Adult Gland Research Articles

Loss of the disease-associated glycosyltransferase Galnt3 alters Muc10 glycosylation and the composition of the oral microbiome

Loss of the disease-associated glycosyltransferase Galnt3 alters Muc10 glycosylation and the composition of the oral microbiome

Circadian regulation and molecular role of the Bsx homeobox gene in the adult pineal gland.

The pineal gland is a neuroendocrine organ responsible for production of the nocturnal hormone melatonin. A specific set of homeobox gene-encoded transcription factors govern pineal development, and some are expressed in adulthood. The brain-specific homeobox gene (Bsx) falls into both categories. We here examined regulation and function of Bsx in the mature pineal gland of the rat. We report that Bsx is expressed from prenatal stages into adulthood, where Bsx transcripts are localized in the melatonin-synthesizing pinealocytes, as revealed by RNAscope in situ hybridization. Bsx transcripts were also detected in the adult human pineal gland. In the rat pineal gland, Bsx was found to exhibit a 10-fold circadian rhythm with a peak at night. By combining in vivo adrenergic stimulation and surgical denervation of the gland in the rat with in vitro stimulation and transcriptional inhibition in cultured pinealocytes, we show that rhythmic expression of Bsx is controlled at the transcriptional level by the sympathetic neural input to the gland acting via adrenergic stimulation with cyclic AMP as a second messenger. siRNA-mediated knockdown (>80% reduction) in pinealocyte cultures revealed Bsx to be a negative regulator of other pineal homeobox genes, including paired box 4 (Pax4), but no effect on genes encoding melatonin-synthesizing enzymes was detected. RNA sequencing analysis performed on siRNA-treated pinealocytes further revealed that downstream target genes of Bsx are mainly involved in developmental processes. Thus, rhythmic Bsx expression seems to govern other developmental regulators in the mature pineal gland.

Dynamic architectural interplay between leucocytes and mammary epithelial cells.

The adult mammary gland undergoes dynamic changes during puberty and the postnatal developmental cycle. The mammary epithelium is composed of a bilayer of outer basal, or myoepithelial, cells and inner luminal cells, the latter lineage giving rise to the milk‐producing alveolar cells during pregnancy. These luminal alveolar cells undergo Stat3‐mediated programmed cell death following the cessation of lactation. It is established that immune cells in the microenvironment of the gland have a role to play both in the ductal outgrowth during puberty and in the removal of dead cells and remodelling of the stroma during the process of postlactational regression. However, most studies have focussed on the role of the stromal immune cell compartment or have quantified immune cell populations in tissue extracts. Our recent development of protocols for deep imaging of the mammary gland in three dimensions (3D) has enabled the architectural relationship between immune cells and the epithelium to be examined in detail, and we have discovered a surprisingly dynamic relationship between the basal epithelium and leucocytes. Furthermore, we have observed morphological changes in the myoepithelial cells, as involution progresses, which were not revealed by previous work in 2D tissue sections and whole tissue. This dynamic architecture suggests a role for myoepithelial cells in the orderly progression of involution. We conclude that deep imaging of mammary gland and other tissues is essential for analysing complex interactions between cellular compartments.

Exogenous ERα Expression in the Mammary Epithelium Decreases Over Time and Does Not Contribute to p53-Deficient Mammary Tumor Formation in Mice.

