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PloS One 2022We imaged the carbohydrate-selective spatial binding of 8 lectins in the ampullary organs (AOs) of electroreceptors on the rostrum of freshwater paddlefish (Polyodon...
We imaged the carbohydrate-selective spatial binding of 8 lectins in the ampullary organs (AOs) of electroreceptors on the rostrum of freshwater paddlefish (Polyodon spathula), by fluorescence imaging and morphometry of frozen sections. A focus was candidate sites of secretion of the glycoprotein gel filling the lumen of AOs. The rostrum of Polyodon is an electrosensory appendage anterior of the head, covered with >50,000 AOs, each homologous with the ampulla of Lorenzini electroreceptors of marine rays and sharks. A large electrosensory neuroepithelium (EN) lines the basal pole of each AO's lumen in Polyodon; support cells occupy most (97%) of an EN's apical area, along with electrosensitive receptor cells. (1) Lectins WGA or SBA labeled the AO gel. High concentrations of the N-acetyl-aminocarbohydrate ligands of these lectins were reported in canal gel of ampullae of Lorenzini, supporting homology of Polyodon AOs. In cross sections of EN, WGA or SBA labeled cytoplasmic vesicles and organelles in support cells, especially apically, apparently secretory. Abundant phalloidin+ microvilli on the apical faces of support cells yielded the brightest label by lectins WGA or SBA. In parallel views of the apical EN surface, WGA labeled only support cells. We concluded that EN support cells massively secrete gel from their apical microvilli (and surface?), containing amino carbohydrate ligands of WGA or SBA, into the AO lumen. (2) Lectins RCA120 or ConA also labeled EN support cells, each differently. RCA120-fluorescein brightly labeled extensive Golgi tubules in the apical halves of EN cells. ConA did not label microvilli, but brightly labeled small vesicles throughout support cells, apparently non-secretory. (3) We demonstrated "sockets" surrounding the basolateral exteriors of EN receptor cells, as candidate glycocalyces. (4) We explored whether additional secretions may arise from non-EN epithelial cells of the interior ampulla wall. (5) Model: Gel is secreted mainly by support cells in the large EN covering each AO's basal pole. Secreted gel is pushed toward the pore, and out. We modeled gel velocity as increasing ~11x, going distally in AOs (toward the narrowed neck and pore), due to geometrical taper of the ampulla wall. Gel renewal and accelerated expulsion may defend against invasion of the AO lumen by microbes or small parasites. (6) We surveyed lectin labeling of accessory structures, including papilla cells in AO necks, striated ectoderm epidermis, and sheaths on afferent axons or on terminal glia.
Topics: Animals; Lectins; Fishes; Bodily Secretions; Microvilli; Epithelial Cells
PubMed: 36395118
DOI: 10.1371/journal.pone.0276854 -
Biochimie Jun 2022Amphibians secrete a complex array of molecules that shape their interactions with coinhabiting microorganisms and macroscopic predators. Glycans are a rapidly evolving...
Amphibians secrete a complex array of molecules that shape their interactions with coinhabiting microorganisms and macroscopic predators. Glycans are a rapidly evolving and complex class of biomolecules implicated in intrinsic and extrinsic recognition events. Despite the numerous studies aiming at the biochemical characterization of anuran skin secretions, little is known about protein-linked oligosaccharides, their synthesis pathways, and their homing secreted glycoproteins. In the present report, LC-MS/MS was used to investigate the diversity of N- and O-linked oligosaccharides in the skin secretion of two South American frogs, Pithecopus azureus and Boana raniceps. Additionally, the enzymes responsible for glycan synthesis pathways were evaluated based on their skin tissue transcriptome. Our analyses allowed the annotation of various N- and O-glycan structures commonly found in vertebrate proteins. Paucimannosidic glycans were abundant in the skin secretion of both amphibians; however, hybrid and complex N-glycan structures were detected only in B. raniceps. A good correlation between the structures discovered in glycomic analyses and transcripts encoding enzymes necessary for their synthesis was obtained. Some transcripts such as those of MAN1A2, FUT8, and ST6GALNAC were found solely in B. raniceps. Finally, secreted N- and O- linked glycoproteins were predicted from the transcriptomic data, indicating that proteases and protease inhibitors are putative sources of the glycans described herein. Overall, our results show the presence of oligosaccharides in amphibians skin secretions and suggest that their diversity is species-specific, paving the way for novel perspectives involving amphibian evolution and ecology.
