properties have been reported. Their low toxicity and...
Several studies have been conducted on polysaccharides derived from natural sources, and their different biological properties have been reported. Their low toxicity and antiviral effects i.e., their action on several steps of viral replication, have been extensively examined. In this work, pectin isolated from Inga spp. fruit pulp was first characterized and evaluated using HEp-2 cells against the herpes simplex virus type 1 (HSV-1) and the poliovirus (PV). The isolated pectin (denoted as PDTS) was characterized by infrared spectroscopy, NMR and Gel permeation chromatography. The cytotoxicity was analyzed by the MTT method and antiviral activity by plaque reduction assay, immunofluorescence assay (IF) and polymerase chain reaction (PCR). The cytotoxic concentration (CC) of PDTS was 870 μg.mL and the inhibitory concentrations (IC) were 179 μg.mL and 58 μg.mL for HSV-1 and PV, respectively. Greater inhibitory effect was observed when the cells were simultaneously treated with PDTS and infected, suggesting that PDTS inhibited the initial viral replication stages, revealing its antiviral potential.
Topics: Antiviral Agents; Cell Line; Fabaceae; Herpesvirus 1, Human; Pectins; Poliovirus; Virus Replication
PubMed: 31376449
DOI: 10.1016/j.ijbiomac.2019.07.212 -
Pharmaceutics May 2024A general procedure to prepare gold nanourchins (GNUs) via a seed-mediated method was followed using dopamine hydrochloride as a reducing agent and silver nitrate salt...
A general procedure to prepare gold nanourchins (GNUs) via a seed-mediated method was followed using dopamine hydrochloride as a reducing agent and silver nitrate salt (AgNO) as a shape-directing agent. The novelty of this study comes from the successful incorporation of the prepared gold urchins as an aqueous suspension in a nasal pressurized metered dose inhaler (pMDI) formulation and the investigation of their potential for olfactory targeting for direct nose-to-brain drug delivery (NTBDD). The developed pMDI formulation was composed of 0.025% / GNUs, 2% / Milli-Q water, and 2% / EtOH, with the balance of the formulation being HFA134a propellant. Particle integrity and aerosolization performance were examined using an aerosol exposure system, whereas the nasal deposition profile was tested in a sectioned anatomical replica of human nasal airways. The compatibility of the gold dispersion with the nasal epithelial cell line RPMI 2650 was also investigated in this study. Colloidal gold was found to be stable following six-month storage at 4 °C and during the lyophilization process utilizing a pectin matrix for complete re-dispersibility in water. The GNUs were intact and discrete following atomization via a pMDI, and 13% of the delivered particles were detected beyond the nasal valve, the narrowest region in the nasal cavity, out of which 5.6% was recovered from the olfactory region. Moreover, the formulation was found to be compatible with the human nasal epithelium cell line RPMI 2650 and excellent cell viability was observed. The formulated GNU-HFA-based pMDI is a promising approach for intranasal drug delivery, including deposition in the olfactory region, which could be employed for NTBDD applications.
PubMed: 38794331
DOI: 10.3390/pharmaceutics16050669 -
Pharmaceutics Aug 2023The aim of this project is to use pectin- and chitosan-modified solid lipid nanoparticles for bovine lactoferrin to enhance its cellular uptake and transport.
AIM
The aim of this project is to use pectin- and chitosan-modified solid lipid nanoparticles for bovine lactoferrin to enhance its cellular uptake and transport.
METHODS
Solid lipid particles containing bovine lactoferrin (bLf) were formulated through the solvent evaporation technique, incorporating stearic acid along with either chitosan or pectin modification. bLf cellular uptake and transport were evaluated in vitro using the human adenocarcinoma cell line Caco-2 cell model.
RESULTS AND DISCUSSION
The bLf-loaded SLPs showed no significant effect on cytotoxicity and did not induce apoptosis within the eight-hour investigation. The use of confocal laser scanning microscopy confirmed that bLf follows the receptor-mediated endocytosis, whereas the primary mechanism for the cellular uptake of SLPs was endocytosis. The bLf-loaded SLPs had significantly more cellular uptake compared to bLf alone, and it was observed that this impact varied based on the time, temperature, and concentration. Verapamil and EDTA were determined to raise the apparent permeability coefficients (App) of bLf and bLf-loaded SLPs.