Approximately 75% of all breast cancers express the nuclear hormone receptor estrogen receptor α (ERα). However, the majority of mammary tumors from genetically engineered mouse models (GEMMs) are ERα-negative. To model ERα-positive breast cancer in mice, we exogenously introduced expression of mouse and human ERα in an existing GEMM of p53-deficient breast cancer. After initial ERα expression during mammary gland development, expression was reduced or lost in adult glands and p53-deficient mammary tumors. Chromatin immunoprecipitation (ChIP)-sequencing analysis of primary mouse mammary epithelial cells (MMECs) derived from these models, in which expression of the ERα constructs was induced in vitro, confirmed interaction of ERα with the DNA. In human breast and endometrial cancer, and also in healthy breast tissue, DNA binding of ERα is facilitated by the pioneer factor FOXA1. Surprisingly, the ERα binding sites identified in primary MMECs, but also in mouse mammary gland and uterus, showed an high enrichment of ERE motifs, but were devoid of Forkhead motifs. Furthermore, exogenous introduction of FOXA1 and GATA3 in ERα-expressing MMECs was not sufficient to promote ERα-responsiveness of these cells. Together, this suggests that species-specific differences in pioneer factor usage between mouse and human are dictated by the DNA sequence, resulting in ERα-dependencies in mice that are not FOXA1 driven. These species-specific differences in ERα-biology may limit the utility of mice for in vivo modeling of ERα-positive breast cancer.

Genomic analysis of the tryptome reveals molecular mechanisms of gland cell evolution

BackgroundUnderstanding the drivers of morphological diversity is a persistent challenge in evolutionary biology. Here, we investigate functional diversification of secretory cells in the sea anemone Nematostella vectensis to understand the mechanisms promoting cellular specialization across animals.ResultsWe demonstrate regionalized expression of gland cell subtypes in the internal ectoderm of N. vectensis and show that adult gland cell identity is acquired very early in development. A phylogenetic survey of trypsins across animals suggests that this gene family has undergone numerous expansions. We reveal unexpected diversity in trypsin protein structure and show that trypsin diversity arose through independent acquisitions of non-trypsin domains. Finally, we show that trypsin diversification in N. vectensis was effected through a combination of tandem duplication, exon shuffling, and retrotransposition.ConclusionsTogether, these results reveal the numerous evolutionary mechanisms that drove trypsin duplication and divergence during the morphological specialization of cell types and suggest that the secretory cell phenotype is highly adaptable as a vehicle for novel secretory products.

Candidates for chemosensory genes identified in the Chinese citrus fly, Bactrocera minax, through a transcriptomic analysis

BackgroundThe males of many Bactrocera species (Diptera: Tephritidae) respond strongly to plant-derived chemicals (male lures) and can be divided into cue lure/raspberry ketone (CL/RK) responders, methyl eugenol (ME) responders and non-responders. Representing a non-responders, Bactrocera minax display unique olfactory sensory characteristics compared with other Bactrocera species. The chemical senses of insects mediate behaviors that are associated with survival and reproduction. Here, we report the generation of transcriptomes from antennae and the rectal glands of both male and female adults of B. minax using Illumina sequencing technology, and annotated gene families potentially responsible for chemosensory.ResultsWe developed four transcriptomes from different tissues of B. minax and identified a set of candidate genes potentially responsible for chemosensory by analyzing the transcriptomic data. The candidates included 40 unigenes coding for odorant receptors (ORs), 30 for ionotropic receptors (IRs), 17 for gustatory receptors (GRs), three for sensory neuron membrane proteins (SNMPs), 33 for odorant-binding proteins (OBPs), four for chemosensory proteins (CSPs). Sex- and tissue-specific expression profiles for candidate chemosensory genes were analyzed via transcriptomic data analyses, and expression profiles of all ORs and antennal IRs were investigated by real-time quantitative PCR (RT-qPCR). Phylogenetic analyses were also conducted on gene families and paralogs from other insect species together.ConclusionsA large number of chemosensory genes were identified from transcriptomic data. Identification of these candidate genes and their expression profiles in various tissues provide useful information for future studies towards revealing their function in B. minax.

A Vaccinomics Approach for the Identification of Tick Protective Antigens for the Control of Ixodes ricinus and Dermacentor reticulatus Infestations in Companion Animals.

