Saliva Secretion Research Articles

Differences in salivary secretion and epidermal growth factor concentrations in mild reflux esophagitis and non-erosive reflux disease.

Sex differences in salivary secretion have been reported among healthy subjects. In the present study, salivary secretion and salivary epidermal growth factor (EGF) concentrations were investigated in mild reflux esophagitis patients, non-erosive reflux disease (NERD) patients, and healthy controls by matching the sex ratio. Thirty-three (male:female = 11:22) patients with NERD, 33 (11:22) with mild reflux esophagitis, and 33 (11:22) healthy controls were recruited for this case-control study. Salivary secretion was assessed as follows: each patient chewed sugar-free gum for 3min prior to endoscopy, and the amount of saliva secretion, salivary pH, and salivary pH after acid loading as an index of the acid-buffering capacity were measured. Salivary EGF concentrations were measured by ELISA. No significant differences were observed in the amount of saliva secretion, salivary pH, or the acid-buffering capacity between the mild reflux esophagitis and NERD groups. However, the amount of saliva secretion and the acid-buffering capacity in the mild reflux esophagitis group and the amount of saliva secretion, salivary pH, and the acid-buffering capacity in the NERD group were significantly lower than those in the healthy control group. No significant differences were noted in salivary EGF concentrations between the mild reflux esophagitis and NERD groups. After matching the sex ratio, the saliva secretion was significantly lower in patients with mild reflux esophagitis and NERD than in healthy controls. However, no significant differences were observed in the amount of saliva secretion or salivary EGF concentrations between both groups.

Short-term and bystander effects of radiation on murine submandibular glands.

Many patients treated for head and neck cancers experience salivary gland hypofunction due to radiation damage. Understanding the mechanisms of cellular damage induced by radiation treatment is important in order to design methods of radioprotection. In addition, it is crucial to recognize the indirect effects of irradiation and the systemic responses that may alter saliva secretion. In this study, radiation was delivered to murine submandibular glands (SMGs) bilaterally, using a 137Cs gamma ray irradiator, or unilaterally, using a small-animal radiation research platform (SARRP). Analysis at 3, 24 and 48 h showed dynamic changes in mRNA and protein expression in SMGs irradiated bilaterally. Unilateral irradiation using the SARRP caused similar changes in the irradiated SMGs, as well as significant off-target, bystander effects in the non-irradiated contralateral SMGs.

Disturbed natural killer cell homeostasis in the salivary gland enhances autoimmune pathology via IFN-γ in a mouse model of primary Sjögren’s syndrome

Innate lymphoid cells (ILCs), including natural killer (NK) cells, ILC1, ILC2, lymphoid tissue-inducer (LTi) cells, and ILC3 cell, play a key role in various immune responses. Primary Sjögren's syndrome (pSS) is an autoimmune disease characterized by chronic inflammation of exocrine glands, such as the lacrimal and salivary glands (SGs). The role of NK cells among ILCs in the pathogenesis of pSS is still unclear. In this study, the characteristics and subsets of NK cells in the salivary gland (SG) tissue were analyzed using a murine model of pSS. Multiple phenotypes and cytotoxic signature of the SG NK cells in control and pSS model mice were evaluated by flow cytometric analysis. Intracellular expression of interferon-γ (IFN-γ) among T cells and NK cells from the SG tissues was compared by in vitro experiments. In addition, pathological analysis was performed using anti-asialo-GM1 (ASGM1) antibody (Ab)-injected pSS model mice. The number of conventional NK (cNK) cells in the SG of pSS model mice significantly increased compared with that in control mice at 6 weeks of age. The production level of IFN-γ was significantly higher in SG NK cells than in SG T cells. The depletion of NK cells by ASGM1 Ab altered the ratio of tissue resident NK (rNK) cells to cNK cells, which inhibited the injury to SG cells with the recovery of saliva secretion in pSS model mice. The results indicate that SG cNK cells may enhance the autoreactive response in the target organ by upregulating of IFN-γ, whereas SG rNK cells protect target cells against T cell cytotoxicity. Therefore, the activation process and multiple functions of NK cells in the target organ could be helpful to develop potential markers for determining autoimmune disease activity and target molecules for incurable immune disorders.

Simulation of Calcium Dynamics in Realistic Three-Dimensional Domains.