Topics: Animals; Anura; Chromatography, Liquid; Glycoproteins; Glycosylation; Oligosaccharides; Polysaccharides; Tandem Mass Spectrometry
PubMed: 35077806
DOI: 10.1016/j.biochi.2022.01.008 -
International Journal of Sports... Apr 2021To evaluate the relative importance and predictive ability of salivary immunoglobulin A (s-IgA) measures with regards to upper respiratory illness (URI) in youth...
PURPOSE
To evaluate the relative importance and predictive ability of salivary immunoglobulin A (s-IgA) measures with regards to upper respiratory illness (URI) in youth athletes.
METHODS
Over a 38-week period, 22 youth athletes (age = 16.8 [0.5] y) provided daily symptoms of URI and 15 fortnightly passive drool saliva samples, from which s-IgA concentration and secretion rate were measured. Kernel-smoothed bootstrapping generated a balanced data set with simulated data points. The random forest algorithm was used to evaluate the relative importance (RI) and predictive ability of s-IgA concentration and secretion rate with regards to URI symptoms present on the day of saliva sampling (URIday), within 2 weeks of sampling (URI2wk), and within 4 weeks of sampling (URI4wk).
RESULTS
The percentage deviation from average healthy s-IgA concentration was the most important feature for URIday (median RI 1.74, interquartile range 1.41-2.07). The average healthy s-IgA secretion rate was the most important feature for URI4wk (median RI 0.94, interquartile range 0.79-1.13). No feature was clearly more important than any other when URI symptoms were identified within 2 weeks of sampling. The values for median area under the curve were 0.68, 0.63, and 0.65 for URIday, URI2wk, and URI4wk, respectively.
CONCLUSIONS
The RI values suggest that the percentage deviation from average healthy s-IgA concentration may be used to evaluate the short-term risk of URI, while the average healthy s-IgA secretion rate may be used to evaluate the long-term risk. However, the results show that neither s-IgA concentration nor secretion rate can be used to accurately predict URI onset within a 4-week window in youth athletes.
Topics: Adolescent; Athletes; Humans; Immunoglobulin A; Immunoglobulin A, Secretory; Respiratory Tract Infections; Saliva; Secretory Rate
PubMed: 33440340
DOI: 10.1123/ijspp.2019-0804 -
Current Topics in Developmental Biology 2017The human prostate is a gland of the male reproductive tract, which together with the seminal vesicles, is responsible for most seminal fluid production. It is a common... (Review)
Review
The human prostate is a gland of the male reproductive tract, which together with the seminal vesicles, is responsible for most seminal fluid production. It is a common site of cancer, and unlike other glands, it typically enlarges in aging men. In flies, the male accessory glands make many major seminal fluid components. Like their human equivalents, they secrete proteins from several conserved families, including proteases, lectins, and cysteine-rich secretory proteins, some of which interact with sperm and affect fertility. A key protein, sex peptide, is not conserved in vertebrates but plays a central role in mediating long-term effects on females after mating. Although postmitotic, one epithelial cell type in the accessory glands, the secondary cell, continues to grow in adults. It secretes microvesicles called exosomes from the endosomal multivesicular body, which, after mating, fuse with sperm. They also appear to affect female postmating behavior. Remarkably, the human prostate epithelium also secretes exosomes, which fuse to sperm in vitro to modulate their activity. Exosomes from prostate and other cancer cells are increasingly proposed to play fundamental roles in modulating the tumor microenvironment and in metastasis. Here we review a diverse accessory gland literature, which highlights functional analogies between the male reproductive glands of flies and humans, and a critical role for extracellular vesicles in allowing seminal fluid to promote male interests within the female. We postulate that secondary cells and prostate epithelial cells use common mechanisms to control growth, secretion, and signaling, which are relevant to prostate and other cancers, and can be genetically dissected in the uniquely tractable fly model.
Topics: Animal Structures; Animals; Disease Models, Animal; Drosophila; Humans; Male; Prostate; Prostatic Neoplasms; Semen
PubMed: 28057306
DOI: 10.1016/bs.ctdb.2016.06.001 -
Frontiers in Cellular and Infection... 2019, as a facultative intracellular pathogen, can interact with host macrophages and modulate macrophage function to influence innate and adaptive immunity. Proteins...