Ticks and tick-borne pathogens affect health and welfare of companion animals worldwide, and some human tick-borne diseases are associated with exposure to domestic animals. Vaccines are the most environmentally friendly alternative to acaracides for the control of tick infestations, and to reduce the risk for tick-borne diseases affecting human and animal health. However, vaccines have not been developed or successfully implemented for most vector-borne diseases. The main limitation for the development of effective vaccines is the identification of protective antigens. To address this limitation, in this study we used an experimental approach combining vaccinomics based on transcriptomics and proteomics data with vaccination trials for the identification of tick protective antigens. The study was focused on Ixodes ricinus and Dermacentor reticulatus that infest humans, companion animals and other domestic and wild animals, and transmit disease-causing pathogens. Tick larvae and adult salivary glands were selected for analysis to target tick organs and developmental stages playing a key role during tick life cycle and pathogen infection and transmission. Two I. ricinus (heme lipoprotein and uncharacterized secreted protein) and five D. reticulatus (glypican-like protein, secreted protein involved in homophilic cell adhesion, sulfate/anion exchanger, signal peptidase complex subunit 3, and uncharacterized secreted protein) proteins were identified as the most effective protective antigens based on the criteria of vaccine E > 80%. The putative function of selected protective antigens, which are involved in different biological processes, resulted in vaccines affecting multiple tick developmental stages. These results suggested that the combination of some of these antigens might be considered to increase vaccine efficacy through antigen synergy for the control of tick infestations and potentially affecting pathogen infection and transmission. These antigens were proposed for commercial vaccine development for the control of tick infestations in companion animals, and potentially in other hosts for these tick species.

Inhibition of epidermal growth factor receptor stimulates prolactin expression in primary culture of the mouse pituitary gland.

The roles of epidermal growth factor (EGF) in the regulation of prolactin (PRL) gene expression in the normal pituitary gland remain poorly understood. In the present study, the effects of EGF and an inhibitor of the EGF receptor, erlotinib, on PRL gene expression were examined both in the pituitary tumour cell line GH3 and in a primary culture of the mouse pituitary gland under similar experimental conditions. The results showed that EGF stimulated PRL expression in GH3 cells, but not in normal cells. Erlotinib was found to counteract EGF in GH3 cells inhibiting the PRL expression enhanced by EGF. By contrast, erlotinib induced an elevation in the PRL mRNA levels in the primary culture of the adult pituitary gland and the initiation of PRL production in the culture of the foetal pituitary gland in which PRL production had not yet occurred. Western blot analyses showed that EGF induced and erlotinib inhibited the activation of extracellular regulated protein kinase equally in GH3 and normal cells. These results suggest that the consequences of EGF receptor activation in normal PRL cells contradict those in adenomatous PRL cells.

Ultrasonographic and elastographic biometry in adult major salivary glands: a preliminary case-control report

Specifications about the size and stiffness of healthy salivary glands with ultrasound (US) are not available for Asian people. Using a Toshiba Apolio 500 US platform, we determined the size (including anterior-posterior median length, median paramandibular depth dimension, and cranio-caudal height) and hardness of 100 healthy submandibular and parotid glands in volunteers without a history of disease affecting the salivary glands or post-radiation, and compared the dimensions to those of 36 parotid glands and 37 submandibular glands in post-irradiated patients. The dimensions of the parotid and submandibular glands were significantly correlated with body weight. However, the dimension of the parotid glands was not significantly correlated with that of patients with prior radiation; the shear wave velocity (SWV) significantly increased (1.99 m/s versus 2.43 m/s, p-value < 0.01). The dimension of the submandibular glands was significantly correlated with prior radiation, where the SWV also significantly increased (2.32 m/s versus 2.50 m/s, p-values < 0.01). We find that US is a useful tool for assessment of the reference dimensions and hardness of major salivary glands that may be altered by irradiation.

Concise Review: A Critical Evaluation of Criteria Used to Define Salivary Gland Stem Cells.

In the effort to develop cell-based therapies to treat salivary gland dysfunction, many different populations of cells in the adult salivary glands have been proposed as stem cells. These cell populations vary, depending on the assay used, and are often nonoverlapping, leading to the conclusion that salivary glands harbor multiple stem cells. The goal of this review is to critically appraise the assays and properties used to identify stem cells in the adult salivary gland, and to consider the caveats of each. Re-evaluation of the defining criteria may help to reconcile the many potential stem cell populations described in the salivary gland, in order to increase comparability between studies and build consensus in the field. Stem Cells 2019;37:1144-1150.

