Salivary Research Articles

Identification and culture of functional salivary gland ductal epithelial cells.

Sialadenitis is a prevalent salivary gland disease resulting in decreased salivary flow rate. To date, little is known about the exact changes and mechanism of ductal cells in sialadenitis. This study aims to establish an efficient method to identify and isolate ductal cells, thereby facilitating further research on this specific cell type. Immunofluorescence for cytokeratin 13 and cytokeratin 19 was conducted in salivary glands to confirm their specificity as ductal cell markers. The dissected ducts were assessed through PCR and Western blot of cytokeratin 19 and digested by dispase and collagenase. The functionality of the isolated ductal cells was determined by measuring intracellular calcium. Cytokeratin 19 and cytokeratin 13 were expressed in all segments of human ducts. Cytokeratin 19 was limited to ducts excluding granular convoluted tubules in rat and mouse. The purities of the obtained ductal cells were approximately 98% in humans and 93% in rats. Furthermore, intracellular free calcium increased with time and concentration of carbachol treatment. Cytokeratin 19 serves as a dependable marker for identifying ductal cells in salivary glands, except for granular convoluted tubules. Moreover, we have successfully developed an efficient method for isolating ductal cells from salivary glands.

Assessing parotid gland function through diffusion weighted MRI during and post-radiotherapy in head and neck cancer patients

ObjectivesThe purpose of this study was to evaluate the usefulness of apparent diffusion coefficient (ADC) variation in irradiated parotid glands during and after 3D conformal radiotherapy (3DCRT). Materials and methodsThis study enrolled 15 head and neck cancer (HNC) patients who were treated with 3DCRT underwent diffusion weighted imaging (DWI) at rest and after gustatory stimulation in three time points as follows: before, during (one day after receiving the mean dose 26 Gy) and 6 months after the end of radiotherapy (RT). Salivary Ejection Fraction (SEF) data was also obtained from salivary gland scintigraphy (SGS) at the same three time points as MRI. Mean ADC at rest and after stimulation (ADCr, ADCs) and SEF were extracted from parotid region for three time points. Finally, SEF and changes of ADC over time at rest and after stimulation and in each time point were compared. ResultsDifference between mean ADCr before RT and during RT was not significant (p = 0.12). The ADCr values were significantly higher after RT than before RT (p = 0.003) and during RT (after dose 26 Gy) (p = 0.001). The difference between ADCr and ADCs in each time point showed that there is significant difference between ADCr and ADCs before RT (p = 0.005). Difference between these parameters after RT was also significant (p = 0.05). Non-significant difference (p = 0.21) between ADCr and ADCs during RT was observed. The result of SEF also was significantly lower after RT than during (p = 0.008) and before RT (p = 0.001). ConclusionMean ADC values could be used as a surrogate marker to characterize the parotid function in different stages of RT.

The effect of smoking on amylase enzyme activity in saliva

Smoking has become a common habit in Indonesia. If done regularly, smoking is known to cause problems in the oral cavity. When smoking, cigarette smoke will enter the oral cavity and affect the activity of the salivary amylase enzyme. The journal search used in this review was conducted on several databases, such as PubMed, ScienceDirect, and Google with a range of years between 2013 and 2023. A total of 14 studies from various global populations were included in this review. Thirteen studies showed an association between smoking and alpha-amylase enzyme activity, while one study did not show an association between smoking and amylase enzyme. Of the 13 studies that showed an association, ten studies showed a negative association, while the remaining three studies showed a positive association. The effect of cigarettes on amylase activity in saliva may arise due to several factors, including the toxic effects of cigarettes on the salivary glands, mechanical and chemical stimulation of cigarettes on the mucosa, smoking period, oxidative stress index, and salivary pH.