CONCLUSION
This occurred because they hindered efflux by interacting with P-glycoproteins and had a penetration-enhancing influence. These findings propose the possibility of an additional absorption mechanism for SLPs, potentially involving active transportation facilitated by the P-glycoprotein transporter in Caco-2 cells. These results suggest that SLPs have the potential to be applied as effective carriers to improve the oral bioavailability of proteins and peptides.
PubMed: 37631382
DOI: 10.3390/pharmaceutics15082168 -
Heliyon Apr 2023Diabetes remains a global public health threat because of its increasing prevalence and mortality, especially in people under the age of 25. Metformin hydrochloride...
Diabetes remains a global public health threat because of its increasing prevalence and mortality, especially in people under the age of 25. Metformin hydrochloride (HCl), as recommended by American Diabetes Association in 2022, is the first-line therapy for type 2 diabetes in adults. Metformin has low oral bioavailability due to poor permeability. Therefore, by developing metformin HCl oral in situ gel, sustained delivery of metformin can be achieved, thus enhancing the absorption of the drug. Sodium alginate and pectin were used for formulating the system. Different adjuvant polymers, including HPMC K4M, HPMC K100 LV, PEG 4000, and SCMC were used as released-pattern-modifying agents. All formulations could afloat in 0.1 N HCl at the pH of 1.2 within a minute and stay afloat for over 8 h. The optimized formulation could be made from either sodium alginate (2%) and HPMC K4M (0.5%) or pectin (2%) and HPMC K4M (2%). The optimized formulations gradually released metformin HCl with a cumulative release of 80% within 8 h. We successfully developed floating in situ gels that can release metformin HCl sustainedly.
PubMed: 37025765
DOI: 10.1016/j.heliyon.2023.e14796 -
ACS Applied Bio Materials Aug 2023Natural polymers from organic wastes have gained increasing attention in the biomedical field as resourceful second raw materials for the design of biomedical devices...
Natural polymers from organic wastes have gained increasing attention in the biomedical field as resourceful second raw materials for the design of biomedical devices which can perform a specific bioactive function and eventually degrade without liberating toxic residues in the surroundings. In this context, patches and bandages, that need to support the skin wound healing process for a short amount of time to be then discarded, certainly constitute good candidates in our quest for a more environmentally friendly management. Here, we propose a plant-based microfibrous scaffold, loaded with vitamin C (VitC), a bioactive molecule which acts as a protecting agent against UV damages and as a wound healing promoter. Fibers were fabricated via electrospinning from various zein/pectin formulations, and subsequently cross-linked in the presence of Ca to confer them a hydrogel-like behavior, which we exploited to tune both the drug release profile and the scaffold degradation. A comprehensive characterization of the physico-chemical properties of the zein/pectin/VitC scaffolds, either pristine or cross-linked, has been carried out, together with the bioactivity assessment with two representative skin cell populations (human dermal fibroblast cells and skin keratinocytes, HaCaT cells). Interestingly, col-1a gene expression of dermal fibroblasts increased after 3 days of growth in the presence of the microfiber extraction media, indicating that the released VitC was able to stimulate collagen mRNA production overtime. Antioxidant activity was analyzed on HaCaT cells via DCFH-DA assay, highlighting a fluorescence intensity decrease proportional to the amount of loaded VitC (down to 50 and 30%), confirming the protective effect of the matrices against oxidative stress. Finally, the most performing samples were selected for the in vivo test on a skin UVB-burn mouse model, where our constructs demonstrated to significantly reduce the inflammatory cytokines expression in the injured area (50% lower than the control), thus constituting a promising, environmentally sustainable alternative to skin patches.
