The development of the mammary gland is unique: the final stages of development occur postnatally at puberty under the influence of hormonal cues. Furthermore, during the life of the female, the ...mammary gland can undergo many rounds of expansion and proliferation. The mammary gland thus provides an excellent model for studying the 'stem/progenitor' cells that allow this repeated expansion and renewal. In this Review, we provide an overview of the different cell types that constitute the mammary gland, and discuss how these cell types arise and differentiate. As cellular differentiation cannot occur without proper signals, we also describe how the tissue microenvironment influences mammary gland development.
Cellular senescence is characterized by an irreversible cell cycle arrest and a pro‐inflammatory senescence‐associated secretory phenotype (SASP), which is a major contributor to aging and ...age‐related diseases. Clearance of senescent cells has been shown to improve brain function in mouse models of neurodegenerative diseases. However, it is still unknown whether senescent cell clearance alleviates cognitive dysfunction during the aging process. To investigate this, we first conducted single‐nuclei and single‐cell RNA‐seq in the hippocampus from young and aged mice. We observed an age‐dependent increase in p16Ink4a senescent cells, which was more pronounced in microglia and oligodendrocyte progenitor cells and characterized by a SASP. We then aged INK‐ATTAC mice, in which p16Ink4a‐positive senescent cells can be genetically eliminated upon treatment with the drug AP20187 and treated them either with AP20187 or with the senolytic cocktail Dasatinib and Quercetin. We observed that both strategies resulted in a decrease in p16Ink4a exclusively in the microglial population, resulting in reduced microglial activation and reduced expression of SASP factors. Importantly, both approaches significantly improved cognitive function in aged mice. Our data provide proof‐of‐concept for senolytic interventions' being a potential therapeutic avenue for alleviating age‐associated cognitive impairment.
Senescence is a major contributor to aging and age‐related diseases. However, it is still unknown whether senolytics impact on cognitive function during the aging process. We found that both pharmacogenetic clearance of p16Ink4a senescent cells or treatment with senolytic cocktail Dasatinib and Quercetin, reduced senescent microglia in the hippocampus and improved cognitive function in aged mice.
Accurate inundation maps for flooded wetlands and rivers are a critical resource for their management and conservation. In this paper, we automate a method (thresholding of the short-wave infrared ...band) for classifying peak inundation in the Okavango Delta, northern Botswana, using Landsat imagery and Google Earth Engine. Inundation classification in the Okavango Delta is complex owing to the spectral overlap between inundated areas covered with aquatic vegetation and dryland vegetation classes on satellite imagery, and classifications have predominately been implemented on broad spatial resolution imagery. We present the longest time series to date (1990–2019) of inundation maps for the peak flood season at a high spatial resolution (30 m) for the Okavango Delta. We validated the maps using image-based and in situ data accuracy assessments, with overall accuracy ranging from 91.5% to 98.1%. Use of Landsat imagery resulted in consistently lower (on average, 692 km2) estimates of inundation extent than previous studies that used Moderate Resolution Imaging Spectroradiometer (MODIS) and National Oceanic and Atmospheric Administration Advanced Very-High-Resolution Radiometer (NOAA AVHRR) imagery, likely owing to the increased number of mixed pixels that occur when using broad spatial resolution imagery, which can lead to overestimations of the size of inundated areas. We provide the inundation maps and Google Earth Engine code for public use. This classification method can likely be adapted for inundation mapping in other regions.