Volatile Compounds Emitted by the Stink Bug Antiteuchus innocens (Hemiptera: Pentatomidae)

In this study, volatile compounds emitted by disturbed nymphs and adults of the stink bug Antiteuchus innocens were identified by gas chromatography-mass spectrometry as follows: (E)-hexenal, α-pinene, (E)-2-heptenal, n-decane, (E)-2-octenal, n-undecane, n-dodecane, and n-tridecane. The major components were (E)-2-hexenal, (E)-2-heptenal, and n-undecane. The same compounds, except α-pinene and (E)-2-octenal, were found in the dorsal glands and metathoracic glands of nymphs and adults, respectively. Extracts of exocrine glands did not elicit antennal responses of the adults, or affect the behavior of fifth instar nymphs, which suggests that these compounds are likely involved in the A. innocens defense rather than functioning as an alarm pheromone. Sumario En este estudio, los compuestos volatiles emitidos por las ninfas y adultos molestados de la chinche apestosa Antiteuchus innocens fueron identificados por cromatografia de gases-espectrometria de masas como (E)-hexenal, α-pineno, (E)-2-heptenal, n-decano, (E)-2-octenal, n-undecano, n-dodecano, and n-tridecano. Los compuestos mayoritarios de la mezcla fueron el (E)-2-hexenal, (E)-2-heptenal, and n-undecano. Los mismos compuestos, excepto el α-pineno and (E)-2-octenal, fueron encontrados en las glandulas dorsales y metatoracicas de ninfas y adultos, respectivamente. Extractos de las glandulas exocrinas no evocaron una respuesta antenal de adultos, ni afectaron el comportamiento de ninfas de quinto instar, lo que sugiere que estos compuestos estan probablemente involucrados en funciones de defensas mas que como una feromona de alarma. Key Words: Pentatomidae; Discocephalinae; metathoracic glands; dorsal glands; volatiles View this article in BioOne https://doi.org/10.1653/024.102.0223

Neurturin-containing laminin matrices support innervated branching epithelium from adult epithelial salispheres

Cell transplantation of autologous adult biopsies, grown ex vivo as epithelial organoids or expanded as spheroids, are proposed treatments to regenerate damaged branching organs. However, it is not clear whether transplantation of adult organoids or spheroids alone is sufficient to initiate a fetal-like program of branching morphogenesis in which coordinated branching of multiple cell types including nerves, mesenchyme and blood vessels occurs. Yet this is an essential concept for the regeneration of branching organs such as lung, pancreas, and lacrimal and salivary glands. Here, we used factors identified from fetal organogenesis to maintain and expand adult murine and human epithelial salivary gland progenitors in non-adherent spheroid cultures, called salispheres. These factors stimulated critical developmental pathways, and increased expression of epithelial progenitor markers such as Keratin5, Keratin14, FGFR2b and KIT. Moreover, physical recombination of adult salispheres in a laminin-111 extracellular matrix with fetal salivary mesenchyme, containing endothelial and neuronal cells, only induced branching morphogenesis when neurturin, a neurotrophic factor, was added to the matrix. Neurturin was essential to improve neuronal survival, axon outgrowth, innervation of the salispheres, and resulted in the formation of branching structures with a proximal-distal axis that mimicked fetal branching morphogenesis, thus recapitulating organogenesis. Epithelial progenitors were also maintained, and developmental differentiation programs were initiated, showing that the fetal microenvironment provides a template for adult epithelial progenitors to initiate branching and differentiation. Further delineation of secreted and physical cues from the fetal niche will be useful to develop novel regenerative therapies that instruct adult salispheres to resume a developmental-like program in vitro and to regenerate branching organs in vivo.