The cytosolic concentration of free calcium ions () is an important intracellular messenger in most cell types, and the spatial distribution of is often critical. In a salivary gland acinar cell, a polarised epithelial cell, whose principal function is to transport water and thus secrete saliva, controls the secretion of primary saliva, but increases in are localised to the apical regions of the cell. Hence, any quantitative explanation of how controls saliva secretion must take into careful account the spatial distribution of the various sources, sinks, and -sensitive ion channels. Based on optical slices, we have previously constructed anatomically accurate three-dimensional models of seven salivary gland acinar cells, and thus shown that a model in which responses are confined to the apical regions of the cell is sufficient to provide a quantitative and predictive explanation of primary saliva secretion. However, reconstruction of such anatomically accurate cells is extremely time consuming and inefficient. Here, we present an alternative, mostly automated method of constructing three-dimensional cells that are approximately anatomically accurate and show that the new construction preserves the quantitative accuracy of the model.

A recombinase polymerase amplification (RPA) combined with strip visualization method for RNA-based presumptive tests of saliva and vaginal secretion

Identifying the origin of body fluids is a critical step in a forensic investigation. One widely used method to identify human body fluids is based on the color visualization of immune antigen detection strips for detecting hemoglobin in blood and prostate-specific antigen in semen. It is highly imperative to construct an easy-to-perform, mRNA-based method for the point-of-care identification of other human body fluids, such as saliva and vaginal secretion. Here, we established specific strips with the mRNA markers STATH (for saliva) and SPINK5 (for vaginal secretion) via reverse transcription recombinase polymerase amplification (RT-RPA) and lateral flow dipstick (LFD) assays (RT-RPA-LFD). RT-RPA could be accomplished in a single tube at a wide temperature range of 30–42 ℃ within 10–25 min if we do not count time for RNA extraction. The diluted RPA products were added onto the LFD strip pad to visually observe the color change of the Control/Test line. The tissue specificity and detection limit of the assays were evaluated using the optimized reaction conditions of RPA at 37 ℃ for 15 min. The positive signals of STATH were observed both in saliva and nasal secretions. SPINK5 was positive in a template-dependent manner in 4 out of 30 female urine samples in addition to vaginal secretion and menstrual blood samples. Cross-reactions were not detected in semen, skin swabs, sweat, or male urine. Both assays were capable of detecting aged samples, which were stored for 180 days (saliva) or 300 days (vaginal secretion) at room temperature. Moreover, saliva or vaginal secretion was successfully detected in all kinds of mixtures made from various body fluids. Overall, the rapid strip test method by the RT-RPA-LFD assay is simple, time-saving and highly sensitive for estimating the tissue origin of saliva and vaginal secretion. This method for the rapid RNA-based presumptive tests of the tissue type of body fluids is easy to perform prior to a multiplex mRNA analysis, which can demonstrate more reliable saliva or vaginal secretion identification.

DMA, a Small Molecule, Increases Median Survival and Reduces Radiation-Induced Xerostomia via the Activation of the ERK1/2 Pathway in Oral Squamous Cell Carcinoma.

Simple SummaryRadiotherapy is commonly used to treat the majority of patients with head and neck cancers. Salivary glands in the radiation field are affected by this procedure. The purpose of this study was to investigate the role of DMA as a radiomodulator to evaluate the real possibilities of reducing the incidence and severity of xerostomia in head and neck squamous cell carcinoma (HNSCC) patients. The effect of DMA on the response of head and neck cell lines in the presence of radiation was analyzed using clonogenic survival and xenograft tumor assays for cell line-derived xenografts as well as patient-derived xenografts. The combinatorial treatment of DMA along with radiation influenced the migration of cells. The changes associated with migration were observed through a microscope and wound healing assay. In addition, the stimulated saliva was measured. Tissue sections were analyzed using immunohistochemistry to stain for molecular markers of cell proliferation and porin channels.Survival, recurrence, and xerostomia are considerable problems in the treatment of oral squamous carcinoma patients. In this study, we investigated the role of DMA (5-(4-methylpiperazin-1-yl)-2-[2′-(3,4-dimethoxyphenyl)5″benzimidazoyl]benzimidazole) as a salivary gland cytoprotectant in a patient-derived xenograft mouse model. A significant increase in saliva secretion was observed in the DMA-treated xenograft compared to radiation alone. Repeated doses of DMA with a high dose of radiation showed a synergistic effect on mice survival and reduced tumor growth. The mean survival rate of tumor-bearing mice was significantly enhanced. The increased number of Ki-67-stained cells in the spleen, intestine, and lungs compared to the tumor suggests DMA ablates the tumor but protects other organs. The expression of aquaporin-5 was restored in tumor-bearing mice injected with DMA before irradiation. The reduced expression of αvβ3 integrin and CD44 in DMA alone and DMA with radiation-treated mice suggests a reduced migration of cells and stemness of cancer cells. DMA along with radiation treatment results in the activation of the Ras/Raf/MEK/ERK pathway in the tumor, leading to apoptosis through caspase upregulation. In conclusion, DMA has strong potential for use as an adjuvant in radiotherapy in OSCC patients.