Accurately estimating hippopotamus (Hippopotamus amphibius) numbers is difficult due to their aggressive nature, amphibious lifestyle, and habit of diving and surfacing. Traditionally, hippos are ...counted using aerial surveys and land/boat surveys. We compared estimates of numbers of hippos in a lagoon in the Okavango Delta, counted from land to counts from video taken from a DJI Phantom 4TM drone, testing for effectiveness at three heights (40 m, 80 m, and 120 m) and four times of day (early morning, late morning, early afternoon, and late afternoon). In addition, we determined effectiveness for differentiating age classes (juvenile, subadult, and adult), based on visual assessment and measurements from drone images, at different times and heights. Estimates in the pool averaged 9.18 (± 0.25SE, range 1-14, n = 112 counts). Drone counts at 40 m produced the highest counts of hippos, 10.6% higher than land counts and drone counts at 80 m, and 17.6% higher than drone counts at 120 m. Fewer hippos were counted in the early morning, when the hippos were active and most likely submerged, compared to all other times of day, when they tended to rest in shallow water with their bodies exposed. We were able to assign age classes to similar numbers of hippos from land counts and counts at 40 m, although land counts were better at identifying juveniles and subadults. Early morning was the least effective time to age hippos given their active behaviour, increasingly problematic with increasing height. Use of a relatively low-cost drone provided a rigorous and repeatable method for estimating numbers and ages of hippos, other than in the early morning, compared to land counts, considered the most accurate method of counting hippos.
The treelike structures of many organs, including the mammary gland, are generated by branching morphogenesis, a reiterative process of branch initiation and invasion from a preexisting epithelium. ...Using a micropatterning approach to control the initial three-dimensional structure of mouse mammary epithelial tubules in culture, combined with an algorithm to quantify the extent of branching, we found that the geometry of tubules dictates the position of branches. We predicted numerically and confirm experimentally that branches initiate at sites with a local minimum in the concentration of autocrine inhibitory morphogens, such as transforming growth factor-β. These results reveal that tissue geometry can control organ morphogenesis by defining the local cellular microenvironment, a finding that has relevance to control of invasion and metastasis.
Wildlife rehabilitation is a critical part of animal welfare that contributes to species conservation. Despite the resources that go into rehabilitation, how animals fare after release from care is ...unknown. This is particularly true for cryptic arboreal species where specialist diets in care and low detectability in the wild present challenges for both care and post-release monitoring. We evaluated post-release outcomes for koalas and assessed if koalas were fed appropriately while in care. We monitored 36 koalas that had experienced one of three categories of medical intervention (none, minor, major) during rehabilitation. We examined the drivers of (i) koala survival and (ii) movements post-release, and (iii) evaluated variation between the species of browse fed in care versus browse selected by koalas in-situ. Overall, the post release survival rate of koalas was 58.5%, with only koalas that received medical intervention experiencing mortality. A critical threshold for mortality occurred at two weeks post-release and mortality was related to the measurable indicators of low body condition and poor climbing ability at time of release. In the month following their release, animals translocated furthest from their capture point moved the furthest. There was poor overlap between the tree species that koalas were fed in care and those they utilized post-release. We provide recommendations to address critical gaps in rehabilitation practices, as well as priorities for monitoring animals post-release to improve outcomes for arboreal folivores.
•THS exposure significantly increased pan-tumor incidence in CC mice.•THS exposure significantly reduced pan-tumor-free survival in CC mice.•Tumor susceptibility after THS exposure varied ...significantly across different CC strains.•Genetic background plays an important role in THS exposure-induced mouse tumorigenesis.
