The Translational Role of MUC8 in Salivary Glands: A Potential Biomarker for Salivary Stone Disease?

Mucin (MUC) 8 has been shown to play an important role in respiratory disease and inflammatory responses. In the present study, we investigated the question of whether MUC8 is also produced and secreted by salivary glands and whether it may also play a role in the oral cavity in the context of inflammatory processes or in the context of salivary stone formation. Tissue samples from parotid and submandibular glands of body donors (n = 6, age range 63–88 years), as well as surgically removed salivary stones from patients (n = 38, age range 48–72 years) with parotid and submandibular stone disease were immunohistochemically analyzed targeting MUC8 and TNFα. The presence of MUC8 in salivary stones was additionally analyzed by dot blot analyses. Moreover, saliva samples from patients (n = 10, age range 51–72 years), who had a salivary stone of the submandibular gland on one side were compared with saliva samples from the other “healthy” side, which did not have a salivary stone, by ELISA. Positive MUC8 was detectable in the inter- and intralobular excretory ducts of both glands (parotid and submandibular). The glandular acini showed no reactivity. TNFα revealed comparable reactivity to MUC8 in the glandular excretory ducts and also did not react in glandular acini. Salivary stones demonstrated a characteristic distribution pattern of MUC8 that differed between parotid and submandibular salivary stones. The mean MUC8 concentration was 71.06 ng/mL in female and 33.21 ng/mL in male subjects (p = 0.156). Saliva from the side with salivary calculi contained significantly (15-fold) higher MUC8 concentration levels than saliva from the healthy side (p = 0.0005). MUC8 concentration in salivary stones varied from 4.59 ng/mL to 202.83 ng/mL. In females, the MUC8 concentration in salivary stones was significantly (2.3-fold) higher, with an average of 82.84 ng/mL compared to 25.27 ng/mL in male patients (p = 0.034). MUC8 is secreted in the excretory duct system of salivary glands and released into saliva. Importantly, MUC8 salivary concentrations vary greatly between individuals. In addition, the MUC8 concentration is gender-dependent (♀️ > ♂). In the context of salivary stone diseases, MUC8 is highly secreted in saliva. The findings support a role for MUC8 in the context of inflammatory events and salivary stone formation. The findings allow conclusions on a gender-dependent component of MUC8.

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Sialotripsy as a treatment option for patients with salivary stone disease

Bulletin of the Medical Institute "REAVIZ" (REHABILITATION, DOCTOR AND HEALTH) ◽

10.20340/vmi-rvz.2021.6.dent.1 ◽

2021 ◽

Author(s):

A. V. Ivachenco ◽

A. E. Yablokov ◽

Е. О. Filatova ◽

V. A. Monakov

Keyword(s):

Salivary Gland ◽

Salivary Glands ◽

Stone Disease ◽

Submandibular Salivary Gland ◽

Clinical Group ◽

Preoperative Stage ◽

Salivary Calculi ◽

Medical University ◽

Methods Of Treatment ◽

Minimally Invasive Methods

Background. The pathologies of the salivary glands are very diverse. Salivary stone disease occupies a special place among diseases of the salivary glands of a non-neoplastic nature. The incidence of salivary stone disease is extremely extensive and accounts for up to 85% of all pathologies of the salivary glands.Objective. Improvement of minimally invasive methods of treatment of salivary stone disease, which does not lead to excision of the gland.Material and methods. On the basis of the Clinics of the Samara State Medical University of the Ministry of Health of Russia, in the period from 2010 to 2020, a clinical group was formed, consisting of 193 patients with salivary stone disease of various localization. The age of the patients ranged from 16 to 84 years. All patients were therapeutically examined at the preoperative stage, voluntary informed consent was signed. Shockwave sialotripsy was performed in 95 patients of the main group (in 64 patients the calculus was located in the submandibular salivary gland, in 31 patients – in the parotid salivary gland). For lithotripsy, we used a lithotripter apparatus "Edap" (France) and "Wolf" (Germany).Results. The analysis of the results obtained as a result of the study made it possible to draw a conclusion about reasonably better results of removal of calculi in the ducts of the salivary glands using the lithotripter "Swiss LithoClast® Master" (Switzerland). At the first visit and the therapeutic effect of the lithotripter, all patients suffering from bouts of salivary colic showed complete cessation.Conclusion. Thus our study, which was devoted to the study of the results of various methods of treating patients with salivary stone disease showed that sialolithotripsy using special narrowly targeted devices for crushing and subsequent excretion of salivary calculi allows to achieve the best results at present.

