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Biomaterials Science Oct 2018The design of new hydrogel-based biomaterials with tunable physical and biological properties is essential for the advancement of applications related to tissue...
The design of new hydrogel-based biomaterials with tunable physical and biological properties is essential for the advancement of applications related to tissue engineering and regenerative medicine. For instance, interpenetrating polymer network (IPN) and semi-IPN hydrogels have been widely explored to engineer functional tissues due to their characteristic microstructural and mechanical properties. Here, we engineered IPN and semi-IPN hydrogels comprised of a tough pectin grafted polycaprolactone (pectin-g-PCL) component to provide mechanical stability, and a highly cytocompatible gelatin methacryloyl (GelMA) component to support cellular growth and proliferation. IPN hydrogels were formed by calcium ion (Ca2+)-crosslinking of pectin-g-PCL chains, followed by photocrosslinking of the GelMA precursor. Conversely, semi-IPN networks were formed by photocrosslinking of the pectin-g-PCL and GelMA mixture, in the absence of Ca2+ crosslinking. IPN and semi-IPN hydrogels synthesized with varying ratios of pectin-g-PCL to GelMA, with and without Ca2+-crosslinking, exhibited a broad range of mechanical properties. For semi-IPN hydrogels, the aggregation of microcrystalline cores led to formation of hydrogels with compressive moduli ranging from 3.1 to 10.4 kPa. For IPN hydrogels, the mechanistic optimization of pectin-g-PCL, GelMA, and Ca2+ concentrations resulted in hydrogels with comparatively higher compressive modulus, in the range of 39 kPa-5029 kPa. Our results also showed that IPN hydrogels were cytocompatible in vitro and could support the growth of three-dimensionally (3D) encapsulated MC3T3-E1 preosteoblasts in vitro. The simplicity, technical feasibility, low cost, tunable mechanical properties, and cytocompatibility of the engineered semi-IPN and IPN hydrogels highlight their potential for different tissue engineering and biomedical applications.
Topics: Biocompatible Materials; Cell Line; Cell Proliferation; Cross-Linking Reagents; Gelatin; Humans; Hydrogels; Materials Testing; Molecular Weight; Pectins; Photochemical Processes; Polyesters; Polymerization; Polymethacrylic Acids; Surface Properties; Tissue Engineering; Tissue Scaffolds
PubMed: 30246835
DOI: 10.1039/c8bm00474a -
Scientific Reports Feb 2018Efficient use of plant-derived materials requires enabling technologies for non-disruptive composition analysis. The ability to identify and spatially locate...
Efficient use of plant-derived materials requires enabling technologies for non-disruptive composition analysis. The ability to identify and spatially locate polysaccharides in native plant tissues is difficult but essential. Here, we develop an optical method for cellulose identification using the structure-responsive, heptameric oligothiophene h-FTAA as molecular fluorophore. Spectrophotometric analysis of h-FTAA interacting with closely related glucans revealed an exceptional specificity for β-linked glucans. This optical, non-disruptive method for stereochemical differentiation of glycosidic linkages was next used for in situ composition analysis in plants. Multi-laser/multi-detector analysis developed herein revealed spatial localization of cellulose and structural cell wall features such as plasmodesmata and perforated sieve plates of the phloem. Simultaneous imaging of intrinsically fluorescent components revealed the spatial relationship between cell walls and other organelles, such as chloroplasts and lignified annular thickenings of the trachea, with precision at the sub-cellular scale. Our non-destructive method for cellulose identification lays the foundation for the emergence of anatomical maps of the chemical constituents in plant tissues. This rapid and versatile method will likely benefit the plant science research fields and may serve the biorefinery industry as reporter for feedstock optimization as well as in-line monitoring of cellulose reactions during standard operations.
Topics: Cell Wall; Cellulose; Glucans; Pectins; Phloem; Plants; Plasmodesmata; Polysaccharides; Stereoisomerism; Thiophenes; Xylans; beta-Glucans
PubMed: 29449697
DOI: 10.1038/s41598-018-21466-y -
Ecotoxicology and Environmental Safety Apr 2022Nitric oxide (NO) is tightly associated with plant response against cadmium (Cd) stress in rice since NO impacts Cd accumulation via modulating cell wall components. In...
Nitric oxide amplifies cadmium binding in root cell wall of a high cadmium-accumulating rice (Oryza sativa L.) line by promoting hemicellulose synthesis and pectin demethylesterification.
