An ELISA to Detect Serum Antibodies to the Salivary Gland Toxin ofIxodes holocyclusNeumann in Dogs and Rodents

TheIxodes holocyclustick causes paralysis in up to 10,000 companion and domestic animals each year in Australia. Treatment requires the removal of the parasite and the administration of a commercial tick antiserum that is prepared from hyperimmune dogs. Each batch of this serum is initially tested for toxin-neutralising potency in a mouse bioassay that is expensive, time consuming, and subjective. With the aim of developing a rapidin vitroassay to replace the bioassay, we used a partially purified antigen prepared fromI. holocyclussalivary glands to develop an ELISA to detect toxin-reactive antibodies in hyperimmune dog sera. The optimised ELISA reliably detected antibodies reactive toI. holocyclussalivary gland antigens. Parallel testing of sera with a negative control antigen prepared from the salivary glands of the nontoxic tickRhipicephalus(Boophilus)microplusprovided further evidence that we were detecting toxin-specific antibodies in the assay. Using the ELISA, we could also detect antibodies induced in rats after experimental infestation withI. holocyclus. This assay shows promise as an alternative means of assessing the potency of batches of hyperimmune dog serum and to screen for toxin-reactive monoclonal antibodies produced from immunised rodents.

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Mesenchymal control over elongating and branching morphogenesis in salivary gland development

Development ◽

10.1242/dev.66.1.209 ◽

1981 ◽

Vol 66 (1) ◽

pp. 209-221

Author(s):

Hiroyuki Nogawa ◽

Takeo Mizuno

Keyword(s):

Salivary Gland ◽

Salivary Glands ◽

Submaxillary Gland ◽

Branching Morphogenesis ◽

Mouse Submaxillary Gland ◽

Gland Development

Recombination of the epithelium and mesenchyme between quail anterior submaxillary gland (elongating type) and quail anterior lingual or mouse submaxillary gland (branching type) was effected in vitro to clarify whether the elongating morphogenesis was directed by the epithelial or the mesenchymal component. Quail anterior submaxillary epithelium recombined with quail anterior lingual or mouse submaxillary mesenchyme came to branch. Conversely, quail anterior lingual or 12-day mouse submaxillary epithelium recombined with quail anterior submaxillary mesenchyme came to elongate, though the mesenchyme was less effective with 13-day mouse submaxillary epithelium. These results suggest that the elongating or branching morphogenesis of quail salivary glands is controlled by the mesenchyme.

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In vitro insecticidal effect of commercial fatty acids, ß-sitosterol and rutin against the sugarcane aphid, Melanaphis sacchari Zehntner (Hemiptera: Aphididae)

Journal of Food Protection ◽

10.4315/jfp-21-329 ◽

2022 ◽

Author(s):

Liliana Aguilar Marcelino ◽

Jesús Antonio Pineda Alegría ◽

David Osvaldo Salinas-Sánchez ◽

Víctor Manuel Hernández Velázquez ◽

Gonzalo Iván Silva Aguayo ◽

...

Keyword(s):

Fatty Acids ◽

Positive Control ◽

Negative Control ◽

Tween 20 ◽

Agricultural Pests ◽

Melanaphis Sacchari ◽

Insecticidal Effect ◽

Sugarcane Aphid ◽

Alternative Means

The sugarcane aphid, Melanaphis sacchari Zehntner (Hemiptera: Aphididae), is the main pest of sorghum, Sorghum bicolor L. Moench (Poaceae), in Mexico. To control this insect, farmers currently use synthetic chemical insecticides, which are toxic to humans and biodiversity. However, natural products are a promising potential source of safer alternative means to control different agricultural pests. The main objective of this study was to evaluate the insecticidal effect of contact by fumigation of pure molecules of four commercial fatty acids (palmitic, stearic, pentadecanoic and linoleic acids), the phytosterol ß -sitosterol, and the flavonoid rutin. The results showed that fatty acids were the most effective against M. sacchari ; the highest mortality rate (85%) was produced by linoleic acid and the LC 50 was 1,181 ppm, followed by stearic and palmitic acids with mortality percentages of 74 and 63%, respectively, at a concentration of 2,500 ppm at 72 h. The positive control, imidacloprid, had 100% mortality in 24 h and the tween 20 negative control exhibited 4% mortality in 72 h. Our results show that commercial fatty acids are effective against adults of M. sacchari , and can be considered an environmentally friendly alternative to the frequent use of synthetic chemical insecticides.

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Antibodies against MAEBL Ligand Domains M1 and M2 Inhibit Sporozoite Development In Vitro

Infection and Immunity ◽

10.1128/iai.72.6.3604-3608.2004 ◽

2004 ◽

Vol 72 (6) ◽

pp. 3604-3608 ◽

Cited By ~ 35

Author(s):

Peter Preiser ◽

Laurent Rénia ◽

Naresh Singh ◽

Bharath Balu ◽

William Jarra ◽

...