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Safety profile of the transcription factor EB (TFEB)-based gene therapy through intracranial injection in mice

Translational Neuroscience ◽

10.1515/tnsci-2020-0132 ◽

2020 ◽

Vol 11 (1) ◽

pp. 241-250

Author(s):

Zhenyu Li ◽

Guangqian Ding ◽

Yudi Wang ◽

Zelong Zheng ◽

Jianping Lv

Keyword(s):

Gene Therapy ◽

Transcription Factor ◽

Neurodegenerative Diseases ◽

Brain Tissue ◽

Inflammatory Responses ◽

Tissue Samples ◽

Protein Levels ◽

Virus Serotype ◽

Transcription Factor Eb ◽

The Brain

AbstractTranscription factor EB (TFEB)-based gene therapy is a promising therapeutic strategy in treating neurodegenerative diseases by promoting autophagy/lysosome-mediated degradation and clearance of misfolded proteins that contribute to the pathogenesis of these diseases. However, recent findings have shown that TFEB has proinflammatory properties, raising the safety concerns about its clinical application. To investigate whether TFEB induces significant inflammatory responses in the brain, male C57BL/6 mice were injected with phosphate-buffered saline (PBS), adeno-associated virus serotype 8 (AAV8) vectors overexpressing mouse TFEB (pAAV8-CMV-mTFEB), or AAV8 vectors expressing green fluorescent proteins (GFPs) in the barrel cortex. The brain tissue samples were collected at 2 months after injection. Western blotting and immunofluorescence staining showed that mTFEB protein levels were significantly increased in the brain tissue samples of mice injected with mTFEB-overexpressing vectors compared with those injected with PBS or GFP-overexpressing vectors. pAAV8-CMV-mTFEB injection resulted in significant elevations in the mRNA and protein levels of lysosomal biogenesis indicators in the brain tissue samples. No significant changes were observed in the expressions of GFAP, Iba1, and proinflammation mediators in the pAAV8-CMV-mTFEB-injected brain compared with those in the control groups. Collectively, our results suggest that AAV8 successfully mediates mTFEB overexpression in the mouse brain without inducing apparent local inflammation, supporting the safety of TFEB-based gene therapy in treating neurodegenerative diseases.

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Nutrition and Kidney Stone Disease

Nutrients ◽

10.3390/nu13061917 ◽

2021 ◽

Vol 13 (6) ◽

pp. 1917

Author(s):

Roswitha Siener

Keyword(s):

Kidney Stone ◽

Fluid Intake ◽

Stone Formation ◽

Urinary Stone ◽

Stone Disease ◽

Urinary Stones ◽

Dietary Therapy ◽

Effective Prevention ◽

Kidney Stone Disease ◽

Nutrition And Diet

The prevalence of kidney stone disease is increasing worldwide. The recurrence rate of urinary stones is estimated to be up to 50%. Nephrolithiasis is associated with increased risk of chronic and end stage kidney disease. Diet composition is considered to play a crucial role in urinary stone formation. There is strong evidence that an inadequate fluid intake is the major dietary risk factor for urolithiasis. While the benefit of high fluid intake has been confirmed, the effect of different beverages, such as tap water, mineral water, fruit juices, soft drinks, tea and coffee, are debated. Other nutritional factors, including dietary protein, carbohydrates, oxalate, calcium and sodium chloride can also modulate the urinary risk profile and contribute to the risk of kidney stone formation. The assessment of nutritional risk factors is an essential component in the specific dietary therapy of kidney stone patients. An appropriate dietary intervention can contribute to the effective prevention of recurrent stones and reduce the burden of invasive surgical procedures for the treatment of urinary stone disease. This narrative review has intended to provide a comprehensive and updated overview on the role of nutrition and diet in kidney stone disease.