Topics: Animals; Humans; Male; Mice; Antioxidants; Ascorbic Acid; Biocompatible Materials; Burns; Cell Line; Hydrogels; Keratinocytes; Mice, Inbred C57BL; Wound Healing; Zein; Zea mays
PubMed: 37493659
DOI: 10.1021/acsabm.3c00214 -
Cells Mar 2020Softening of fruit during the postharvest storage, which is mainly associated with both compositional and spatial changes of polysaccharides within cell wall, affects...
Hybrid Label-Free Molecular Microscopies for Simultaneous Visualization of Changes in Cell Wall Polysaccharides of Peach at Single- and Multiple-Cell Levels during Postharvest Storage.
Softening of fruit during the postharvest storage, which is mainly associated with both compositional and spatial changes of polysaccharides within cell wall, affects the texture and quality of fruit. Current research on the fruit softening mechanism lacks an understanding of the overall softening at the cell level. The objective of this work was to investigate the change in the spatial distribution of cell wall polysaccharides in peach flesh cells at both single- and multiple-cell levels in a label-free way during the postharvest storage. Nonmelting peaches ( L. Batsch cv."Zhonghuashoutao") at commercial maturity were stored at 0 °C and 20 °C. Firmness measurement and chemical analysis were performed at each storage time. In addition, three molecular imaging techniques, namely confocal Raman microspectroscopy (CRM), Fourier transform infrared microspectroscopy (FTIRM), and stimulated Raman scattering microscopy (SRS) were used to visualize changes in the spatial distribution of cell wall polysaccharides of peach fruit in a label-free way during the postharvest storage. The combination of CRM and FTIRM provided complementary spectral information to visualize the spatial changes of cellulose, hemicellulose, and pectin in the cell wall of peach flesh during softening at the single-cell level, and found that the cell wall polysaccharides tended to be concentrated in the cell corner of parenchymal cells at the late stage. Furthermore, SRS, which is an ultrafast Raman imaging technique (approximately three or four orders of magnitude faster than CRM), was used for high-throughput cell wall phenotypes measurement. Different degradation degrees of parenchymal cells during fruit softening were found based on the gray-scale statistical analysis of SRS data. In general, cell wall polysaccharides decreased during softening and tended to be concentrated in the cell corner for most parenchymal cells at the late stage, but there were also some cells not in line with the whole softening trends. The results show that there were differences in the content and spatial changes of cell wall polysaccharides among parenchymal cells of peach fruit during the softening process, and the hybrid use of CRM, FTIRM, and SRS is a promising method for simultaneous visualization of changes in cell wall polysaccharides of peach.
Topics: Cell Wall; Hyperspectral Imaging; Microscopy, Confocal; Polysaccharides; Preservation, Biological; Prunus persica; Single-Cell Analysis; Spectroscopy, Fourier Transform Infrared; Spectrum Analysis, Raman
PubMed: 32244921
DOI: 10.3390/cells9030761 -
Gastrointestinal Endoscopy Mar 2024Traction has become the reference strategy for colorectal endoscopic submucosal dissection (ESD). One of its major limitations is that the force of traction decreases as...
BACKGROUND AND AIMS
Traction has become the reference strategy for colorectal endoscopic submucosal dissection (ESD). One of its major limitations is that the force of traction decreases as dissection progresses. The ATRACT traction device uses a pulley system to increase traction during the procedure, making it easier and faster. A retrospective study of 54 cases showed interesting results in terms of efficacy and safety throughout the digestive tract. We sought to confirm these initial results with a prospective study of resections of colorectal lesions.
METHODS
In this prospective multicenter study, 5 experienced operators from 3 different centers each performed 10 procedures using the ATRACT device consecutively for all conventional colorectal ESDs measuring between 4 and 10 cm in conventional locations (no recurrent lesions, appendicular, of the ileo-cecal valve, in contact with the pectinate line or measuring more than 2/3 of the circumference).