Nitric oxide (NO) is tightly associated with plant response against cadmium (Cd) stress in rice since NO impacts Cd accumulation via modulating cell wall components. In the present study, we investigated that whether and how NO regulates Cd accumulation in root in two rice lines with different Cd accumulation ability. The variation of polysaccharides in root cell wall (RCW) of a high Cd-accumulating rice line Lu527-8 and a normal rice line Lu527-4 in response to Cd stress when exogenous NO supplied by sodium nitroprusside (SNP, a NO donor) was studied. Appreciable amounts of Cd distributed in RCW, in which most Cd ions were bound to pectin for the two rice lines when exposed to Cd. Exogenous NO upregulated the expression of OsPME11 and OsPME12 that were involved in pectin demethylesterification, resulting in more low methyl-esterified pectin and therefore stronger pectin-Cd binding. Exogenous NO also enhanced the concentration of hemicellulose and the amount of Cd ions in it. These results demonstrate that NO-induced more Cd binding in RCW in the two rice lines through promoting pectin demethylesterification and increasing hemicellulose accumulation. Higher OsPMEs expression and more hemicellulose synthesis contributed to more Cd immobilization in RCW of the high Cd-accumulating rice line Lu527-8. The main findings of this study reveal the regulation of NO on cell wall polysaccharides modification under Cd stress and help to elucidate the physiological and molecular mechanism of NO participating in Cd responses of rice.
PubMed: 35278988
DOI: 10.1016/j.ecoenv.2022.113404 -
PloS One 2016Tomato (Solanum lycopersicum) is one of the most extensively consumed vegetables but, unfortunately, it is also able to induce allergic reactions. In the past, it has...
Tomato (Solanum lycopersicum) is one of the most extensively consumed vegetables but, unfortunately, it is also able to induce allergic reactions. In the past, it has been shown that the choice of tomato cultivar significantly influenced the allergic reaction of tomato allergic subjects. In this study we investigated the allergenic potential of the cultivated tomato line M82 and of two selected lines carrying small chromosome regions from the wild species Solanum pennellii (i.e. IL7-3 and IL12-4). We evaluated the positive interactions of IgEs of allergic subjects in order to investigate the different allergenic potential of the lines under investigation. We used proteomic analyses in order to identify putative tomato allergens. In addition, bioinformatic and transcriptomic approaches were applied in order to analyse the structure and the expression profiles of the identified allergen-encoding genes. These analyses demonstrated that fruits harvested from the two selected introgression lines harbour a different allergenic potential as those from the cultivated genotype M82. The different allergenicity found within the three lines was mostly due to differences in the IgE recognition of a polygalacturonase enzyme (46 kDa), one of the major tomato allergens, and of a pectin methylesterase (34 kDa); both the proteins were more immunoreactive in IL7-3 compared to IL12-4 and M82. The observed differences in the allergenic potential were mostly due to line-dependent translational control or post-translational modifications of the allergens. We demonstrated, for the first time, that the introgression from a wild species (S. pennellii) in the genomic background of a cultivated tomato line influences the allergenic properties of the fruits. Our findings could support the isolation of favorable wild loci promoting low allergenic potential in tomato.
Topics: Adolescent; Adult; Allergens; Antigens, Plant; Female; Food Hypersensitivity; Gene Expression; Gene Expression Profiling; Genetic Association Studies; Glycosylation; Humans; Immunoglobulin E; Solanum lycopersicum; Male; Proteome; Proteomics; Quantitative Trait Loci; Young Adult
PubMed: 27182705
DOI: 10.1371/journal.pone.0155803 -
Plant Physiology Oct 2018Successful pollen tube elongation is critical for double fertilization, but the biological functions of pollen tube genes and the regulatory machinery underlying this...
Successful pollen tube elongation is critical for double fertilization, but the biological functions of pollen tube genes and the regulatory machinery underlying this crucial process are largely unknown. A previous translatomic study revealed two Arabidopsis () () genes, and , whose expression is up-regulated by pollination. Here, we found that both SAUR62 and SAUR75 localized mainly to pollen tube nuclei. The siliques of homozygous (), (), and the / RNA interference (RNAi) knockdown line had many aborted seeds. These lines had normal pollen viability but defective in vitro and in vivo pollen tube growth, with branching phenotypes. Immunoprecipitation with transgenic SAUR62/75-GFP flowers revealed ribosomal protein RPL12 family members as potential interacting partners, and their individual interactions were confirmed further by yeast two-hybrid and bimolecular fluorescence complementation assays. Polysome profiling showed reduced 80S ribosome abundance in homozygous , , , and RNAi flowers, suggesting that SAUR62/75 play roles in ribosome assembly. To clarify their roles in translation, we analyzed total proteins from RNAi versus wild-type flowers by isobaric tags for relative and absolute quantitation, revealing significantly reduced expression of factors participating in pollen tube wall biogenesis and F-actin dynamics, which are critical for the elastic properties of tube elongation. Indeed, RNAi pollen tubes showed mislocalization of deesterified and esterified pectins and F-actin organization. Thus, the biological roles of SAUR62/75 and their RPL12 partners are critical in ribosomal pre-60S subunit assembly for efficient pollen tube elongation and subsequent fertilization.