STATE OF ORAL HYGIENE, RATE OF SALIVATION AND VISCOSITY OF ORAL LIQUID IN 3-6 YEARS OLD AGED CHILDREN, TAKING INTO ACCOUNT THE DEGREE OF CARIES ACTIVITY

Oral fluid performs a number of important functions: mineralizing, protective, cleansing, creating an optimal environment for the functioning of the hard tissues of the teeth. Research results show that the condition of the oral cavity, its biophysical and biochemical indicators provide information on the general resistance of the body. The possibility of complicating both dental diseases and somatic pathology at the expense of each other has been proven, that is, there is a mutual aggravation. The indicators were studied taking into account the age of the children and the degree of caries activity in them. 335 children aged 3-6 years were examined. Oral hygiene index according to Fedorov-Volodkina was determined in each age period and taking into account the degree of caries activity. Thus, children without caries and children with a compensated degree of caries activity in all age groups had a satisfactory state of oral hygiene. A comparison of 3-year-old children, but of different groups by the degree of caries activity, did not reveal a probable difference in the state of hygiene. And in 4, 5, and 6-year-olds with subcompensated and decompensated levels of caries activity, a probable difference in the state of oral hygiene was revealed. We observe a significant increase in the hygiene index in the entire group of children aged 3-6 years, respectively, with an increase in the Decay Extracted Filling index (р < 0,01; р < 0,001). The rate of salivation in the examined children of all groups corresponded to the norm or was at the level of slight hyposalivation. During the examination, a significant decrease in the numerical values of the salivary rate was found in children with subcompensated and decompensated caries activity compared to healthy children and with compensated caries activity. Studying the viscosity index of oral fluid, a probable increase in the index was established in children with subcompensated and decompensated levels of caries activity compared to healthy children in each age group. In children with a decompensated degree of caries activity, the highest index of oral fluid viscosity was found in each age group.
 The results of the examination showed a significant deterioration of oral hygiene in children with various degrees of caries activity compared to children without caries. The rate of saliva secretion is also likely to decrease, and the viscosity of the oral fluid increases as the caries situation in the oral cavity worsens in all age groups. Considering the multifactorial etiology of caries, it is necessary to continue the study of the relationship between the occurrence of caries in children, taking into account the influence of oral fluid and morphological features of the occlusal surface on the state of resistance of the hard tissues of the teeth.

Mitochondrial double-stranded RNAs as a pivotal mediator in the pathogenesis of Sjӧgren’s syndrome

Sjӧgren’s syndrome (SS) is a systemic autoimmune disease that targets the exocrine glands, resulting in impaired saliva and tear secretion. To date, type I interferons (I-IFNs) are increasingly recognized as pivotal mediators in SS, but their endogenous drivers have not been elucidated. Here, we investigate the role of mitochondrial double-stranded RNAs (mt-dsRNAs) in regulating I-IFNs and other glandular phenotypes of SS. We find that mt-dsRNAs are elevated in the saliva and tears of SS patients (n = 73 for saliva and n = 16 for tears) and in salivary glands of non-obese diabetic mice with salivary dysfunction. Using the in-house-developed 3D culture of immortalized human salivary gland cells, we show that stimulation by exogenous dsRNAs increase mt-dsRNAs, activate the innate immune system, trigger I-IFNs, and promote glandular phenotypes. These responses are mediated via the Janus kinase 1 (JAK1)/signal transducer and activator of transcription (STAT) pathway. Indeed, a small chemical inhibitor of JAK1 attenuates mtRNA elevation and immune activation. We further show that muscarinic receptor ligand acetylcholine ameliorates autoimmune characteristics by preventing mt-dsRNA-mediated immune activation. Last, direct suppression of mt-dsRNAs reverses the glandular phenotypes of SS. Altogether, our study underscores the significance of mt-dsRNA upregulation in the pathogenesis of SS and suggests mt-dsRNAs as propagators of a pseudo-viral signal in the SS target tissue.