RESULTS
Between November 2022 and April 2023, 50 ESDs were performed in 49 patients. On average, the main diameter of the lesions was 66.6 mm, with a surface area of 3066 mm2. The mean operating time was 55.2 minutes, resulting in a mean resection speed of 61.4 mm2/min. En Bloc and R0 resection rates were 100% and 98% respectively. Resections were curative in 94% of cases. 4 perforations (8%) occurred, all of which were closed endoscopically without the need for surgery. 1 case of delayed hemorrhage (2%) was noted. To date, this is the series with the highest resection speeds in the literature for colorectal ESD. For comparison, Yamamoto et al. reported a speed of 23.5 mm2/min using the "pocket" strategy and Bordillon et al. reported a speed of 39.1 mm2/min using the double-clip traction technique. These results need to be confirmed in larger studies, and in non-expert centers.
CONCLUSIONS
This prospective evaluation confirms the efficacy and safety of the adaptive traction strategy with the ATRACT device for colorectal ESD with high resection speed.
PubMed: 38458261
DOI: 10.1016/j.gie.2024.02.032 -
Carbohydrate Polymers Jan 2021Pectins have anti-inflammatory effects via Toll-like receptor (TLR) inhibition in a degree of methyl-esterification-(DM)-dependent manner. However, pectins also vary in...
Pectins have anti-inflammatory effects via Toll-like receptor (TLR) inhibition in a degree of methyl-esterification-(DM)-dependent manner. However, pectins also vary in distribution of methyl-esters over the galacturonic-acid (GalA) backbone (Degree of Blockiness - DB) and impact of this on anti-inflammatory capacity is unknown. Pectins mainly inhibit TLR2-1 but magnitude depends on both DM and DB. Low DM pectins (DM18/19) with both low (DB86) and high DB (DB94) strongly inhibit TLR2-1. However, pectins with intermediate DM (DM43/DM49) and high DB (DB60), but not with low DB (DB33), inhibit TLR2-1 as strongly as low DM. High DM pectins (DM84/88) with DB71 and DB91 do not inhibit TLR2-1 strongly. Pectin-binding to TLR2 was confirmed by capture-ELISA. In human macrophages, low DM and intermediate DM pectins with high DB inhibited TLR2-1 induced IL-6 secretion. Both high number and blockwise distribution of non-esterified GalA in pectins are responsible for the anti-inflammatory effects via inhibition of TLR2-1.
Topics: Anti-Inflammatory Agents; Cell Line; Chromatography, High Pressure Liquid; Esterification; Esters; Hexuronic Acids; Humans; Inflammation; Macrophages; Pectins; Toll-Like Receptor 2
PubMed: 33152851
DOI: 10.1016/j.carbpol.2020.117093 -
Frontiers in Plant Science 2021Gall formation on the belowground parts of plants infected with is the result of extensive host cellular reprogramming. The development of these structures is a...
Gall formation on the belowground parts of plants infected with is the result of extensive host cellular reprogramming. The development of these structures is a consequence of increased cell proliferation followed by massive enlargement of cells colonized with the pathogen. Drastic changes in cellular growth patterns create local deformities in the roots and hypocotyl giving rise to mechanical tensions within the tissue of these organs. Host cell wall extensibility and recomposition accompany the growth of the gall and influence pathogen spread and also pathogen life cycle progression. Demethylation of pectin within the extracellular matrix may play an important role in -driven hypertrophy of host underground organs. Through proteomic analysis of the cell wall, we identified proteins accumulating in the galls developing on the underground parts of plants infected with . One of the key proteins identified was the pectin methylesterase (PME18); we further characterized its expression and conducted functional and anatomic studies in the knockout mutant and used Raman spectroscopy to study the status of pectin in -infected galls. We found that late stages of gall formation are accompanied with increased levels of PME18. We have also shown that the massive enlargement of cells colonized with coincides with decreases in pectin methylation. In knockout mutants, could still induce demethylation; however, the galls in this line were smaller and cellular expansion was less pronounced. Alteration in pectin demethylation in the host resulted in changes in pathogen distribution and slowed down disease progression. To conclude, -driven host organ hypertrophy observed during clubroot disease is accompanied by pectin demethylation in the extracellular matrix. The pathogen hijacks endogenous host mechanisms involved in cell wall loosening to create an optimal cellular environment for completion of its life cycle and eventual release of resting spores facilitated by degradation of demethylated pectin polymers.
PubMed: 34394168
DOI: 10.3389/fpls.2021.711838