, as a facultative intracellular pathogen, can interact with host macrophages and modulate macrophage function to influence innate and adaptive immunity. Proteins secreted by the ESX-1 secretion system are involved in this relationship. Although the importance of ESX-1 in host-pathogen interactions and virulence is well-known, the primary role is ascribed to EsxA (EAST-6) in mycobacterial pathogenesis and the functions of individual components in the interactions between pathogens and macrophages are still unclear. Here, we investigated the effects of EspC on macrophage activation. The EspC protein is encoded by an cluster, which is not linked to the locus, but is essential for the secretion of the major virulence factors of ESX-1, EsxA and EsxB. Our results showed that both EspC protein and EspC overexpression in induced pro-inflammatory cytokines and enhanced surface marker expression. This mechanism was dependent on Toll-like receptor 4 (TLR4), as demonstrated using EspC-treated macrophages from TLR4 mice, leading to decreased pro-inflammatory cytokine secretion and surface marker expression compared with those from wild-type mice. Immunoprecipitation and immunofluorescence assays showed that EspC interacted with TLR4 directly. Moreover, EspC could activate macrophages and promote antigen presentation by inducing mitogen-activated protein kinase (MAPK) phosphorylation and nuclear factor-κB activation. The EspC-induced cytokine expression, surface marker upregulation, and MAPK signaling activation were inhibited when macrophages were blocked with anti-TLR4 antibodies or pretreated with MAPK inhibitors. Furthermore, our results showed that EspC overexpression enhanced the survival of within macrophages and under stress conditions. Taken together, our results indicated that EspC may be another ESX-1 virulence factor that not only modulates the host innate immune response by activating macrophages through TLR4-dependent MAPK signaling but also plays an important role in the survival of pathogenic mycobacteria in host cells.
Topics: Animals; Antigens, Bacterial; Bacterial Proteins; Bodily Secretions; Cytokines; Host-Pathogen Interactions; Humans; Immunity, Innate; Macrophage Activation; Macrophages; Mice; Mice, Inbred C57BL; Mice, Knockout; Mitogen-Activated Protein Kinases; Mycobacterium tuberculosis; Protein C; RAW 264.7 Cells; Signal Transduction; THP-1 Cells; Toll-Like Receptor 2; Toll-Like Receptor 4; Virulence Factors
PubMed: 31134163
DOI: 10.3389/fcimb.2019.00158 -
International Archives of Allergy and... 2020Charcot-Leyden crystal (CLC) protein has been regarded as a hallmark of eosinophilic inflammation.
INTRODUCTION
Charcot-Leyden crystal (CLC) protein has been regarded as a hallmark of eosinophilic inflammation.
OBJECTIVE
The purpose of this study was to investigate the role and levels of CLC protein in patients with nonallergic rhinitis with eosinophilia syndrome (NARES).
METHODS
Overall, 39 NARES patients and 19 controls were recruited. The severity of nasal symptoms was measured by visual analogue scale and serum and local specific immunoglobulin E were determined in all patients. Nasal eosinophilia was assessed by semiquantitative analysis of eosinophils in nasal scrapings. Nasal secretion CLC protein concentrations were evaluated by ELISA.
RESULTS
CLC protein concentrations were significantly higher in NARES patients than in controls (p < 0.0001). Nasal secretion CLC protein levels were significantly correlated with the degree of eosinophilia in nasal scrapings (rs = 0.331; p = 0.04) in NARES patients. Patients with high CLC protein concentrations displayed more severe nasal symptoms than patients with low CLC protein concentrations (p = 0.0080), particularly, nasal itching (p = 0.0029). Pilot study in 8 NARES patients demonstrated that treatment for 1 month with intranasal fluticasone propionate significantly decreased the nasal secretion CLC protein concentrations from baseline levels (p = 0.0335) and markedly attenuated the degree of swelling of inferior turbinate.
CONCLUSIONS
CLC protein levels are significantly higher in nasal secretions of NARES patients and associated with the degree of nasal eosinophilia and the severity of nasal symptoms. Significantly, nasal secretion CLC protein levels obviously decreased after treatment with intranasal corticosteroids, suggesting its possible role in evaluating the medical treatment.
Topics: Adult; Bodily Secretions; Disease Progression; Eosinophilia; Female; Glycoproteins; Humans; Immunoglobulin E; Lysophospholipase; Male; Middle Aged; Nasal Mucosa; Rhinitis; Syndrome; Up-Regulation; Young Adult
PubMed: 32694242
DOI: 10.1159/000509252 -
Journal of Proteomics Aug 2021Metabolome and proteome profiling of biofluids, e.g., urine, plasma, has generated vast and ever-increasing amounts of knowledge over the last few decades.... (Review)
Review
Metabolome and proteome profiling of biofluids, e.g., urine, plasma, has generated vast and ever-increasing amounts of knowledge over the last few decades. Paradoxically, omics analyses of sweat, one of the most readily available human biofluids, have lagged behind. This review capitalizes on the current knowledge and state of the art analytical advances of sweat metabolomics and proteomics. Moreover, current applications of sweat omics such as the discovery of disease biomarkers and monitoring athletic performance are also presented in this review. Another area of emerging knowledge that has been highlighted herein lies in the role of skin host-microbiome interactions in shaping the sweat metabolite-protein profiles. Discussion of future research directions describes the need to have a better grasp of sweat chemicals and to better understand how they function as aided by advances in omics tools. Overall, the role of sweat as an information-rich biofluid that could complement the exploration of the skin metabolome/proteome is emphasized.
