Validation of Autism Spectrum Disorder Diagnoses in Large Healthcare Systems with Electronic Medical Records

Journal of Autism and Developmental Disorders - To identify factors associated with valid Autism Spectrum Disorder (ASD) diagnoses from electronic sources in large healthcare systems. We examined...     

Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium

The endometrium is the inner lining of the uterus. Following specific cyclic hormonal stimulation, endometrial stromal fibroblasts (stroma) and vascular endothelial cells exhibit morphological and biochemical changes to support embryo implantation and regulate vascular function, respectively. Herein, we integrated a resin-based porous membrane in a dual chamber microfluidic device in polydimethylsiloxane that allows long term in vitro co-culture of human endometrial stromal and endothelial cells. This transparent, 2-μm porous membrane separates the two chambers, allows for the diffusion of small molecules and enables high resolution bright field and fluorescent imaging. Within our primary human co-culture model of stromal and endothelial cells, we simulated the temporal hormone changes occurring during an idealized 28-day menstrual cycle. We observed the successful differentiation of stroma into functional decidual cells, determined by morphology as well as biochemically as measured by increased production of prolactin. By controlling the microfluidic properties of the device, we additionally found that shear stress forces promoted cytoskeleton alignment and tight junction formation in the endothelial layer. Finally, we demonstrated that the endometrial perivascular stroma model was sustainable for up to 4 weeks, remained sensitive to steroids and is suitable for quantitative biochemical analysis. Future utilization of this device will allow the direct evaluation of paracrine and endocrine crosstalk between these two cell types as well as studies of immunological events associated with normal vs. disease-related endometrial microenvironments.     

Adrenal Steroids Modulate the Immune Response during Brucella abortus Infection by a Mechanism That Depends on the Regulation of Cytokine Production

Human brucellosis is a protean disease with a diversity of clinical signs and symptoms resulting from infection with Brucella species. Recent reports suggest a cross-regulation between adrenal steroids (cortisol and dehydroepiandrosterone [DHEA]) and the immune system. Monocytes and macrophages are the main replication niche for Brucella. Therefore, we investigated the role of adrenal hormones on the modulation of the immune response mediated by macrophages in B. abortus infection. Cortisol treatment during B. abortus infection significantly inhibits cytokine, chemokine, and MMP-9 secretion. In contrast, DHEA treatment had no effect. However, DHEA treatment increases the expression of costimulatory molecules (CD40, CD86), the adhesion molecule CD54, and major histocompatibility complex class I (MHC-I) and MHC-II expression on the surface of B. abortus-infected monocytes. It is known that B. abortus infection inhibits MHC-I and MHC-II expression induced by gamma interferon (IFN-γ) treatment. DHEA reverses B. abortus downmodulation of the MHC-I and -II expression induced by IFN-γ. Taken together, our data indicate that DHEA immune intervention may positively affect monocyte activity during B. abortus infection.     

Deconstructing cartilage shape and size into contributions from embryogenesis,metamorphosis, and tadpole and frog growth

Understanding skeletal diversification involves knowing not only how skeletal rudiments are shaped embryonically, but also how skeletal shape changes throughout life. The pharyngeal arch (PA) skeleton of metamorphosing amphibians persists largely as cartilage and undergoes two phases of development (embryogenesis and metamorphosis) and two phases of growth (larval and post‐metamorphic). Though embryogenesis and metamorphosis produce species‐specific features of PA cartilage shape, the extents to which shape and size change during growth and metamorphosis remain unaddressed. This study uses allometric equations and thin‐plate spline, relative warp and elliptic Fourier analyses to describe shape and size trajectories for the ventral PA cartilages of the frog Xenopus laevis in tadpole and frog growth and metamorphosis. Cartilage sizes scale negatively with body size in both growth phases and cartilage shapes scale isometrically or close to it. This implies that most species‐specific aspects of cartilage shape arise in embryogenesis and metamorphosis. Contributions from growth are limited to minor changes in lower jaw (LJ) curvature that produce relative gape narrowing and widening in tadpoles and frogs, respectively, and most cartilages becoming relatively thinner. Metamorphosis involves previously unreported decreases in cartilage size as well as changes in cartilage shape. The LJ becomes slightly longer, narrower and more curved, and the adult ceratohyal emerges from deep within the resorbing tadpole ceratohyal. This contrast in shape and size changes suggests a fundamental difference in the underlying cellular pathways. The observation that variation in PA cartilage shape decreases with tadpole growth supports the hypothesis that isometric growth is required for the metamorphic remodeling of PA cartilages. It also supports the existence of shape‐regulating mechanisms that are specific to PA cartilages and that resist local adaptation and phenotypic plasticity.     