Increased cell killing effect in neutron capture enhanced proton beam therapy

AbstractThermal neutrons generated in the body during proton beam therapy (PBT) can be used to cause boron neutron capture reactions and have recently been proposed as neutron capture enhanced PBT (NCEPBT). However, the cell killing effect of NCEPBT remains underexplored. Here, we show an increase in the cell killing effect of NCEPBT. Using Monte Carlo simulations, we showed that neutrons generated by proton beam irradiation are uniformly spread on tissue culture plates. Human salivary gland tumor cell line (HSG), human osteosarcoma cell line (MG63), human tongue squamous cell carcinoma cell line (SAS), and human malignant melanoma cell line (G-361) were irradiated with X-rays, proton beams, and proton beams with 10B-enriched boronophenylalanine (boron concentration of 20 and 80 ppm). The relative biological effectiveness (RBE) values of proton beams alone, proton beams with 20 ppm boron, and proton beams with 80 ppm boron for HSG, MG63, SAS, and G-361 were 1.02, 1.07, and 1.23; 1.01, 1.08, and 1.44; 1.05, 1.09, and 1.46; and 1.04, 1.13, and 1.63, respectively. NCEPBT with high boron concentration showed high RBE and a high sensitizing effect. Our results confirm an increase in the cell killing effect of NCEPBT, should aid in its clinical use, and warrant its further investigation.

Scanning Electron Microscopy of the Oropharyngeal Floor of Northern Bobwhite (Colinus virginianus, Linnaeus, 1758) Focusing on the Numerical and Regional Distribution of the Taste Buds

ABSTRACTThe architecture of the avian oropharynx corresponds to the feeding pattern. The present study aims to provide data on morphology, morphometry, and spatial distribution of taste buds in the oropharyngeal floor of the northern bobwhite (Colinus virginianus) using gross anatomy, morphometric, and scanning electron microscopy. The lower beak tip and tomia revealed the presence of variably‐sized dermal papillae. The prefrenular part of the oral floor had an interrupted median groove. Numerous taste buds and rostral submandibular salivary gland openings were detected alongside this groove. The tongue partially filled the oral floor. Its rostral end formed a median pointed process, the lingual nail. The dorsolateral aspects of the lingual apex and body showed hair‐like and rosette‐shaped filiform papillae of variable densities and sizes. However, these parts were devoid of taste buds. On the other hand, the surface mucosa of the lingual root presented numerous scale‐like cellular elevations and a large number of taste buds. Examination of cross sections in the lingual root suggested the presence of two groups of caudal lingual salivary glands, central and lateral. The laryngeal mound contained taste buds that protruded from the surface epithelium into the lumen. The mucosa of the pharyngeal floor contained many taste buds that appeared similar to those detected on the laryngeal mound, except for the large taste pores, excessive cellular porosity, and denser microvilli. The total proportions of the taste buds in the oral and pharyngeal floors were 34.1% and 65.81%, respectively. The proportions of taste buds were 34.1%, 57.26%, 4.27%, and 4.27% in the sublingual floor, lingual root, laryngeal mound, and pharyngeal wall, respectively. In conclusion, this study provides comprehensive data on the oropharynx of northern bobwhite and will help to understand mechanisms governing food gustation in the studied bird and other related birds.

A rare case of mucocele of the Blandin and Nuhn glands with review of literature

Oral mucocele is the most common benign minor salivary gland lesion of the oral cavity. Usually it occurs on lower lip, buccal mucosa and ventral surface of the tongue and rarely seen on ventral surface of the tongue, floor of mouth and palate. It is classified in two types of lesions, one is extravasation type which is more common and other is retention type. We present case report of a 37-year-old healthy male with complaint of swelling on ventral surface of the tongue, which was sessile, soft and fluctuant with smooth overlying mucosa with diagnosis of mucocele associated with Blandin and Nuhn gland. Management included marsupialization and drainage of thick mucus secretions. This article discusses on clinical features, diagnosis and management of mucoceles associated with Blandin and Nuhn gland.