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Pediatric Urolithiases: A Ten-Year Review

PEDIATRICS ◽

10.1542/peds.65.6.1068 ◽

1980 ◽

Vol 65 (6) ◽

pp. 1068-1072 ◽

Cited By ~ 1

Author(s):

Peter C. Walther ◽

Donald Lamm ◽

George W. Kaplan

Keyword(s):

Recurrence Rate ◽

Stone Formation ◽

Urinary Calculi ◽

Stone Disease ◽

Careful Consideration ◽

Common Type ◽

Presenting Symptoms ◽

Extensive Evaluation ◽

Calculus Disease ◽

High Index Of Suspicion

Sixty-one children who presented with urinary calculi between 1967 and 1977 were reviewed. The overall incidence of calculus disease in our patients was higher than reported previously from other centers in this country. Because presenting symptoms varied, a high index of suspicion was often required for diagnosis. Stones of infection were the most common type identified. Inasmuch as the recurrence rate of calculi in children was low (6.5%), careful consideration of time and cost is warrented before extensive evaluation for metabolic causes of stone formation is undertaken. Immobilization was an important factor in the etiology of stone disease. Most stones less than 5 mm in size passed without surgery.

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Serum Apurinic/Apyrimidinic Endodeoxyribonuclease 1 (APEX1) Level as a Potential Biomarker of Cholangiocarcinoma

Biomolecules ◽

10.3390/biom9090413 ◽

2019 ◽

Vol 9 (9) ◽

pp. 413 ◽

Cited By ~ 3

Author(s):

Doungdean Tummanatsakun ◽

Tanakorn Proungvitaya ◽

Sittiruk Roytrakul ◽

Temduang Limpaiboon ◽

Sopit Wongkham ◽

...

Keyword(s):

Signal Peptide ◽

Transmembrane Helix ◽

Control Group ◽

Cleavage Sites ◽

Dot Blot ◽

Peptide Cleavage ◽

Potential Biomarker ◽

Predicted Signal Peptide ◽

Healthy Control ◽

Dot Blot Analysis

Diagnostic and/or prognostic biomarkers for cholangiocarcinoma (CCA) are still insufficient with poor prognosis of patients. To discover a new CCA biomarker, we constructed our secretome database of three CCA cell lines and one control cholangiocyte cell line using GeLC-MS/MS. We selected candidate proteins by five bioinformatics tools for secretome analysis. The inclusion criteria were as follows: having predicted signal peptide or being predicted as non-classically secreted protein; together with having no transmembrane helix and being previously detected in plasma and having the highest number of signal peptide cleavage sites. Eventually, apurinic/apyrimidinic endodeoxyribonuclease 1 (APEX1) was selected for further analysis. To validate APEX1 as a bio-marker for CCA, serum APEX1 levels of 80, 39, and 40 samples collected from CCA, benign biliary diseases (BBD), and healthy control groups, respectively, were measured using dot blot analysis. The results showed that serum APEX1 level in CCA group was significantly higher than that in BBD or healthy control group. Among CCA patients, serum APEX1 level was significantly higher in patients having metastasis than in those without metastasis. The higher level of serum APEX1 was correlated with the shorter survival time of the patients. Serum APEX1 level might be a diagnostic and prognostic biomarker for CCA.