Aerosolized Zika virus infection in Guinea pigs

ABSTRACT Zika virus (ZIKV) is primarily transmitted through mosquito bites and sexual contact, and vertical transmission of ZIKV has also been observed in humans. In addition, ZIKV infection via unknown transmission routes has been frequently reported in clinical settings. However, whether ZIKV can be transmitted via aerosol routes remains unknown. In this study, we demonstrated that aerosolized ZIKV is fully infectious in vitro and in vivo. Remarkably, intratracheal (i.t.) inoculation with aerosolized ZIKV led to rapid viremia and viral secretion in saliva, as well as robust humoral and innate immune responses in guinea pigs. Transcriptome analysis further revealed that the expression of genes related to viral processes, biological regulation and the immune response was significantly changed. Together, our results confirm that aerosolized ZIKV can result in systemic infection and induce both innate and adaptive immune responses in guinea pigs, highlighting the possibility of ZIKV transmission via aerosols.

Nicotinamide mitigates radiation injury in submandibular gland by protecting mitochondrial structure and functions.

Radiation damage to salivary gland is inevitable in head and neck cancer patients receiving radiotherapy. Safe and effective treatments for protecting salivary glands from radiation are still unavailable. Mitochondrial damage is a critical mechanism in irradiated salivary gland; however, treatment targeting mitochondria has not received much attention. Nicotinamide is a key component of the mitochondrial metabolism. Here, we investigated the effects and underlying mechanisms of nicotinamide on protecting irradiated submandibular gland. Submandibular gland cells and tissues were randomly divided into four groups: control, nicotinamide alone, radiation alone, and radiation with nicotinamide pretreatment. Cell viability was detected by PrestoBlue cell viability reagent. Histopathological alterations were observed with HE staining. Pilocarpine-stimulated saliva was measured from Wharton's duct. Cell apoptosis was determined by flow cytometry and terminal deoxynucleotidyl transferase dUTP nick-end labeling assay. Nicotinamide phosphoribosyl transferase was examined with immunofluorescence. The levels of nicotinamide adenine dinucleotide, mitochondrial membrane potential, and adenosine triphosphate were measured with the relevant kits. The mitochondrial ultrastructure was observed under transmission electron microscopy. Nicotinamide significantly mitigated radiation damage both in vitro and in vivo. Also, nicotinamide improved saliva secretion and reduced radiation-induced apoptosis in irradiated submandibular glands. Moreover, nicotinamide improved nicotinamide phosphoribosyl transferase and the levels of nicotinamide adenine dinucleotide/adenosine triphosphate and mitochondrial membrane potential, all of which were decreased by radiation in submandibular gland cells. Importantly, nicotinamide protected the mitochondrial ultrastructure from radiation. These findings demonstrate that nicotinamide alleviates radiation damage in submandibular gland by replenishing nicotinamide adenine dinucleotide and maintaining mitochondrial function and ultrastructure, suggesting that nicotinamide could be used as a prospective radioprotectant for preventing radiation sialadenitis.

A Comprehensive Characterization of Small RNA Profiles by Massively Parallel Sequencing in Six Forensic Body Fluids/Tissue.

Body fluids/tissue identification (BFID) is an essential procedure in forensic practice, and RNA profiling has become one of the most important methods. Small non-coding RNAs, being expressed in high copy numbers and resistant to degradation, have great potential in BFID but have not been comprehensively characterized in common forensic stains. In this study, the miRNA, piRNA, snoRNA, and snRNA were sequenced in 30 forensic relevant samples (menstrual blood, saliva, semen, skin, venous blood, and vaginal secretion) using the BGI platform. Based on small RNA profiles, relative specific markers (RSM) and absolute specific markers (ASM) were defined, which can be used to identify a specific body fluid/tissue out of two or six, respectively. A total of 5204 small RNAs were discovered including 1394 miRNAs (including 236 novel miRNA), 3157 piRNAs, 636 snoRNAs, and 17 snRNAs. RSMs for 15 pairwise body fluid/tissue groups were discovered by differential RNA analysis. In addition, 90 ASMs that were specifically expressed in a certain type of body fluid/tissue were screened, among them, snoRNAs were reported first in forensic genetics. In brief, our study deepened the understanding of small RNA profiles in forensic stains and offered potential BFID markers that can be applied in different forensic scenarios.