Association between blood lead and periodontitis among American adults: a cross-sectional study of the national health and nutrition examination survey

BackgroundLead is persistent in the environment as a toxic substance and accumulates in the human body. Lead exposure has far-reaching harmful effects on all human systems and is widely recognized as a health and public health concern. Lead exposure poses a significant risk to oral health, as it destroys salivary glands and alveolar bone. It also induces oxidative stress which results in an immune response. Lead exposure appears to adversely affect periodontal tissues. Currently, the available evidence on the relationship between blood lead and periodontitis is insufficient and further research is necessary.ObjectiveIn this study, the objectives were to clarify the association between blood lead and periodontitis, as well as to explore potential dose-response relationships between blood lead exposure and periodontitis, as well as to determine appropriate thresholds for the effects of blood lead on periodontitis.MethodsWe conducted a cross-sectional study involving 8,550 participants with American adults aged 30 or older. blood Lead, periodontitis, age, sex, race, heart rate, education level, poverty index, marital status, body mass index, smoking status, alcohol drinking status, hypertension, diabetes and stroke. were collected from participants. Logistic regression, smooth curve fittingwere utilized to substantiate the research objectives.ResultsThere were a total of 8,550 participants of which 52.2% (4,467/8,550) had moderate/severe periodontitis. Compared with Q1 (0.15–0.93 ug/L), where blood lead levels were lower, the OR values for adjusted blood lead and periodontitis in Q2 (0.93–1.60 ug/L) and Q3 (1.60–61.29 ug/L) were 1.18 (95% CI: 1.12–1.25, P < 0.001) and 1.43 (95% CI: 1.34–1.52, P < 0.001). The association between blood lead levels and periodontitis exhibited a curve (non-linear, P < 0.001), with an inflection point of roughly 2.700 ug/L. The OR values for moderate/severe periodontitis in participants with blood lead levels <2.700ug/L was 1.318 (95% CI: 1.193–1.457, P < 0.001).ConclusionBlood lead levels are positively associated with periodontitis. Blood lead levels increased by five units and were associated with an increase in moderate and severe periodontitis risk by 36%. There is a curvilinear relationship between blood lead and periodontitis with a threshold effect and an inflection point of approximately 2.7 ug/L.

Submandibular Sialolithiasis Associated with Sialadenitis –A Case Report

Sialadenitis, an inflammation of major salivary glands is commonly associated with the obstruction of the salivary duct due to sialolith. The submandibular gland is particularly susceptible to sialolithiasis. This case report discusses a 33 year old female patient who reported with a swelling below the tongue/floor of the mouth associated with pain for a week A comprehensive investigation has indicated the presence of multiple sialoliths. The treatment involved surgical removal of the sialoliths, accompanied by antibiotic therapy. Therefore, this case report emphasizes the presence of multiple sialoliths in a female patient.

Diversity of hirudin and hirudin-like factor genes in the North-African medicinal leech, Hirudo troctina.

Medicinal leeches of the genus Hirudo inhabit large areas of the Palaearctic realm. The distribution range of Hirudo troctina includes the southern Iberian peninsula and the northwestern regions of Africa. H. troctina is used for medical purposes, but only very little is known about the components of its salivary gland secretion. Hirudins, bivalent inhibitors of thrombin, are probably the best known leech-derived bioactive factors. Hirudin-like factors (HLFs) represent another class of salivary gland components that share characteristic genetic and structural markers with hirudins. Hirudin is not a single entity but exists in at least four different variants. However, there are differences among the European members of the genus Hirudo with respect to the actual number of hirudin and HLF genes that are present within their genomes. Here, we describe the identification and molecular cloning of 11 genes that encode for putative hirudin and HLF variants in H. troctina. Three of the genes consist of exons and introns that originate from different "archetype" genes and are likely the result of recombination events. The diversity of hirudin and HLF genes in H. troctina surpasses that of all other European members of the genus Hirudo. The putative hirudin variants and representatives of the HLFs of H. troctina were expressed as recombinant proteins, purified and functionally characterized for their thrombin-inhibiting potencies. Phylogenetic analyses based on hirudin and HLF gene sequences of the leech genera Hirudo, Hirudinaria, and Whitmania supported the hypothesis that hirudins and HLFs diverged early in leech evolution.

Sclerosing mucoepidermoid carcinoma of salivary glands.