Myoepithelial Cell Differentiation Markers in Ductal Carcinoma in Situ Progression

We describe a preclinical model that investigates progression of early-stage ductal carcinoma in situ (DCIS) and report that compromised myoepithelial cell differentiation occurs before transition to invasive disease. Human breast cancer MCF10DCIS.com cells were delivered into the mouse mammary teat by intraductal injection in the absence of surgical manipulations and accompanying wound-healing confounders. DCIS-like lesions developed throughout the mammary ducts with full representation of human DCIS histologic patterns. Tumor cells were incorporated into the normal mammary epithelium, developed ductal intraepithelial neoplasia and DCIS, and progressed to invasive carcinoma, suggesting the model provides a rigorous approach to study early stages of breast cancer progression. Mammary glands were evaluated for myoepithelium integrity with immunohistochemical assays. Progressive loss of the myoepithelial cell differentiation markers p63, calponin, and α-smooth muscle actin was observed in the mouse myoepithelium surrounding DCIS-involved ducts. p63 loss was an early indicator, calponin loss intermediate, and α-smooth muscle actin a later indicator of compromised myoepithelium. Loss of myoepithelial calponin was specifically associated with gain of the basal marker p63 in adjacent tumor cells. In single time point biopsies obtained from 16 women diagnosed with pure DCIS, a similar loss in myoepithelial cell markers was observed. These results suggest that further research is warranted into the role of myoepithelial cell p63 and calponin expression on DCIS progression to invasive disease.Clinical evidence is compelling for histologic progression of breast cancer through atypical hyperplasia, ductal carcinoma in situ (DCIS), invasive ductal carcinoma, and metastatic stages.¹ Such histopathologic progression studies and mutational profiling of epithelial cancers^{2, 3} suggest that acquisition of invasive potential is a relatively late event. However, genomic data analyses have revealed that most tumor cell gene expression changes occur at the transition from normal to DCIS, with few additional changes in expression occurring at the transition from DCIS to overt invasive disease.^{4, 5} These observations implicate key roles for nonepithelial cells in progression to invasive disease.^{6, 7} The lack of relevant model systems has hindered our understanding of the DCIS to invasive transition.The clinical definition of invasive breast cancer is spread of malignant tumor cells from the confines of the mammary duct into the adjacent tissue stroma. In the normal mammary gland, epithelial ductal and alveolar structures are surrounded by a contractile myoepithelial cell layer that facilitates milk expulsion during lactation.⁸ The mammary myoepithelial cells are also required for normal mammary gland development, because they influence epithelial cell polarity, ductal branching, and milk production.⁸ A hallmark of progression from DCIS to invasive cancer is physical breach of the myoepithelial cell layer and underlying basement membrane. For tumor progression, studies suggest that myoepithelial cells play an active role in tumor suppression by secreting protease inhibitors, down-regulating matrix metalloproteinases,^{9, 10} and producing tumor suppressive proteins such as maspin, p63, Wilms tumor 1, and laminin 1.^{11, 12, 13} These data support the hypothesis that the tumor suppressive function of myoepithelium is lost with DCIS progression, resulting in the transition from preinvasive to invasive cancer.^{14, 15, 16} Further studies report that tumor cells adjacent to focally disrupted myoepithelium can display distinct phenotypes, including estrogen receptor negativity, genetic instabilities, increased expression of invasion-related genes, and aberrant E-cadherin expression.^{17, 18} Overall, these data support an active role for the myoepithelium in suppressing DCIS progression and implicate loss of this function as critical for the transition to invasive disease.Invasive potential of human mammary epithelial tumor cell lines is evaluated primarily by injecting cells into the mammary fat pads of immune compromised mice. Although the mammary fat pad is the correct anatomic organ for breast cancer, mammary fat pad models bypass the requirement for tumor cells to exit from the location of their initiation, that is, the mammary ducts. In transgenic models, early-stage disease is intraductal, and these models display tumor progression from ductal intraepithelial neoplasia (DIN) to invasive stages. However, in transgenic models, most epithelial cells contain the active oncogene; thus, these models do not replicate cellular transformation as a relatively rare event. Here, we used an intraductal approach in the absence of surgery,¹⁹ because this approach offers a key advantage in that cells are directly placed into the mammary ductal system, which is the site of early-stage disease. Importantly, this approach permits modeling of disease progression in the background of a normal mammary epithelium. Further, our nonsurgical approach permits co-evolution of tumor progression with myoepithelial cell changes with minimal wound healing or proinflammatory induction. With this intraductal model, we observed progressive loss of the myoepithelial cell differentiation markers p63, calponin, and α-smooth muscle actin (α-SMA) before tumor cell breach of the myoepithelium. Further, myoepithelial cell loss of calponin strongly associates with gain of p63 expression in adjacent epithelial tumor cells, a marker of basal epithelium. These studies identify compromised myoepithelial cell function before transition to invasive disease and suggest that disrupted myoepithelial expression of calponin may predict DCIS-involved ducts at risk of progression to invasive disease.