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CXCL1 contributes to IL-6 expression in osteoarthritis and rheumatoid arthritis synovial fibroblasts by CXCR2, c-Raf, MAPK, and AP-1 pathway

Arthritis Research & Therapy ◽

10.1186/s13075-020-02331-8 ◽

2020 ◽

Vol 22 (1) ◽

Author(s):

Sheng-Mou Hou ◽

Po-Chun Chen ◽

Chieh-Mo Lin ◽

Mei-Ling Fang ◽

Miao-Ching Chi ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Proinflammatory Cytokine ◽

Molecular Mechanisms ◽

Inflammatory Responses ◽

Synovial Fibroblasts ◽

Dependent Manner ◽

Tissue Samples ◽

Chemical Inhibitors ◽

Shrna Plasmid ◽

Signal Cascades

Abstract Background Osteoarthritis (OA) and rheumatoid arthritis (RA) are common joint disorders that are considered to be different diseases due to their unique molecular mechanisms and pathogenesis. Chemokines and their corresponding receptors have been well characterized in RA progression, but less so in OA pathogenesis. Methods The human primary synovial fibroblasts (SFs) were obtained from human OA and RA tissue samples. The Western blot and qPCR were performed to analyze the expression levels of CXCL1, as well as CXCL-promoted IL-6 expression in both OASFs and RASFs. The signal cascades that mediate the CXCL1-promoted IL-6 expression were identified by using chemical inhibitors, siRNAs, and shRNAs. Results Here, we found that both diseases feature elevated levels of CXCL1 and interleukin (IL)-6, an important proinflammatory cytokine that participates in OA and RA pathogenesis. In OASFs and RASFs, CXCL1 promoted IL-6 expression in a dose- and time-dependent manner. In OASFs and RASFs overexpressing CXCL1 or transduced with shRNA plasmid, IL-6 expression was markedly upregulated. CXCR2, c-Raf, and MAPKs were found to regulate CXCL1-induced IL-6 expression in OASFs and RASFs. Finally, CXCL1 triggered the transcriptional activities of c-Jun (which regulates the expression of proinflammatory proteins) in OASFs and RASFs. Conclusions Our present work suggests that the CXCL1/CXCR2 axis helps to orchestrate inflammatory responses in OA and RA SFs.

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Immunolabeling of grapevine flavescence dorée MLO in salivary glands of Euscelidius variegatus: a light and electron microscopy study.

Journal of Histochemistry & Cytochemistry ◽

10.1177/38.1.2294149 ◽

1990 ◽

Vol 38 (1) ◽

pp. 79-85 ◽

Cited By ~ 34

Author(s):

J Lherminier ◽

G Prensier ◽

E Boudon-Padieu ◽

A Caudwell

Keyword(s):

Light Microscopy ◽

Salivary Glands ◽

Light And Electron Microscopy ◽

Microscopy Study ◽

Electron Microscopy Study ◽

Good Tool ◽

Tissue Samples ◽

Thin Sections ◽

Flavescence Dorée ◽

Euscelidius Variegatus

Flavescence dorée (FD), a grapevine yellows disease, is caused by a mycoplasma-like organism (MLO). A colloidal gold indirect immunolabeling technique identified MLO in salivary glands of a vector leafhopper, Euscelidius variegatus. After aldehyde fixation, tissue samples were prepared by cryoultramicrotomy or embedding in acrylic resins. Double fixation with aldehydes and osmium retroxide, followed by embedding in epon, was also performed. Thin or semi-thin serial sections were treated with polyclonal anti-FD-MLO rabbit antibodies, then with gold-conjugated anti-rabbit IgG. Labeling was revealed using the silver enhancement technique for light microscopy. MLO in frozen thin sections of glands were efficiently labeled. Optimal results were obtained with 4% paraformaldehyde-0.1% glutaraldehyde fixation and low-temperature embedding in LR White resin. Both scattered MLO and unusual dense forms of MLO were easily detected with the electron-dense gold probe. This method distinguished MLO from other membrane-limited bodies and provided a good tool for studying infection in large regions of FD-infected tissues by light microscopy.