Sclerosing mucoepidermoid carcinoma (SMEC) of the salivary glands is a rare variant of low-grade mucoepidermoid carcinoma with scanty cellular atypia characterized by marked fibrosis/sclerosis and a rich inflammatory infiltrate. Herein, we report 25 unpublished cases of SMEC, two of them with prominent eosinophilia (2/25; 8%) and three with abundant IgG4-positive plasma cells (3/25; 12%). In our series of salivary SMEC, molecular analysis using fluorescence in situ hybridization (FISH) and/or next-generation sequencing (NGS) provided evidence of MAML2 gene rearrangement in 18 cases of the 21 analyzable cases tested (86%), while this gene locus was intact in 3 cases (14%). This study focuses on the diagnostic criteria of salivary SMEC given its challenge of abundant collagenous stroma, minimal residual neoplastic areas, and inconspicuous mucous cells. Follow-up data of our cases indicate that salivary SMECs have favorable outcomes. Molecular analysis for MAML2 gene rearrangement suggests that SMECs of salivary glands represent a rare variant of conventional low-grade MECs of salivary glands. In contrast, SMECs of the thyroid gland are genetically distinct from salivary-type thyroid MECs.

Salivary effector DcE1 suppresses plant defense and facilitates the successful feeding of Asian citrus psyllid, Diaphorina citri.

Piercing-sucking insects secrete diverse repertoires of effectors into their hosts to weaken host defenses and promote infestation. The Asian citrus psyllid, Diaphorina citri Kuwayama, is the most destructive insect pest in citrus orchards because of its role as a vector for the huanglongbing pathogen, Candidatus Liberibacter asiaticus (CLas). However, specific effector proteins and their functions in D. citri remain unclear. We demonstrate that DcE1, a salivary protein gene from D. citri, is predominantly expressed in the salivary gland tissues and is delivered into host plants during feeding. Transient expression in tobacco leaves revealed that DcE1 was subcellularly localized in the cytoplasm and plasma membrane, where it inhibited BAX- and INF1-induced cell death, suppressed callose deposition, and activated the salicylic acid pathway by upregulating the expression of endo-β-1,3-glucanase NtBGL2 and regulatory protein NtNPR1. Further, DcE1 knockdown by double-stranded RNA (dsRNA) injection decreased the survival rates of D. citri and interrupted D. citri phloem-feeding on host plants. These results indicate that DcE1 is a novel effector that promotes plant susceptibility and enables D. citri feeding. These findings enhance our understanding of D. citri-plant interactions and offer a potential new target gene for the development of citrus protection strategies. © 2024 Society of Chemical Industry.

Hydrogen peroxide disrupts the regulatory pathway of saliva secretion in two salivary acinar rat cell lines

BackgroundSecretion of saliva is controlled by autonomic nerve signals via regulation of Ca2+-dependent ion transport across acinar cell membranes. Oxidative stress may affect this process, leading to a decrease in saliva production. This study investigates elements of the Ca2+ regulatory pathway and their vulnerability to hydrogen peroxide-induced oxidative stress.MethodsRat parotid and submandibular salivary gland acinar cell lines were exposed to different hydrogen peroxide concentrations to simulate oxidative stress. Cell viability and intracellular reactive oxygen species were measured, mRNA levels were assessed via RT-qPCR, and protein expression was studied using western blot and immunofluorescence microscopy.ResultsElevated concentrations of hydrogen peroxide reduced cell viability and increased intracellular levels of reactive oxygen species and led to a decrease in cholinergic receptor muscarinic 3 and adrenoreceptor alpha 1A mRNA and protein levels in both cell lines. In parotid gland cells, both mRNA and protein levels of stromal interaction molecule 1 and Orai1 decreased with increasing concentrations of hydrogen peroxide. In contrast, in submandibular gland cells stromal interaction molecule 1 and Orai1 displayed differential mRNA and protein expression levels.ConclusionOur study revealed that hydrogen peroxide exposure alters rat parotid and submandibular acinar cells, increasing reactive oxygen species and reducing autonomic receptor expression. Differential mRNA and protein expression of stromal interaction molecule 1 and Orai1 highlight complex oxidative stress effects on Ca2⁺ signaling. Most likely these effects will be deleterious to salivary secretion, but some effects may be protective.

Trop2-Targeted Molecular Imaging in Solid Tumors: Current Advances and Future Outlook.