Keyword(s):

Salivary Gland ◽

Salivary Glands ◽

Vaccine Development ◽

Molecular Form ◽

Mammalian Host ◽

Phenotypic Differences ◽

Apical Membrane Antigen 1 ◽

Erythrocyte Binding ◽

Development In Vitro

ABSTRACT MAEBL is a type 1 membrane protein that is implicated in the merozoite invasion of erythrocytes and sporozoite invasion of mosquito salivary glands. This apical organelle protein is structurally similar to the ebl erythrocyte binding proteins, such as EBA-175, except that the tandem ligand domains of MAEBL are similar to part of the extracellular domain of apical membrane antigen 1 and not the Duffy binding-like domain. Although midgut and salivary gland sporozoites are morphologically similar, salivary gland sporozoites undergo a period of new gene expression after infecting the salivary glands, display distinct phenotypic differences, and are more infectious for the mammalian host. The objectives of this project were to determine the molecular form of MAEBL in the infectious salivary gland sporozoites and whether the ligand has a role in the sporozoite development to exoerythrocytic stages in hepatocytes. We determined that MAEBL is newly expressed in salivary gland sporozoites and in a form distinct from what is present in the midgut sporozoites or present in erythrocytic stages. Both ligand domains (M1 and M2) were expressed as part of a full-length membrane form of MAEBL in the salivary gland sporozoites in contrast to the other stages that retain only the M2 ligand domain as part of the membrane form of the protein. Antisera developed against the cysteine-rich regions of the extracellular portion of MAEBL inhibited sporozoite development to exoerythrocytic forms in vitro. Together these data indicate that MAEBL has a role in this third developmental stage in the life cycle of the malaria parasite. Thus, MAEBL is another target for pre-erythrocytic-stage vaccine development against malaria parasites.

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The influence of various factors on fluid secretion by in vitro salivary glands of ixodid Ticks

Journal of Experimental Biology ◽

10.1242/jeb.64.3.727 ◽

1976 ◽

Vol 64 (3) ◽

pp. 727-742

Author(s):

W. Kaufman

Keyword(s):

Salivary Gland ◽

Salivary Glands ◽

Culture Medium ◽

Dry Weight ◽

Fluid Secretion ◽

Ixodid Tick ◽

Ixodid Ticks ◽

Mammalian Tissue ◽

Stimulating Factor

1. Salivary glands of the female ixodid tick, Dermacentor andersoni, secrete fluid in vitro when bathed in a slightly modified version of the mammalian tissue culture medium ‘TC 199′. 2. Rate of salivation in vitro increases with progression of feeding, but there is no comparable increase in dry weight of the salivary glands during the early phase of engorgement. Engorged ticks secreted at only 25% the rate of 90–250 mg ticks, indicating that salivary gland degeneration has already begun in the very early post-engorgement stage. 3. A salivary gland stimulating factor can be detected in the nervous system but not in other tissues. 4. Male salivary glands secrete at only 1/20th the rate of female glands. Thus males probably do not use their salivary glands as osmoregulatory organs. 5. From the uniform lack of response to ACh and uniform response to DA in 7 ixodid tick species, it is suggested that the control of salivation is similar throughout the ixodid family.

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[177Lu]Lu-PSMA-617 Salivary Gland Uptake Characterized by Quantitative In Vitro Autoradiography

Pharmaceuticals ◽

10.3390/ph12010018 ◽

2019 ◽

Vol 12 (1) ◽

pp. 18 ◽

Cited By ~ 4

Author(s):

Roswitha Tönnesmann ◽

Philipp Meyer ◽

Matthias Eder ◽

Ann-Christin Baranski

Keyword(s):

Salivary Gland ◽

Salivary Glands ◽

Tumor Control ◽

Uptake Mechanism ◽

Future Design ◽

Gland Tissue ◽

Specific Uptake ◽

Therapy Strategies ◽

Higher Doses

Irradiation of salivary glands remains the main dose-limiting side effect of therapeutic PSMA-inhibitors, especially when using alpha emitters. Thus, further advances in radiopharmaceutical design and therapy strategies are needed to reduce salivary gland uptake, thereby allowing the administration of higher doses and potentially resulting in improved response rates and better tumor control. As the uptake mechanism remains unknown, this work investigates the salivary gland uptake of [177Lu]Lu-PSMA-617 by autoradiography studies on pig salivary gland tissue and on PSMA-overexpressing LNCaP cell membrane pellets. Displacement studies were performed with non-labeled PSMA-617 and 2-PMPA, respectively. The uptake of [177Lu]Lu-PSMA-617 in glandular areas was determined to be partly PSMA-specific, with a high non-specific uptake fraction. The study emphasizes that [177Lu]Lu-PSMA-617 accumulation in pig salivary glands can be attributed to a combination of both specific and non-specific uptake mechanisms. The observation is of high impact for future design of novel radiopharmaceuticals addressing the dose-limiting salivary gland irradiation of current alpha endoradiotherapy in prostate cancer.