Topics: Actins; Amino Acid Sequence; Arabidopsis; Arabidopsis Proteins; Cell Nucleus; Flowers; Gene Expression Regulation, Plant; Genes, Reporter; Intracellular Signaling Peptides and Proteins; Phylogeny; Pollen; Pollen Tube; Pollination; Polyribosomes; Protein Transport; Seeds; Sequence Alignment
PubMed: 30093526
DOI: 10.1104/pp.18.00257 -
Annals of Palliative Medicine Oct 2021Melanoma is derived from malignancies of melanocytes. Anorectal melanoma differs significantly from cutaneous melanoma in clinical presentation, genetic profile, staging...
Melanoma is derived from malignancies of melanocytes. Anorectal melanoma differs significantly from cutaneous melanoma in clinical presentation, genetic profile, staging system, and response to treatment. Anorectal melanoma is seldom diagnosed because most melanoma occurrences are found in the skin tissues. Here, we report 1 case of advanced anorectal melanoma, including its clinical presentation, laboratory findings, imaging, surgical treatment, and pathology. The patient complained of hematochezia and tenesmus. Colonoscopy, computed tomography (CT) scan and digital rectal examination (DRE) revealed a mass near the pectinate line. The patient underwent proctectomy along with colostomy, and subsequent pathological examinations suggested anorectal melanoma with serosa involvement (positive markers: S100, HMB-45, etc.). Evidence-based analyses (single-nucleotide polymorphism (SNPs) and programmed death-ligand 1 (PD-L1) expression) were conducted on the tumor tissue to identify the sensitivity to adjuvant therapies. SNP tests suggested no definite efficacies of commonly used chemotherapeutic agents, with PD-L1 expression implying poor sensitivity to PD-L1 inhibitors. The postoperative recovery was uneventful and the patient was discharged on day 7 after admission. However, the patient refused adjuvant therapies and died 11 months after surgery. In conclusion, anorectal melanoma tends to be mistaken for other common diseases in this region owing to its non-specific clinical presentations. Multidisciplinary treatments are recommended to yield the best possible outcome, despite poor prognosis.
Topics: Anus Neoplasms; B7-H1 Antigen; Chemotherapy, Adjuvant; Drug Resistance, Neoplasm; Fatal Outcome; Humans; Immune Checkpoint Inhibitors; Melanoma; Precision Medicine; Rectal Neoplasms; Skin Neoplasms
PubMed: 34763479
DOI: 10.21037/apm-21-2240 -
Integrative Cancer Therapies Jun 2010To demonstrate the efficacy of PectaSol-C modified citrus pectin (MCP) on prostate cancer in vitro.
AIM
To demonstrate the efficacy of PectaSol-C modified citrus pectin (MCP) on prostate cancer in vitro.
METHOD
Cytotoxicity analysis of PectaSol-C was performed by MTT assay, as were parallel studies with the former brand version of MCP called PectaSol. Apoptosis and inhibition of cell growth were investigated by Western blotting.
RESULTS
Androgen-dependent and -independent human prostate cancer cell lines (LNCaP and PC3, respectively), androgen-dependent and -independent murine prostate cancer cell lines (CASP2.1 and CASP1.1, respectively), as well as noncancerous human benign prostate hyperplasia BPH-1 cell line, were used in the study. MTT assay revealed that 1.0% PectaSol exerted cytotoxicity on LNCaP, PC3, CASP2.1, CASP1.1, and BPH-1 cells for 4-day treatment by 48.0% +/- 2.1%, 54.4% +/- 0.3%, 15.4% +/- 0.8%, 46.1% +/- 1.7%, and 27.4% +/- 1.6%, respectively; whereas 1.0% PectaSol-C showed cytotoxity by 52.2% +/- 1.8%, 48.2% +/- 2.9%, 23.0% +/- 2.6%, 49.0% +/- 1.3%, and 26.8% +/- 2.6%, respectively. Western blotting further confirmed that both MCPs inhibit MAP kinase activation, increase the expression level of its downstream target Bim, a pro-apoptotic protein, and induce the cleavage of Caspase-3 in PC3 and CASP1.1 prostate cancer cells.
CONCLUSION
PectaSol MCP and PectaSol-C MCP can inhibit cell proliferation and apoptosis in prostate cancer cell lines. Our data suggested that 1.0% PectaSol-C can be used for further chemopreventive and chemotherapeutic analysis in vivo.
Topics: Androgens; Animals; Apoptosis; Blotting, Western; Cell Line, Tumor; Cell Proliferation; Citrus; Gene Expression Regulation, Neoplastic; Humans; Male; Mice; Pectins; Prostatic Hyperplasia; Prostatic Neoplasms
PubMed: 20462856
DOI: 10.1177/1534735410369672 -
International Journal of Molecular... Dec 2022Oxidative stress caused by reactive oxygen species (ROS, O2•−, HO•, and H2O2) affects the aging process and the development of several diseases. A new frontier on...