Trophoblast cell surface antigen 2 (Trop2), a transmembrane glycoprotein, plays a dual role in physiological and pathological processes. In healthy tissues, Trop2 facilitates development and orchestrates intracellular calcium signaling. However, its overexpression in numerous solid tumors shifts its function toward driving cell proliferation and metastasis, thus leading to a poor prognosis. The clinical relevance of Trop2 is underscored by its utility as both a biomarker for diagnostic imaging and a target for therapy. Notably, the U.S. Food and Drug Administration (FDA) has approved sacituzumab govitecan (SG), a novel Trop2-targeted agent, for treating triple-negative breast cancer (TNBC) and refractory urothelial cancer, highlighting the significance of Trop2 in clinical oncology. Molecular imaging, a powerful tool for visualizing and quantifying biological phenomena at the molecular and cellular levels, has emerged as a critical technique for studying Trop2. This approach encompasses various modalities, including optical imaging, positron emission tomography (PET), single photon emission computed tomography (SPECT), and targeted antibodies labeled with radioactive isotopes. Incorporating Trop2-targeted molecular imaging into clinical practice is vital for the early detection, prognostic assessment, and treatment planning of a broad spectrum of solid tumors. Our review captures the latest progress in Trop2-targeted molecular imaging, focusing on both diagnostic and therapeutic applications across diverse tumor types, including lung, breast, gastric, pancreatic, prostate, and cervical cancers, as well as salivary gland carcinomas. We critically evaluate the current state by examining the relevant applications, diagnostic accuracy, therapeutic efficacy, and inherent limitations. Finally, we analyze the challenges impeding widespread clinical application and offer insights into strategies for advancing the field, thereby guiding future research endeavors.

Australian Culex annulirostris mosquitoes are competent vectors for Japanese encephalitis virus genotype IV

Japanese encephalitis virus (JEV) is transmitted by Culex species of mosquitoes. In 2022, JEV belonging to a previously unrecognised lineage of genotype IV (GIV) caused a major outbreak of JE in South-eastern Australia, resulting in human cases and affecting piggeries. Cx. annulirostris has previously been implicated as the major vector of JEV in northern Australia where the virus has circulated since its first detection in 1995. Here, we showed that experimental infection of a laboratory colony of Australian Cx. annulirostris with the isolate JEV NSW/22 resulted in a 100% mosquito infection rate, with 87% of mosquito saliva samples testing positive by RT-qPCR at 14 days post-infection. Immunohistochemistry confirmed the presence of a replicating virus in the mosquito midgut and dissemination throughout the body, including the salivary glands. Our results also showed evidence of transovarial transmission of this virus; however, transstadial transmission from the eggs to the adult stage was not found. Comparison with an Indonesian isolate of GIV JEV and previous Australian isolates belonging to genotypes I and II showed that infection with JEV NSW/22 resulted in higher viral titres in the early stage of infection and higher proportions of mosquitoes with JEV-positive saliva, indicating a greater transmission potential compared to other isolates. This study provides compelling experimental evidence that Australian Cx. annulirostris is a highly efficient vector for the 2022 Australian JEV GIV outbreak strain.

Developmental studies of the sublingual and mandibular salivary glands in Japanese quails (Coturinx coturinx japonica).