Sox9 regulates the luminal stem/progenitor cell properties of salivary glands

Exocrine glands share a common morphology consisting of ductal, acinar, and basal/myoepithelial cells, but their functions and mechanisms of homeostasis differ among tissues. Salivary glands are an example of exocrine glands, and they have been reported to contain multipotent stem cells that differentiate into other tissues. In this study, we purified the salivary gland stem/progenitor cells of adult mouse salivary glands using the cell surface marker CD133 by flow cytometry. CD133+ cells possessed stem cell capacity, and the transplantation of CD133+ cells into the submandibular gland reconstituted gland structures, including functional acinar. CD133+ cells were sparsely distributed in the intercalated and exocrine ducts and expressed Sox9 at higher levels than CD133– cells. Moreover, we demonstrated that Sox9 was required for the stem cell properties CD133+ cells, including colony and sphere formation. Thus, the Sox9-related signaling may control the regeneration salivary glands.

OR26-3 ERK1/2-RSK2 Maintains Estrogen Homeostasis in the Adult Mammary Gland

In response to estrogens, estrogen receptor alpha (ERα) is degraded by the 26S proteasome; however, despite the continued presence of estrogen prior to menopause, ERα protein levels are maintained by an unidentified mechanism. We find that the Ser/Thr kinase, RSK2, negatively regulates ERK1/2-dependent ERα degradation in the mammary gland to maintain ERα protein levels in cycling adult female mice. In RSK2 knockout (RSK2-KO) mice, ERα protein levels are decreased ~70% in the adult mammary gland compared to the control. Mammary gland regeneration experiments demonstrated that regulation of ERα protein levels by RSK2 is intrinsic to the mammary epithelium. In juvenile animals the ERK1/2-RSK2 pathway is not active and consistent with that observation the ERα protein levels are similar in juvenile wild type and RSK2-KO mice. In adults, the levels of active ERK1/2 increase ~25-fold relative to juvenile mammary glands. Activation of ERK1/2 in the adult is restricted to the estrus phase of the estrous cycle and activation of ERK1/2 is estrogen-dependent. Oophorectomy or inhibition of the 26S proteasome restored ERα protein levels in RSK2-KO mice. Loss of RSK2 results in activation of ERK1/2 throughout the estrous cycle and results in an enrichment for estrogen-responsive genes in the RSK2-KO ER+ mammary epithelial cells relative to wild type. These data support a model in which the unrestricted activation of ERK1/2 in RSK2-KO mice drives inappropriate transcription-coupled proteolysis of ERα which leads to an increase in DNA-double stranded breaks. Bioinformatic analysis demonstrated a correlation between decreased RSK2 expression and increased expression of estrogen-regulated gene in normal breast tissue from women taking oral contraceptives. Furthermore, the RSK2-KO transcriptome signature identified a subset of ER+ breast cancer patients with low RSK2 expression. These findings establish RSK2 as a regulator of estrogen homeostasis in the pre-menopausal adult and suggest a possible mechanism for the increased cancer risk in women taking exogenous estrogens. Funding source: NIH

The differential secretion of FSH and LH: regulation through genes, feedback and packaging

While the role of oestradiol and progesterone in the control of GnRH pulsatile secretion and generation of the preovulatory GnRH surge to induce release of the LH surge has been fully investigated, less attention has been given to changes in the pituitary gland that may sensitize gonadotrophs to switch from pulsatile release to surge release of LH, in particular. Furthermore, in the follicular phase while pulsatile secretion of LH is maximal, FSH secretion is reduced, yet both hormones are produced by the same gonadotrophs. The mechanisms whereby this differential release can occur are still unclear. The main regulator of FSH secretion is through the negative feedback effects of oestradiol and inhibin, which directly affect FSHbeta mRNA content and subsequent synthesis of FSH. FSH is then released predominantly via a constitutive pathway and the amount released is closely related to the rate of synthesis. In contrast, while basal LH secretion occurs via a constitutive pathway, the principal release of LH through pulsatile secretion is through the regulated pathway with GnRH stimulating the release of pre-synthesized LH contained in storage granules without significant changes in LHbeta mRNA. Secretogranin II (SgII) is associated with LH in these electron-dense storage granules and LH-SgII granules appear to be the principal form of granule released in response to GnRH through the regulated pathway. At the time of the preovulatory LH surge, granule movement to the gonadotrope cell membrane abutting a capillary, polarization, appears to play an important part in the priming mechanism for release of LH during the preovulatory LH surge in response to the GnRH surge. As there appears to be limited or no gonadotroph cell division in the adult pituitary gland, each gonadotroph passes through this synthesis and secretion pathway repeatedly through successive oestrous cycles. Packaging of LH and FSH into different secretory granules within the same cell is thus pivotal for the differential secretion of these gonadotrophins.