Oxidative stress caused by reactive oxygen species (ROS, O2•−, HO•, and H2O2) affects the aging process and the development of several diseases. A new frontier on its prevention includes functional foods with both specific probiotics and natural extracts as antioxidants. In this work, Panax ginseng C.A. Meyer berries extract was characterized for the presence of beneficial molecules (54.3% pectin-based polysaccharides and 12% ginsenosides), able to specifically support probiotics growth (OD600nm > 5) with a prebiotic index of 0.49. The administration of the extract to a probiotic consortium induced the production of short-chain fatty acids (lactic, butyric, and propionic acids) and other secondary metabolites derived from the biotransformation of Ginseng components. Healthy and tumoral colorectal cell lines (CCD841 and HT-29) were then challenged with these metabolites at concentrations of 0.1, 0.5, and 1 mg/mL. The cell viability of HT-29 decreased in a dose-dependent manner after the exposition to the metabolites, while CCD841 vitality was not affected. Regarding ROS production, the metabolites protected CCD841 cells, while ROS levels were increased in HT-29 cells, potentially correlating with the less functionality of glutathione S-transferase, catalase, and total superoxide dismutase enzymes, and a significant increase in oxidized glutathione.
Topics: Antioxidants; Cell Line, Tumor; Colorectal Neoplasms; Fruit; Hydrogen Peroxide; Panax; Plant Extracts; Prebiotics; Probiotics; Reactive Oxygen Species; Humans; HT29 Cells
PubMed: 36613815
DOI: 10.3390/ijms24010373 -
Case Reports in Surgery 2021A 91-year-old man had a node and erythema in the anal area resistant to treatment. A biopsy of the node in the anus showed atypical cells developing as Paget's disease,...
A 91-year-old man had a node and erythema in the anal area resistant to treatment. A biopsy of the node in the anus showed atypical cells developing as Paget's disease, and staining revealed that the cells were CK7-positive, CK20-positive, and GCDFP15-negative. Therefore, tumor invasion with pagetoid spread (PS) from the anus to the skin was suspected, and the patient was referred to our department for a close examination and surgical treatment. Lower gastrointestinal endoscopy showed edematous, hemorrhagic mucosa in the anal canal, and he was diagnosed with adenocarcinoma via a biopsy. Additionally, redness and swelling with white moss were observed on the skin around the anus. Biopsy showed that Paget cells were diffusely present in the epithelium, and an image of squamous cell carcinoma directly under the epithelium was obtained. Taken together, the patient was diagnosed with the invasion of anal canal cancer with PS to the skin, and we performed laparoscopic abdominoperineal resection and skin carcinoma resection in the perineum. The histopathological analysis showed adenocarcinoma invading the external anal sphincter and subcutaneous adipose tissue in the vicinity of the pectinate line of the anal canal. Pagetoid spread of the adenocarcinoma was observed in the epidermis, and the open portion was slightly invaded up to the rectal mucosa. The anal skin region of the adenocarcinoma partially continued to the hair follicles, and it was complicated by squamous cell carcinoma invading the dermis. There are a few reports of anal canal cancer with PS, and the coexistence of adenocarcinoma and squamous cell carcinoma, as seen in the present case, is rare. We report our case together with relevant literature.
PubMed: 34381623
DOI: 10.1155/2021/9944886 -
Frontiers in Plant Science 2022Pectin, cellulose, and hemicellulose constitute the primary cell wall in eudicots and function in multiple developmental processes in plants. Root hairs are outgrowths...
Pectin, cellulose, and hemicellulose constitute the primary cell wall in eudicots and function in multiple developmental processes in plants. Root hairs are outgrowths of specialized epidermal cells that absorb water and nutrients from the soil. Cell wall architecture influences root hair development, but how cell wall remodeling might enable enhanced root hair formation in response to phosphate (P) deficiency remains relatively unclear. Here, we found that POLYGALACTURONASE INVOLVED IN EXPANSION 2 (PGX2) functions in conditional root hair development. Under low P conditions, a activation tagged line ( ) displays bubble-like root hairs and abnormal callose deposition and superoxide accumulation in roots. We found that the polar localization and trafficking of PIN2 are altered in roots in response to P deficiency. We also found that actin filaments were less compact but more stable in root hair cells and that actin filament skewness in root hairs was recovered by treatment with 1-N-naphthylphthalamic acid (NPA), an auxin transport inhibitor. These results demonstrate that activation tagging of affects cell wall remodeling, auxin signaling, and actin microfilament orientation, which may cooperatively regulate root hair development in response to P starvation.
PubMed: 35586221
DOI: 10.3389/fpls.2022.862171