The Japanese quail (Coturinx coturnix japonica) has a crucial role in the lives of humanity since the 12th century and continues to play main roles in our industry and scientific research. The advantages that the Japanese quail has, such as heavy egg production and high-quality meat with low cholesterol and fat contents, Moreover, the Japanese quail is easily managed, with high feeding conversion, low cost of investment, and high rate of returns. Salivary glands are a part of the lingual apparatus that secretes serios and mucous saliva. Whereas, the saliva secretions have different roles in the food variation, apprehension, and moisture of food bolus. The morphological and cytochemical analysis are done on 20 healthy Japanese quail embryos of 6th, 10th, 11th, and 13th days of incubation and 25 healthy quail chicks at hatching day old, 7th, 14th, 30th, and 60th days old. These samples are investigated histologically, histochemically, and scanned by electron microscopy serially. Our purpose of the study is to highlight the area of the oropharyngeal salivary glands and their role in food variation, as few studies spoke about that in Japanese quail. The primordia of the sublingual and mandibular salivary glands were noticed at the 6th and 10th days of the prehatching respectively as an epithelial bud. After hatching, both primordia were elongated and differentiated into secretory units. These glands were mucous polystomatic tubulo-alveolar paired glands, which were situated in the submucosa of the oropharyngeal floor (sublingual floor and paralingual grooves). The sublingual glands consisted of 3-5 lobes extended from the two Os ceratobranchial by their wide ends caudally, to beyond the median sulcus of the prefrenular part of the sublingual space rostrally. The taste buds were variable in size and position. The mandibular glands lay on the paralingual groove, which arose at the 10-day old embryo. The mandibular glands were located dorsomedial to the sublingual glands and extended longitudinally from the rostral border of the frenulum linguae to the caudal tips of the sublingual glands. The taste buds decreased in volume and number with advancing age. Overall, salivary glands increase in their alcianophilic activity of the secretions with advancing age, which indicates low PH within the secretory end pieces.

Warthin-like Papillary Thyroid Carcinoma: A Case Report and Review of the Literature.

Warthin-like subtype of papillary thyroid carcinoma (WLPTC) is a rare subtype of papillary thyroid carcinoma. This PTC subtype shows some pattern resemblance to Warthin tumors of the salivary gland, which confers its name but is a not genetically similar to salivary Warthin tumor, with the most frequent genetic change in WLPTC being BRAF p.V600E, the most frequent genetic driver of PTC. Warthin-like subtype of papillary thyroid carcinoma is found to have a favorable prognosis. We report a 44-year-old woman who presented with anterior neck swelling and elevated levels of antithyroid peroxidase antibodies. Fine-needle aspiration cytology showed features of papillary thyroid carcinoma and lymphocytic thyroiditis. On histopathological examination, thyroid follicular cells have oncocytic cytoplasm with infiltration of the tumor and fibrovascular cores by a dense lymphoplasmacytic infiltrate. Numerous multinucleated giant cells in the tumor are a notable feature. The final diagnosis is Warthin-like papillary thyroid carcinoma arising in association with chronic lymphocytic (Hashimoto) thyroiditis. Along with this case report, we perform a literature review of WLPTC between 2010 and 2024.

Positive results of treatment of adenocystic carcinoma of the parotid salivary gland with lenvatinib

Malignant tumors of the salivary glands account for 3–5 % of all head and neck cancers. Adenocystic carcinomas are malignant neoplasms that most often arise from large and small salivary glands. The disease is characterized by a progressive course with local and distant relapses.The article presents a clinical case of successful treatment with lenvatinib in suboptimal doses of adenocystic carcinoma of the parotid salivary gland.

Intraductal Papillary Mucinous Neoplasm: New Insights Into Its Origin and Nomenclatures.

Mucinous cells can be detected sporadically or may constitute the primary tumor component in salivary gland tumors, as observed in the intraductal papillary mucinous neoplasm (IPMN). This low-grade tumor is composed of mucinous columnar cells organized into papillary cystic structures. The present study aimed to compare the mucous cells in IPMN with mucous cells present in mucoepidermoid carcinoma (MEC) and papillary cystadenoma (PC). Immunohistochemistry analysis was carried out to compare the mucous cells in IPMN with the sporadic mucous cells in MEC (n = 4) and PC (n = 3). The results indicated that IPMN cells were positive for CK7, CK18, DOG1, and NKX3.1 and negative for CK14, SMA, and p63. The mucous cells in both MEC and PC were positive for CK7 and negative for CK18, SMA, DOG1, and NKX3.1. The positive expression of CK14 and p63 revealed the presence of basal cells both in PC, cystic areas of MEC, and normal mucous salivary glands. The immunohistochemical profile of IPMN closely resembles that of the mucous cells of the minor salivary glands yet differs from the mucous cells observed in MEC and PCX. This suggests that IPMN is probably derived from the transformation of secretory cells of the minor mucous salivary gland and has no rimming/basal cells. For this reason, we propose that this tumor is designated as mucous acinic cell carcinoma.