Ascorbic acid induces salivary gland function through TET2/acetylcholine receptor signaling in aging SAMP1/Klotho (-/-) mice.

Aging affects salivary gland function and alters saliva production and excretion. This study aimed to investigate whether ascorbic acid can be used to treat salivary gland dysfunction in an extensive aging mouse model of SAMP1/Klotho-/- mice. In our previous study, we found that ascorbic acid biosynthesis was disrupted in the salivary glands of SAMP1/Klotho (-/-) mice subjected to metabolomic profiling analysis. In SAMP1/Klotho -/- mice, daily supplementation with ascorbic acid (100 mg/kg for 18 days) significantly increased saliva secretion compared with the control. The expression of salivary gland functional markers (α-amylase, ZO-1, and Aqua5) is upregulated. Additionally, acetylcholine and/or beta-adrenergic receptors (M1AchR, M3AchR, and Adrb1) were increased by ascorbic acid in the salivary glands of aging mice, and treatment with ascorbic acid upregulated the expression of acetylcholine receptors through the DNA demethylation protein TET2. These results suggest that ascorbic acid could overcome the lack caused by dysfunction of ascorbic acid biosynthesis and induce the recovery of salivary gland function.

<em>In Vivo</em> Vascular Permeability Detection in Mouse Submandibular Gland

Saliva plays an important role in oral and overall health. The intact endothelial barrier function of blood vessels enables saliva secretion, whereas the endothelial barrier dysfunction is related to many salivary gland secretory disorders. The present protocol describes an in vivo paracellular permeability detection method to evaluate the function of endothelial tight junctions (TJs) in mouse submandibular glands (SMG). First, fluorescence-labeled dextrans with different molecular weights (4 kDa, 40 kDa, or 70 kDa) were injected into the angular veins of mice. Afterward, the unilateral SMG was dissected and fixed in the customized holder under a two-photon laser-scanning microscope, and then images were captured for blood vessels, acini, and ducts. Utilizing this method, the real-time dynamic leakage of the different-sized tracers from blood vessels into the basal sides of the acini and even across the acinar epithelia into the ducts was monitored to evaluate the alteration of the endothelial barrier function under physiological or pathophysiological conditions.

The Influence of Periodontal Diseases and the Stimulation of Saliva Secretion on the Course of the Acute Phase of Ischemic Stroke.

Background and purpose: The course of an ischemic stroke depends on many factors. The influence of periodontal diseases and the stimulation of salivation on the course and severity of stroke remains unresolved. Therefore, the aim of the study was to analyze the severity of ischemic stroke depending on the occurrence of periodontal diseases and saliva stimulation. Methods: The severity of the neurological condition was assessed using the NIHSS scale on days one, three and seven of stroke. The incidence of periodontal diseases was classified using the Hall’s scale in the first day of stroke. On days one and seven of stroke, the concentration of IL-1β, MMP-8, OPG and RANKL in the patients’ saliva was assessed using the Elisa technique. At the same time, the level of CRP and the number of leukocytes in the peripheral blood were tested on days one, three and seven of the stroke, and the incidence of upper respiratory and urinary tract infections was assessed. Results:100 consecutive patients with their first ever ischemic stroke were enrolled in the study. 56 randomly selected patients were subjected to the stimulation of salivation, the remaining patients were not stimulated. In the study of the severity of the neurological condition using the NIHS scale on days three and seven of stroke, the degree of deficit in patients without periodontal disease significantly improved compared to patients with periodontal disease, respectively (p < 0.01 and p = 0.01). Patients from the stimulated group had more severe neurological deficit at baseline (p = 0.04). On days three and seven of neurological follow-up, the condition of patients from both groups improved with a further distinct advantage of the unstimulated group over the stimulated group, respectively (p = 0.03 and p < 0.001). In patients from both groups, a statistically significant decrease in CRP and lymphocyte levels was observed on day seven in relation to day one. Conclusions: The occurrence of periodontal disease in a patient with stroke affects the severity of stroke. Stimulation of the mouth and salivary glands in these patients may have a positive effect on the course of stroke, taking into account the dynamics of neurological symptoms.

Oral changes in patients with psoriasis.