Human Submandibular Salivary Gland Morphology Associated to Transcription and Growth Factors from Fetus to Aging

Saliva carries out several functions in the digestive system like the protection of the oral cavity and the beginning of carbohydrate digestion. This liquid is mainly produced by the submandibular gland as it is responsible for 60% of total saliva flow. This gland is characterized as acinar tubule composed, and during the fetal period, it develops in five stages, which are molded through high proliferative indices of the components. However, the aging process changes the morphology and function of this gland.Transcription factors such as Nanog and Signal transducer and activator of transcription 3 (Stat‐3) regulate the process of proliferation, ckit and musashi‐1 induce cell proliferation while epidermal growth factor (EGF) is secreted by the submandibular gland and it has combined action to the Stat‐3. Considering the importance of these transcription factors and the role of EGF, the present study evaluated immunohistochemical expression of EGF, Nanog, Ckit, Musashi‐1 and Stat‐3 in submandibular glands of human fetuses between 12 and 18 gestational weeks, in adults (40 to 55 years old) and senile (over 85 years old). It was also evaluated quantitatively the morphological changes in components during development using Image J and statistical softwares. This study was approved by the Research Ethical Committee of UNIFESP, research number 618131.The present study showed significant expression of all the transcription factors in the acinar system as in the ductal system during the stages of development. Also showed differences in the number of components between the adult and senile glands. Considering these results, we reinforce that the morphological changes observed in the submandibular gland is in consonance with the levels of the studied markers and the proliferation pattern during aging.Support or Funding InformationThis study was financed in part by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior ‐ Brasil (CAPES) Finance code 001 and also by FAPESP process number 2015/03967‐5.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Sexual dimorphisms in the transcriptomes of murine salivary glands.

Transcriptional profiling identified 933 sexually dimorphic genes out of the 14 371 protein‐coding genes expressed in the three major murine salivary glands: parotid, sublingual, and submandibular. Most (89%) sex‐specific genes were enriched in a single gland, while only 0.5% of the sexually dimorphic genes were enriched in all glands. The sublingual gland displayed a strong male sex bias (94% of sex‐enriched genes), while a sex preference was not obvious in the parotid or submandibular glands. A subset of transcription factor genes was correlated with the expression of gland‐specific, sex‐enriched genes. Higher expression of Cftr chloride and Scnn1 sodium channels in the male submandibular correlated with greater NaCl reabsorption. In conclusion, adult salivary glands display sex‐ and gland‐specific differences in gene expression that reflect their unique functional properties.

Primary Cilium in the Human Thyrocyte: Changes in Frequency and Length in Relation to the Functional Pathology of the Thyroid Gland.

Primary cilia (PC) are conserved structures in the adult thyroid gland of different mammals. It was recently described that in humans, PC are usually present as a single copy per follicular cell emerging from the follicular cell apex into the follicular lumen. To understand the role developed by PC in thyroid hormonogenesis better, their changes in different human functional thyroid diseases (diffuse toxic hyperplasia/Graves' disease [GD] and nodular hyperplasia [NH]/nodular goiter), in comparison to normal thyroid tissue, were investigated using immunofluorescence, morphometry, and electron microscopy analyses. Significantly decreased ciliary frequencies were found in both NH (51.16 ± 11.69%) and GD (44.43 ± 23.70%) compared to normal thyroid tissue (76.09 ± 7.31%). Similarly, PC lengths were also significantly decreased in both NH (2.02 ± 0.35 μm) and GD (2.4 ± 0.48 μm) compared to normal glands (3.93 ± 0.90 μm). Moreover, in GD patients, hyperactive-follicle foci always showed diminished ciliary frequency and length compared to any other thyroid follicle pattern, independent of their thyroid status. Finally, in GD, the percentage of thyrocytes exhibiting PC in the "normal-appearance areas" was significantly lower in correspondence with the subsistence of signs of thyroid biosynthetic hyperactivity after long-term antithyroid drug treatment. The results suggest a direct relationship between ciliogenesis and both follicle activity and tissue heterogeneity in the functional pathology of the thyroid gland.