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Putative Antimicrobial Peptides of the Posterior Salivary Glands from the Cephalopod Octopus vulgaris Revealed by Exploring a Composite Protein Database

Antibiotics ◽

10.3390/antibiotics9110757 ◽

2020 ◽

Vol 9 (11) ◽

pp. 757 ◽

Cited By ~ 1

Author(s):

Daniela Almeida ◽

Dany Domínguez-Pérez ◽

Ana Matos ◽

Guillermin Agüero-Chapin ◽

Hugo Osório ◽

...

Keyword(s):

Antimicrobial Peptides ◽

Salivary Glands ◽

Protein Identification ◽

Inflammatory Responses ◽

Shotgun Proteomics ◽

Octopus Vulgaris ◽

Protein Database ◽

Venom Protein ◽

Protein Toxin ◽

Proteomics Approach

Cephalopods, successful predators, can use a mixture of substances to subdue their prey, becoming interesting sources of bioactive compounds. In addition to neurotoxins and enzymes, the presence of antimicrobial compounds has been reported. Recently, the transcriptome and the whole proteome of the Octopus vulgaris salivary apparatus were released, but the role of some compounds—e.g., histones, antimicrobial peptides (AMPs), and toxins—remains unclear. Herein, we profiled the proteome of the posterior salivary glands (PSGs) of O. vulgaris using two sample preparation protocols combined with a shotgun-proteomics approach. Protein identification was performed against a composite database comprising data from the UniProtKB, all transcriptomes available from the cephalopods’ PSGs, and a comprehensive non-redundant AMPs database. Out of the 10,075 proteins clustered in 1868 protein groups, 90 clusters corresponded to venom protein toxin families. Additionally, we detected putative AMPs clustered with histones previously found as abundant proteins in the saliva of O. vulgaris. Some of these histones, such as H2A and H2B, are involved in systemic inflammatory responses and their antimicrobial effects have been demonstrated. These results not only confirm the production of enzymes and toxins by the O. vulgaris PSGs but also suggest their involvement in the first line of defense against microbes.

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On oral calcifications: sialoliths, dental calculi and tonsilloliths

Microscopy and Microanalysis ◽

10.1017/s1431927613000731 ◽

2013 ◽

Vol 19 (S4) ◽

pp. 23-24 ◽

Cited By ~ 1

Author(s):

A.J. Anjos ◽

P. Nolasco ◽

J.M. Aquino Marques ◽

F. Cabrita ◽

M.F.C. Pereira ◽

...

Keyword(s):