Increasing evidence has shown that thirdhand smoke (THS) exposure is likely to induce adverse health effects. An important knowledge gap remains in our understanding of THS exposure related to cancer risk in the human population. Population-based animal models are useful and powerful in investigating the interplay between host genetics and THS exposure on cancer risk. Here, we used the Collaborative Cross (CC) mouse population-based model system, which recapitulates the genetic and phenotypic diversity observed in the human population, to assess cancer risk after a short period of exposure, between 4 and 9 weeks of age. Eight CC strains (CC001, CC019, CC026, CC036, CC037, CC041, CC042 and CC051) were included in our study. We quantified pan-tumor incidence, tumor burden per mouse, organ tumor spectrum and tumor-free survival until 18 months of age. At the population level, we observed a significantly increased pan-tumor incidence and tumor burden per mouse in THS-treated mice as compared to the control (p = 3.04E-06). Lung and liver tissues exhibited the largest risk of undergoing tumorigenesis after THS exposure. Tumor-free survival was significantly reduced in THS-treated mice compared to control (p = 0.044). At the individual strain level, we observed a large variation in tumor incidence across the 8 CC strains. CC036 and CC041 exhibited a significant increase in pan-tumor incidence (p = 0.0084 and p = 0.000066, respectively) after THS exposure compared to control. We conclude that early-life THS exposure increases tumor development in CC mice and that host genetic background plays an important role in individual susceptibility to THS-induced tumorigenesis. Genetic background is an important factor that should be taken into account when determining human cancer risk of THS exposure.
Epithelial cell invasion through the extracellular matrix (ECM) is a crucial step in branching morphogenesis. The mechanisms by which the mammary epithelium integrates cues from the ECM with ...intracellular signaling in order to coordinate invasion through the stroma to make the mammary tree are poorly understood. Because the cell membrane-bound matrix metalloproteinase Mmp14 is known to play a key role in cancer cell invasion, we hypothesized that it could also be centrally involved in integrating signals for mammary epithelial cells (MECs) to navigate the collagen 1 (CL-1)-rich stroma of the mammary gland. Expression studies in nulliparous mice that carry a NLS-lacZ transgene downstream of the Mmp14 promoter revealed that Mmp14 is expressed in MECs at the tips of the branches. Using both mammary organoids and 3D organotypic cultures, we show that MMP activity is necessary for invasion through dense CL-1 (3 mg/ml) gels, but dispensable for MEC branching in sparse CL-1 (1 mg/ml) gels. Surprisingly, however, Mmp14 without its catalytic activity was still necessary for branching. Silencing Mmp14 prevented cell invasion through CL-1 and disrupted branching altogether; it also reduced integrin β1 (Itgb1) levels and attenuated MAPK signaling, disrupting Itgb1-dependent invasion/branching within CL-1 gels. FRET imaging revealed that Mmp14 associates directly with Itgb1. We identified a domain of Mmp14 that is required for modulating the levels of Itgb1, MEC signaling and the rate of invasion within CL-1. These results shed light on hitherto undescribed non-proteolytic activities of Mmp14 that are necessary for the Itgb1-dependent biochemical and mechanical signals that regulate branching in the mammary epithelium.
ABSTRACT
Breath-hold diving is a common practice as a part of military dive training. An association between prior lung injury and a propensity for lung barotrauma may have the potential to impact ...mission readiness for combat divers, Pararescue, Combat Controllers, Army Engineer divers, and various units in Naval Special Warfare and Special Operations. Barotrauma is a common complication of diving, typically occurring at depths greater than 30 m (98.4 ft). Individuals with abnormal lung anatomy or function may be at increased risk of barotrauma at shallower depths than those with healthy lungs, rendering these service members unfit for certain missions. We describe the case of a 25-year-old male, with a remote history of polytrauma and resultant pulmonary pleural adhesions, whose dive training was complicated by lung barotrauma at shallow depths. In missions or training utilizing breath-hold diving, the association with secondary alterations in lung or thoracic anatomy and function may limit which service members can safely participate.
Here, we describe a simple micromolding method to construct three-dimensional arrays of organotypic epithelial tissue structures that approximate in vivo histology. An elastomeric stamp containing an ...array of posts of defined geometry and spacing is used to mold microscale cavities into the surface of type I collagen gels. Epithelial cells are seeded into the cavities and covered with a second layer of collagen. The cells reorganize into hollow tissues corresponding to the geometry of the cavities. Patterned tissue arrays can be produced in 3-4 h and will undergo morphogenesis over the following 1-3 d. The protocol can easily be adapted to study a variety of tissues and aspects of normal and neoplastic development.
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