Vector Competence of Thrips Species to Transmit Soybean Vein Necrosis Virus.

Soybean vein necrosis virus (SVNV) is a newly discovered species of tospovirus infecting soybean plants that is transmitted by the primary vector, soybean thrips (Neohydatothrips variabilis), and two additional secondary vectors, tobacco thrips (Frankliniella fusca) and eastern flower thrips (F. tritici). This study was undertaken to elucidate the association between virus acquisition [6, 12, 24, and 48 h acquisition access period (AAP)] and transmission efficiency [12, 24, and 48 h inoculation access period (IAP)] in the primary vector, N. variabilis, and to examine the mechanisms of vector competence by analyzing the effect of AAP (6, 12, and 24 h) on virus infection in various tissues. In addition, we examined virus infection in tissues of the two secondary vectors. We found a significant effect of virus acquisition on transmission efficiency, transmission rate post 6 and 48 h AAP was significantly lower than 12 and 24 h AAP. Our analysis did not reveal a correlation between virus transmission rate and virus RNA in corresponding N. variabilis adults. On the contrary, N. variabilis adults harboring higher accumulation of the virus (>104) resulted in lower transmission rates. Analysis of SVNV infection in the tissues revealed the presence of the virus in the foregut, midgut (region 1, 2, and 3), tubular salivary glands and principal salivary glands (PSG) of adults of all three vector species, however, the frequency of infected tissues was highest in N. variabilis followed by F. fusca and F. tritici. The frequency of SVNV infection in individual tissues specifically the salivary glands was lowest after 6 h AAP compared to 12 and 24 h AAP. This finding is in agreement with the transmission assays, where significantly lower virus transmission rate was observed post 6 h AAP. In addition, N. variabilis adults with high PSG infection (12 and 24 h AAP) were likely to have high percentage of foregut and midgut region 2 infection. Overall, results from the transmission assays and immunolabeling experiments suggest that shorter AAP results in reduced virus infection in the various tissues especially PSG, which are important determinants of vector competence in SVNV-thrips interaction.

The tributyltin leads to obesogenic mammary gland abnormalities in adult female rats

Tributyltin chloride (TBT) is an obesogen associated with several complications. However, few investigations have evaluated TBT effects on adult mammary glands (MG). In this investigation, we assessed whether TBT’s obesogenic effects resulted in abnormal MG fat pad expansion and other irregularities. TBT was administered to female rats (100 ng/kg/day for 15 days via gavage), and their MG morphophysiological development was assessed. We further assessed the MG fat pad for PPARγ, ERα, and aromatase protein expression, as well as inflammation, oxidative stress (OS), apoptosis and fibrosis. Irregular MG morphological development such as lower TEB number, alveolar (AB), lobule and differentiation (DF) score were observed in TBT rats. TBT rats had abnormal MG fat accumulation as evidenced by increased numbers of hypertrophic adipocytes, triglyceride (TG) levels and PPARγ expression. A strong negative correlation between the MG obesogenic makers and TEB number, AB and DF score were observed in TBT rats. MG inflammation was observed in TBT rats. A positive correlation between the MG obesogenic markers and inflammation were observed. High ERα and aromatase expression were observed in MG of TBT rats. MG OS, apoptosis and fibrosis were present in the TBT rats. Additionally, a positive correlation between the MG obesogenic markers and OS were observed in TBT rats. Thus, these data suggest that obesogenic TBT effects led to MG irregularities in the adult female rats.