Multi-omics approaches define novel aphid effector candidates associated with virulence and avirulence phenotypes.

Compatibility between plant parasites and their hosts is genetically determined {Citation}both interacting organisms. For example, plants may carry resistance (R) genes or deploy chemical defences. Aphid saliva contains many proteins that are secreted into host tissues. Subsets of these proteins are predicted to act as effectors, either subverting or triggering host immunity. However, associating particular effectors with virulence or avirulence outcomes presents challenges due to the combinatorial complexity. Here we use defined aphid and host genetics to test for co-segregation of expressed aphid transcripts and proteins with virulent or avirulent phenotypes. We compared virulent and avirulent pea aphid parental genotypes, and their bulk segregant F1 progeny on Medicago truncatula genotypes carrying or lacking the RAP1 (Resistance to Acyrthosiphon pisum 1) resistance quantitative trait locus. Differential gene expression analysis of whole body and head samples, in combination with proteomics of saliva and salivary glands, enabled us to pinpoint proteins associated with virulence or avirulence phenotypes. There was relatively little impact of host genotype, whereas large numbers of transcripts and proteins were differentially expressed between parental aphids, likely a reflection of their classification as divergent biotypes within the pea aphid species complex. Many fewer transcripts intersected with the equivalent differential expression patterns in the bulked F1 progeny, providing an effective filter for removing genomic background effects. Overall, there were more upregulated genes detected in the F1 avirulent dataset compared with the virulent one. Some genes were differentially expressed both in the transcriptome and in the proteome datasets, with aminopeptidase N proteins being the most frequent differentially expressed family. In addition, a substantial proportion (27%) of salivary proteins lack annotations, suggesting that many novel functions remain to be discovered. Especially when combined with tightly controlled genetics of both insect and host plant, multi-omics approaches are powerful tools for revealing and filtering candidate lists down to plausible genes for further functional analysis as putative aphid effectors.

Efficacy of combined administration of Baekhogainsam-Tang and low-dose pilocarpine on frequent intractable xerostomia: Study protocol for a randomized controlled trial.

Intractable xerostomia is defined as the subjective perception of dry mouth and persistent salivary gland hypofunction. Pilocarpine is an approved salivary sialagogue that is frequently prescribed for the treatment of intractable xerostomia; however, it often exhibits more side effects at high-doses and limited effectiveness at low-doses. Baekhogainsam-Tang (BIT) is a common herbal formula used by patients complaining of sore throats and thirst. It seems that BIT can compensate for the insufficient effect of low-dose pilocarpine. However, no clinical trials have studied the efficacy of combined administration of BIT and low-dose pilocarpine for intractable xerostomia. We aim to assess the non-inferior efficacy and fewer side effects of combined administration of BIT and low-dose pilocarpine compared with the administration of high-dose pilocarpine. A randomized, open-label, parallel-group, multi-center trial will be conducted. A total of 120 patients with Sjogren's syndrome having an unstimulated salivary flow rate (SFR) ≤0.1 mL/min or who have undergone radiotherapy to the head and neck with an unstimulated SFR ≤0.25 mL/min will be recruited competitively. They will be randomly allocated to either the experimental or control groups. The experimental group will receive BIT herbal granules three times and pilocarpine (2.5-mg) four times daily; meanwhile, the control group will receive only 5-mg pilocarpine four times daily for 12 weeks. The primary outcome is unstimulated SFR after 12 weeks of treatment. Secondary outcomes are stimulated SFR after 12 weeks of medication, as well as differences and mean percentage changes in unstimulated and stimulated SFR, visual analog scale, salivary scintigraphy, and questionnaires for both oral symptoms and quality of life during the clinical trial. An independent T test or Mann-Whitney U test will be performed to compare values between the two groups. The Paired T test or Wilcoxon signed-rank test will be performed to compare intragroup continuous values. This trial will be significant evidence on the efficacy and safety of combined use of BIT and low-dose pilocarpine to treat intractable xerostomia. The Clinical Research Information Service of the Republic of Korea (ISRCTN, KCT0005982). Registered on 10 February 2021.