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Development of a functional salivary gland tissue chip with potential for high-content drug screening

Communications Biology ◽

10.1038/s42003-021-01876-x ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Yuanhui Song ◽

Hitoshi Uchida ◽

Azmeer Sharipol ◽

Lindsay Piraino ◽

Jared A. Mereness ◽

...

Keyword(s):

Salivary Gland ◽

Salivary Glands ◽

Drug Screening ◽

Drug Testing ◽

In Vitro Models ◽

High Throughput Drug Screening ◽

Array Technology ◽

Gland Function ◽

Poly Ethylene

AbstractRadiation therapy for head and neck cancers causes salivary gland dysfunction leading to permanent xerostomia. Limited progress in the discovery of new therapeutic strategies is attributed to the lack of in vitro models that mimic salivary gland function and allow high-throughput drug screening. We address this limitation by combining engineered extracellular matrices with microbubble (MB) array technology to develop functional tissue mimetics for mouse and human salivary glands. We demonstrate that mouse and human salivary tissues encapsulated within matrix metalloproteinase-degradable poly(ethylene glycol) hydrogels formed in MB arrays are viable, express key salivary gland markers, and exhibit polarized localization of functional proteins. The salivary gland mimetics (SGm) respond to calcium signaling agonists and secrete salivary proteins. SGm were then used to evaluate radiosensitivity and mitigation of radiation damage using a radioprotective compound. Altogether, SGm exhibit phenotypic and functional parameters of salivary glands, and provide an enabling technology for high-content/throughput drug testing.

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Laminin-1 Peptides Conjugated to Fibrin Hydrogels Promote Salivary Gland Regeneration in Irradiated Mouse Submandibular Glands

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2021.729180 ◽

2021 ◽

Vol 9 ◽

Author(s):

Kihoon Nam ◽

Harim T. dos Santos ◽

Frank Maslow ◽

Bryan G. Trump ◽

Pedro Lei ◽

...

Keyword(s):

Radiation Therapy ◽

Salivary Gland ◽

Salivary Glands ◽

Tissue Regeneration ◽

Secretory Function ◽

Gland Morphogenesis ◽

Salivary Gland Regeneration ◽

Laminin 1

Previous studies demonstrated that salivary gland morphogenesis and differentiation are enhanced by modification of fibrin hydrogels chemically conjugated to Laminin-1 peptides. Specifically, Laminin-1 peptides (A99: CGGALRGDN-amide and YIGSR: CGGADPGYIGSRGAA-amide) chemically conjugated to fibrin promoted formation of newly organized salivary epithelium both in vitro (e.g., using organoids) and in vivo (e.g., in a wounded mouse model). While these studies were successful, the model’s usefulness for inducing regenerative patterns after radiation therapy remains unknown. Therefore, the goal of the current study was to determine whether transdermal injection with the Laminin-1 peptides A99 and YIGSR chemically conjugated to fibrin hydrogels promotes tissue regeneration in irradiated salivary glands. Results indicate that A99 and YIGSR chemically conjugated to fibrin hydrogels promote formation of functional salivary tissue when transdermally injected to irradiated salivary glands. In contrast, when left untreated, irradiated salivary glands display a loss in structure and functionality. Together, these studies indicate that fibrin hydrogel-based implantable scaffolds containing Laminin-1 peptides promote secretory function of irradiated salivary glands.

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Sox9+ Cell is Required for Salivary Gland Regeneration after Radiation Damage via Wnt/β-Catenin Pathway

10.21203/rs.3.rs-535738/v1 ◽

2021 ◽

Author(s):

Xiuyun Xu ◽

Xiong Gan ◽

Ming Zhang ◽

Jiaxiang Xie ◽

Shuang Chen ◽

...

Keyword(s):

Salivary Gland ◽

Salivary Glands ◽

Cell Lineage ◽

Term Treatment ◽

Sequencing Data ◽

Long Term Treatment ◽

Radiation Induced ◽

Gland Function ◽

Salivary Gland Regeneration

Abstract Background: Radiotherapy for head and neck cancer can cause serious side effects, including severe damage to the salivary glands, resulting in symptoms such as xerostomia, dental caries, oral infectious and so on. Due to lack of long-term treatment for the symptoms of saliva barren, current research has focused on finding endogenous stem cells that can differentiate into various cell lineage to replace lost tissue and restore function. Results: In our study, we identified Sox9+ cells can differentiate into various salivary epithelial cell lineages under homeostatic conditions. After ablating Sox9+ cells, the salivary glands of irradiated mice showed more severe phenotypes and reduced proliferative capacity. Analysis of online single cell RNA-sequencing data revealed enrichment of the Wnt/β-catenin pathway in Sox9+ cell population. Furthermore, treatment of Wnt/β-catenin inhibitor to irradiated mice inhibited the regenerative capability of Sox9+ cells. Finally, we showed that Sox9+ cells were able to form organoids in vitro and transplanting these organoids into salivary glands after radiation restored part of salivary gland function. Conclusions: In short, our research indicated that regenerative therapy targeting Sox9+ cells is a promising method to solve the radiation induced salivary gland injury.