Electron Microscopy ◽

Organic Matter ◽

Calcium Phosphates ◽

Stone Formation ◽

Bacterial Species ◽

Complex Structure ◽

Anaerobic Bacteria ◽

Bacterial Cells ◽

Sulphur Compounds ◽

Salivary Calculi

The oral cavity is susceptible to several calcifications such as salivary calculi (sialoliths), dental calculus (tartar) and tonsillar concretions (tonsilloliths). Although several individual studies had been already carried out, a comprehensive morphological and elemental comparison between them is still missing.Sialoliths are most commonly found in the submandibular glands and are composed of regions rich in Ca and P minerals, namely hydroxyapatite, whitlockite and brushite, and regions consisting of organic matter with high-sulphur content. These regions are organized in alternating concentric layers. Several bacterial species have also been identified in sialoliths microstructure showing that infection occurs recurrently throughout the stone formation.Generally, tartar presents an inorganic structure rich in Ca and P minerals, such as brushite, octacalcium phosphate, hydroxyapatite and whitlockite, and an organic matrix, mainly constituted by aerobic bacteria and yeast or just anaerobic bacteria.Tonsilloliths occur most commonly on the crypts of the palatal tonsils and are composed of a mixture of organic matter, namely bacterial cells and epithelial debris, as well as inorganic material rich in Ca and P minerals such as hydroxyapatite. Volatile sulphur compounds produced by anaerobic bacteria are usually associated to these, in general, innocuous structures.The current study involved the ultrastructure and chemical characterization of the calcified structures by scanning electron microscopy (SEM) combined with energy dispersive spectroscopy carried out with a JEOL JSM 7001F instrument with an INCA pentaFetx3 Oxford spectrometer operated at 15 kV. Higher resolution characterization has been performed by transmission electron microscopy (TEM) using a H8100 Hitachi instrument operated at 200 kV. SEM samples were prepared following metallographic procedures, whereas TEM samples were obtained following standard biological sample preparation procedures.The results show that sialoliths present the most complex structure, with a central core surrounded by concentric layers, while tartar and tonsilloliths do not have a distinctive architecture (Figures 1 (a), 2 (a) and 3 (a). At higher magnifications, layered structures, as well as crystals could be found in sialoliths and tartar (Figures 1 (b) and 2 (b). Bacteria were common in all the calcified structures, although in tonsilloliths their abundance is higher (Figure 3 (b)). All calcifications have similar elemental constitution, with Ca and P, indicating the presence of calcium phosphates (Figures 1 (c), 2 (c) and 3 (c). Sulphur was also found associated with the organic matter in sialoliths and tonsilloliths, though the amounts found in the latter were much smaller than initially expected.Based on the similarities found, new correlations between these calcification will be available. For instance, the mineralization process described in tartar can help understand the similar processes occurring in sialoliths and tonsilloliths, while the association between bacteria and sulphur in tonsilloliths can be a clue for their presence in sialoliths.The work was carried out with financial support of the Portuguese Foundation for Science and Technology through PTDC/SAU-ENB/111941/2009 and PEst-OE/CTM-UI0084/2011 grants.

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Urine protein markers distinguish stone-forming from non-stone-forming relatives of calcium stone formers

AJP Renal Physiology ◽

10.1152/ajprenal.00370.2005 ◽

2006 ◽

Vol 291 (3) ◽

pp. F530-F536 ◽

Cited By ~ 22

Author(s):

Kristin J. Bergsland ◽

Jennifer K. Kelly ◽

Brian J. Coe ◽

Fredric L. Coe

Keyword(s):

Discriminant Analysis ◽

Stone Formation ◽

Stone Disease ◽

Small Subset ◽

Protein Markers ◽

Urine Protein ◽

Multivariate Discriminant Analysis ◽

Stone Formers ◽

Calgranulin B ◽

Multivariate Discriminant

We have investigated urine protein inhibitors of calcium oxalate crystallization to determine whether variations in these proteins are associated with kidney stone disease and whether protein measurements improve the identification of stone formers compared with conventional risk factors (RF). Using Western blotting, we studied variations in the electrophoretic mobility patterns and relative abundances of crystallization-inhibitory proteins in urine from 50 stone-forming (SF) and 50 non-stone-forming (NS) first-degree relatives of calcium SF patients, matched by gender and age. Standard urine chemistry stone risk measurements were also made. Multivariate discriminant analysis was used to test the association of these proteins with nephrolithiasis. Differences in form and abundance of several urine proteins including inter-α-trypsin inhibitor (ITI), prothrombin fragment 1 (PF1), CD59, and calgranulin B (calB) were found to be associated with stone formation. By multivariate discriminant analysis, measurements of forms of PF1, ITI, and calB in men and ITI and CD59 in women, classified 84% of men and 76% of women correctly by stone status. In contrast, standard urine chemistry RF identified only 70% of men correctly and failed to distinguish female SF from NS. Thus a small subset of protein measurements distinguished SF from NS far better than conventional RF in a population of relatives of calcium SF, illustrating the significant association of these proteins with stone disease. Variations in these proteins may serve as markers of stone disease activity or vulnerability to recurrence and may provide new insights into mechanisms of stone formation.

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