Psoriasis is one of the most frequent skin diseases. The cause of psoriasis is not fully expained as there are many factors (infectious, traumatic, hormonal, and chemical) that may play a role in the manifestation of its symptoms. One of the factors that may contribute to the appearance of psoriatic lesions may be the lesions in the oral cavity. The occurrence of lesions in the oral cavity is defined as rare, what can be explained by their nonspecific clinical image, and also by the possibility of being overlooked. Most characteristic symptoms of psoriasis occurring in the oral cavity are the geographic tongue and fissured tongue. It is also believed that there is a correlation between psoriasis and oral health- the periodontal and teeth condition as well as changes in saliva secretion. The psoriasis arthritis changes can also affect temporomandibular joint and impair the function of stomatognathic system. Because of these reports, cooperation of dermatologists and dentists in psoriasis care seems to be necessary.

A Mathematical Model of Salivary Gland Duct Cells

Saliva is produced in two stages in the salivary glands: the secretion of primary saliva by the acinus and the modification of saliva composition to final saliva by the intercalated and striated ducts. In order to understand the saliva modification process, we develop a mathematical model for the salivary gland duct. The model utilises the realistic 3D structure of the duct reconstructed from an image stack of gland tissue. Immunostaining results show that TMEM16A and aquaporin are expressed in the intercalated duct cells and that ENaC is not. Based on this, the model predicts that the intercalated duct does not absorb Na^+ and Cl^- like the striated duct but secretes a small amount of water instead. The input to the duct model is the time-dependent primary saliva generated by an acinar cell model. Our duct model produces final saliva output that agrees with the experimental measurements at various stimulation levels. It also shows realistic biological features such as duct cell volume, cellular concentrations and membrane potentials. Simplification of the model by omission of all detailed 3D structures of the duct makes a negligible difference to the final saliva output. This shows that saliva production is not sensitive to structural variation of the duct.

Immunohistochemical Evaluation of the Pathological Effects of Diabetes Mellitus on the Major Salivary Glands of Albino Rats.

Diabetes mellitus is a notorious chronic disease characterized by hyperglycemia. Our study aimed to determine the expression of cytokeratin 17 (CK17) in all major salivary glands of diabetic albino rats to provide more information about the pathological effects of DM on the intracellular structures of the gland parenchyma. Twenty male adult albino rats were utilized in the experiment and divided into two equal groups, group 1 (control rats) and group 2 (diabetic rats). The animals were sacrificed 45 days after diabetes induction. The major salivary gland complex of all groups was dissected and prepared for evaluation by histological and immunohistochemical expression of CK17. Histological results prove that the salivary gland parenchyma of diabetic group undergo gland atrophy characterized with the presence of degenerated acini, dilated duct system, and presence of duct-like structure with predominance of fibrous tissue compartment and discrete fat cells. Immunohistochemical expression of CK17 of major salivary gland of control group revealed negative to diffuse mild expression in all duct cells and some serous acinar cells, whereas mucous acini were negatively stained. On the other hand, major salivary gland parenchyma of diabetic group demonstrated mild to strong expression of duct cells more concentrated at their apical part with moderate to strong expression of some serous acini of diffuse type, whereas mucous acini of both submandibular gland and sublingual gland were negatively stained. The severity and prevalence of CK 17 in our results are predictive of the pathological influence of the DM that interferes with saliva production and/or secretion leading to dry mouth. The results also showed clear changes in the cytokeratin expression of diabetic sublingual salivary gland, although it had little effect in the routine histological study with hematoxylin and eosin, confirming that routine studies are not sufficient to form a definitive opinion.

Comparative Study of Cytokeratin Immunostaining of Parotid Gland Parenchyma in Normal, Diabetic, and Excretory Duct Ligation of Mongrel Dogs.