Topics: Humans; Metabolome; Metabolomics; Proteome; Proteomics; Sweat
PubMed: 34198014
DOI: 10.1016/j.jprot.2021.104310 -
Scientific Reports Sep 2020Malpighian tubules, analogous to vertebrate nephrons, play a key role in insect osmoregulation and detoxification. Tubules can become infected with a protozoan,...
Malpighian tubules, analogous to vertebrate nephrons, play a key role in insect osmoregulation and detoxification. Tubules can become infected with a protozoan, Malpighamoeba, which damages their epithelial cells, potentially compromising their function. Here we used a modified Ramsay assay to quantify the impact of Malpighamoeba infection on fluid secretion and P-glycoprotein-dependent detoxification by desert locust Malpighian tubules. Infected tubules have a greater surface area and a higher fluid secretion rate than uninfected tubules. Infection also impairs P-glycoprotein-dependent detoxification by reducing the net rhodamine extrusion per surface area. However, due to the increased surface area and fluid secretion rate, infected tubules have similar total net extrusion per tubule to uninfected tubules. Increased fluid secretion rate of infected tubules likely exposes locusts to greater water stress and increased energy costs. Coupled with reduced efficiency of P-glycoprotein detoxification per surface area, Malpighamoeba infection is likely to reduce insect survival in natural environments.
Topics: ATP Binding Cassette Transporter, Subfamily B; ATP Binding Cassette Transporter, Subfamily B, Member 1; Amebiasis; Amoebida; Animals; Biological Transport; Bodily Secretions; Epithelial Cells; Grasshoppers; Infections; Malpighian Tubules; Water-Electrolyte Balance
PubMed: 32994425
DOI: 10.1038/s41598-020-72598-z -
Hepatology (Baltimore, Md.) Sep 2016
Topics: Autophagy; Bile; Cholestasis; Humans; Protein Transport; Secretin
PubMed: 27312229
DOI: 10.1002/hep.28687 -
Microbiome Mar 2023Dental erosion is a disease of the oral cavity where acids cause a loss of tooth enamel and is defined as having no bacterial involvement. The tooth surface is protected...
BACKGROUND
Dental erosion is a disease of the oral cavity where acids cause a loss of tooth enamel and is defined as having no bacterial involvement. The tooth surface is protected from acid attack by salivary proteins that make up the acquired enamel pellicle (AEP). Bacteria have been shown to readily degrade salivary proteins, and some of which are present in the AEP. This study aimed to explore the role of bacteria in dental erosion using a multi-omics approach by comparing saliva collected from participants with dental erosion and healthy controls.
RESULTS
Salivary proteomics was assessed by liquid-chromatography mass spectrometry (LC-MS) and demonstrated two altered AEP proteins in erosion, prolactin inducible protein (PIP), and zinc-alpha-2 glycoprotein (ZAG). Immunoblotting further suggested that degradation of PIP and ZAG is associated with erosion. Salivary microbiome analysis was performed by sequencing the bacterial 16S rRNA gene (V1-V2 region, Illumina) and showed that participants with dental erosion had a significantly (p < 0.05) less diverse microbiome than healthy controls (observed and Shannon diversity). Sequencing of bacterial mRNA for gene expression (Illumina sequencing) demonstrated that genes over-expressed in saliva from erosion participants included H + proton transporter genes, and three protease genes (msrAB, vanY, and ppdC). Salivary metabolomics was assessed using nuclear magnetic resonance spectrometry (NMR). Metabolite concentrations correlated with gene expression, demonstrating that the dental erosion group had strong correlations between metabolites associated with protein degradation and amino acid fermentation.
CONCLUSIONS
We conclude that microbial proteolysis of salivary proteins found in the protective acquired enamel pellicle strongly correlates with dental erosion, and we propose four novel microbial genes implicated in this process. Video Abstract.
Topics: Humans; Tooth Erosion; Proteolysis; Dysbiosis; RNA, Ribosomal, 16S; Saliva; Salivary Proteins and Peptides; Peptide Hydrolases
PubMed: 37004076
DOI: 10.1186/s40168-023-01514-0