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The murine cytomegalovirus MCK-2 facilitates in vivo infection transfer from dendritic cells to salivary gland acinar cells.

Journal of Virology ◽

10.1128/jvi.00693-21 ◽

2021 ◽

Author(s):

Jiawei Ma ◽

Kimberley Bruce ◽

Philip G. Stevenson ◽

Helen E. Farrell

Keyword(s):

Dendritic Cells ◽

Salivary Gland ◽

Epithelial Cells ◽

Salivary Glands ◽

Immune Evasion ◽

Acinar Cells ◽

Lung Infection ◽

Human Cmv

The cytomegaloviruses (CMVs) spread systemically via myeloid cells and demonstrate a broad tissue tropism. Human CMV (HCMV) UL128 encodes a component of the virion pentameric complex (PC) that is important for entry into epithelial cells and cell-cell spread in vitro . It possesses N-terminal amino acid sequences similar to C-C chemokines. While the species-specificity of HCMV precludes confirmation of UL128 function in vivo , UL128-like counterparts in experimental animals have demonstrated a role for salivary gland infection. How they achieve this has not been defined, although effects on monocyte tropism and immune evasion have been proposed. By tracking infected cells following lung infection, we show that although the UL128-like protein in mouse CMV (MCMV; designated MCK-2), facilitated entry into lung macrophages, it was dispensable for subsequent viremia mediated by CD11c + dendritic cells (DC) and extravasation to the salivary glands. Notably, MCK-2 was important for transfer of MCMV infection from DC to salivary gland acinar epithelial cells. Acinar cell infection of MCMVs deleted of MCK-2 was not rescued by T-cell depletion, arguing against an immune evasion mechanism for MCK-2 in the salivary glands. In contrast to lung infection, peritoneal MCMV inoculation yields a mixed monocyte/DC viremia. In this setting, MCK-2 again promoted DC-dependent infection of salivary gland acinar cells, but it was not required for monocyte-dependent spread to the lung. Thus, the action of MCK-2 in MCMV spread was specific to DC-acinar cell interaction. IMPORTANCE Cytomegaloviruses (CMVs) establish myeloid cell-associated viremias and persistent shedding from the salivary glands. In vitro studies with human CMV (HCMV) have implicated HCMV UL128 in epithelial tropism, but its role in vivo is unknown. Here we analysed how a murine CMV (CMV) protein with similar physical properties - designated MCK-2 - contributes to host colonization. We demonstrate that MCK-2 is dispensable for the initial systemic spread from primary infection sites, but within the salivary gland facilitates transfer of infection from dendritic cells (DC) to epithelial acinar cells. Virus transfer from extravasated monocytes to the lungs did not require MCK-2, indicating a tissue-specific effect. These results provide new information about how persistent viral tropism determinants operate in vivo .

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Regenerating Salivary Glands in the Microenvironment of Induced Pluripotent Stem Cells

BioMed Research International ◽

10.1155/2015/293570 ◽

2015 ◽

Vol 2015 ◽

pp. 1-11 ◽

Cited By ~ 19

Author(s):

Hitomi Ono ◽

Aya Obana ◽

Yu Usami ◽

Manabu Sakai ◽

Kanji Nohara ◽

...

Keyword(s):

Stem Cells ◽

Salivary Gland ◽

Salivary Glands ◽

Ips Cells ◽

Initial Attempt ◽

Cell Niche ◽

Salivary Gland Regeneration ◽

Induced Pluripotent

This report describes our initial attempt to regenerate salivary glands using induced pluripotent stem (iPS) cellsin vivoandin vitro. Glandular tissues that were similar to the adult submandibular glands (SMGs) and sublingual glands could be partially produced by the transplantation of iPS cells into mouse salivary glands. However, the tumorigenicity of iPS cells has not been resolved yet. It is well known that stem cells affect their microenvironment, known as a stem cell niche. We focused on the niche and the interaction between iPS cells and salivary gland cells in our study on salivary gland regeneration. Coculture of embryonic SMG cells and iPS cells have better-developed epithelial structures and fewer undifferentiated specific markers than monoculture of embryonic SMG cellsin vitro. These results suggest that iPS cells have a potential ability to accelerate differentiation for salivary gland development and regeneration.

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