The present study aimed to give a glimpse of the normal distribution of intermediate filaments within the parotid gland parenchyma of mongrel dogs and to reveal the pathological changes that may occur as a result of the effects of diabetes mellitus or atrophy of the gland caused by the ligation of the excretory duct to discover whether there is a similarity in these pathological behaviors. Twelve healthy mongrel dogs were used in the experiment and were divided into three groups: group I (the control group), group II (dogs with alloxan-induced diabetes), and group III (dogs with the right-side duct-ligated parotid gland). The dogs were sacrificed 45 days after the parotid excretory duct were tied. The right parotid gland of all groups was dissected and prepared for histological and immunohistochemical expression of cytokeratin 17 assay. Histological findings confirmed that the parotid gland parenchyma of the diabetic group had glandular atrophy characterized by the loss of gland structure, degenerated acini, and dilatation of the duct system. Moreover, there is a predominance of the fibrous component with the presence of fat cells within the gland compartments. On the contrary, the excretory duct-ligated group undergoes severe glandular atrophy of the previous character with the presence of duct-like structure as well as extravasation and vasodilatation. Immunohistochemical expression of cytokeratin 17 in control parotid using an immunoperoxidase technique showed that cytokeratin expression varies from negative to mild in all ducts and some serous acinar cells. The gland parenchyma of the diabetic group showed mild to strong cytokeratin expression of duct cells more concentrated in the apical part with moderate to strong expression of diffuse type in some serous acini. The intensity of cytokeratin 17 in gland compartments of the excretory duct-ligated group revealed a variation in expression that ranged from negative to strong diffuse staining throughout the gland. The severity and prevalence of cytokeratin 17 in our results are predictive of the pathological influence of both diabetes mellitus and duct ligation on the cytokeratin intracellular filaments of the salivary gland parenchyma in a different way that interferes with saliva production and/or secretion leading to xerostomia.

Abstract 2797: Proteomic signature in exhaled breath condensates for a non-invasive diagnostics of lung cancer

Abstract Because of the invasiveness of examination methods as bronchoscopy for the diagnosis of lung cancer, there is currently no preventive screening for this disease. Thus, lung cancer is usually diagnosed in stage 3 or 4, when it is too late for successful treatment. However, if diagnosed early, it can be cured entirely. Therefore, it is necessary to find a suitable method to screen the high-risk populations and reveal lung cancer in the early stages. Since the mucous membranes of the respiratory tract are naturally moistened by fluids containing proteins, metabolites, chemoattractants and growth factors released by the highly active epithelium, the collection of the respiratory tract fluids can be used to diagnose malignant and benign respiratory tract diseases. The most commonly used method in this sense is bronchoalveolar lavage. However, the technique itself is not pleasant and can be accompanied by possible complications. Therefore, intensive research work is conducted to develop non-invasive methods for biomarker detection in exhaled breath, sputum, saliva or nasal secretion. We have studied an exhaled breath condensate (EBC) matrix, which is collected non-invasively and the patients are not suffering from its collection. Several studies have reported proteomic analysis of EBC while identifying low hundreds of proteins by mass spectrometry (MS). We have developed a potent gel-free MS-based approach for sample preparation and analysis. Together with a powerful search tool, we were able to identify a ten times higher number of proteins across a comparable group of individuals of about three hundred peoples’ samples. Our approach seems to be highly reproducible and applicable to various respiratory and systemic diseases. In our work focused on non-small lung cancer biomarkers compared to chronic obstructive pulmonary disease (COPD) and healthy controls, we have identified 7694 proteins and of them, 6525 proteins were quantified at least in one replicate across 296 individuals’ samples measured in triplicates. Combining univariate and multivariate statistical approaches and sensitivity analysis, we have suggested biomarkers that could distinguish lung cancer patients from COPD and healthy individuals. This research was supported by grants from the Czech Ministry of Education, Youth and Sports (EATRIS-CZ - LM2018133), European Regional Development Fund - Project ENOCH (No. CZ.02.1.01/0.0/0.0/16_019/0000868) and IGA_LF_2021_036 (Palacky University in Olomouc). Citation Format: Jana Vaclavkova, Jana Vrbkova, Pavla Kourilova, Dusan Holub, Juraj Kultan, Petr Jakubec, Ondrej Fischer, Frantisek Kopriva, Vendula Latalova, Tatiana Gvozdiakova, Marian Hajduch, Petr Dzubak. Proteomic signature in exhaled breath condensates for a non-invasive diagnostics of lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2797.

Review on Spilanthe sacmella

Spilanthes acmella is a medicinal plant that is mostly found in sub-tropical and tropical regions worldwide. It is also known as toothache plant as it is known to reduce pain related to toothaches. Various studies have reported that it possesses numerous biological properties including anti-inflammatory, antimicrobial, antipyretic, local anesthetic, antioxidantand analgesic effects. It has traditional uses in health care and in cuisines as a flavor. Its flowers and leaves can be eaten and causes saliva secretion and tingling sensation in the tongue. Spilanthol is the main constituent found in the plant and is used in oral and skin care. This review will summarize the phytochemistry, pharmacology, traditional uses and present dental uses of this plant.