Transcriptomic Analysis of Mineralized Adipose-Derived Stem Cell Tissues for Calcific Valve Disease Modelling.

Calcific aortic valve disease (CAVD) is characterized by the fibrosis and mineralization of the aortic valve, which leads to aortic stenosis and heart failure. At the cellular level, this is due to the osteoblastic-like differentiation of valve interstitial cells (VICs), resulting in the calcification of the tissue. Unfortunately, human VICs are not readily available to study CAVD pathogenesis and the implicated mechanisms in vitro; however, adipose-derived stromal/stem cells (ASCs), carrying the patient's specific genomic features, have emerged as a promising cell source to model cardiovascular diseases due to their multipotent nature, availability, and patient-specific characteristics. In this study, we describe a comprehensive transcriptomic analysis of tissue-engineered, scaffold-free, ASC-embedded mineralized tissue sheets using bulk RNA sequencing. Bioinformatic and gene set enrichment analyses revealed the up-regulation of genes associated with the organization of the extracellular matrix (ECM), suggesting that the ECM could play a vital role in the enhanced mineralization observed in these tissue-engineered ASC-embedded sheets. Upon comparison with publicly available gene expression datasets from CAVD patients, striking similarities emerged regarding cardiovascular diseases and ECM functions, suggesting a potential link between ECM gene expression and CAVDs pathogenesis. A matrisome-related sub-analysis revealed the ECM microenvironment promotes the transcriptional activation of the master gene runt-related transcription factor 2 (RUNX2), which is essential in CAVD development. Tissue-engineered ASC-embedded sheets with enhanced mineralization could be a valuable tool for research and a promising avenue for the identification of more effective aortic valve replacement therapies.

SNAP-Ed Programming for College Students Experiencing Food Insecurity: A Qualitative Process-Focused Evaluation.

BACKGROUND: Growing recognition of food insecurity experienced by college students has led to efforts on college campuses to provide students with food assistance benefits and related nutrition education. A Supplemental Nutrition Assistance Program-Education (SNAP-Ed) program was developed for college students as one of these efforts. OBJECTIVE: To explore program content, recruitment, and implementation through experiences of program implementers in a novel SNAP-Ed intervention among college students. DESIGN: This qualitative study used focus groups and a case study approach to elicit program implementers' experiences delivering SNAP-Ed to college students. PARTICIPANTS/SETTING: Students, staff, and faculty (n = 26) implementing SNAP-Ed with college students experiencing food insecurity across eight campuses in the California State University system participated in 8 focus groups in 2021-2022. MAIN OUTCOME MEASURES: A process evaluation framework was used to evaluate content fit and orientation; recruitment, retention, and reach; and structure and capacity for implementation. ANALYSES PERFORMED: Focus groups were recorded, transcribed, and coded via thematic analysis using NVivo (QSR International, Burlington, MA). RESULTS: Five themes were identified: (1) need for this work to extend college-based basic needs services; (2) importance of aligning programming with college student context/needs; (3) common factors important for attracting/engaging the audience; (4) program barriers; and (5) training/team sharing for extending ideas. CONCLUSIONS: Study findings suggest tailoring SNAP-Ed programming to the needs of college students experiencing food insecurity, such as integrating student-relevant cooking skills, recipes, and topics of interest. Additional intervention and research efforts may lead to a new model for serving college students with SNAP-Ed.

Hyaluronidase: What is your fear?

BACKGROUND: The aesthetic treatment based on fillers with hyaluronic acid presents an increasing demand in the present day because it is considered a safe and minimally invasive procedure. In the management of adverse effects or more severe complications of hyaluronic acid-based fillers, hyaluronidase is the treatment of choice. AIMS: To demonstrate efficacy in reversibility and safety in the treatment of HA complications. PATIENTS/METHODS: It is a retrospective study article that reports the use of hyaluronidase in the main undesirable effects of fillers in 114 patients in a private dermatological clinic from 2015 to 2018. RESULTS: The target of the application was 51 cases of overcorrection (45%), 50 cases of Tyndall effect (44%), and 13 late nodules (11%). When we evaluated the areas where HYAL was injected, we found that the area with the most indication of the application of the product was the eyelid region (58 injections). CONCLUSION: This study concluded that HYAL is a safe and effective drug in the management of mild adverse events of HA applications with no severe side effects in our protocol of use.

Analysis of the Columellar-Labial Angle in Perioral Aging.

BACKGROUND: The young face presents a convex and rounded aspect; with aging, this changes to a more concave and flattened aspect, with the angles of the face undergoing changes through soft tissue repositioning and bone remodeling. The columellar-labial angle is one of the features that change. OBJECTIVES: The objective of the present study was to analyze the columellar-labial angle at different stages of adult life and assess its contribution to facial changes during aging. METHODS: We analyzed a database (December 2017-March 2018) of 722 female patients, aged 21 to 88 years, and obtained anthropometric measurements of the columellar-labial angle with a Canfield Vectra 3D image analysis system. Our database originates from a private clinic where all patients were registered and photographed. RESULTS: Our analysis showed a decrease in the columellar-labial angle, mainly observed from patients aged 60 years and older. This decrease can be explained by the absorption of body structures that occurs during aging. CONCLUSIONS: The columellar-labial angle decreases with age, and this finding may provide a better understanding of the transformation that occurs with aging. Obtaining a clearer view of the changes that occur in the face will improve treatments for facial rejuvenation, either conservative or surgical, and provide a basis for future studies and knowledge expansion.

Effect of the aging process on columella-labial, naso-mental and facial angles and how to apply it in clinical practice.

BACKGROUD: Aging is the result of the interplay of changes occurring in the facial skeleton, ligaments, muscles, adipose tissue, and skin (1), and these changes befall each mentioned structure at a different pace, start in each individual at a different age, and differ between ethnic background. (4) It changes directly the measurements of the face angles, making the face more concave and flattened aspect, while the young face presents a more convex and rounded aspect. Female skulls had an increase in all transverse facial widths and depth of the middle face with increasing age. Therefore, aging is consequence of progressive changes through intrinsic and extrinsic factors (5) and different methods can be used to assess and diagnose these changes. AIMS: The aim of the study is to evaluate facial angles and their aging changes. METHODS: A retrospective study was performed with a total of 1213 Caucasian female patients in the author's private clinic. The angles evaluate age-related changes were analyzed: facial, naso-mental, and columella-labial angle. Photograph analysis program Vectra 3D (Canfield) was used to perform the anthropometric assessment of the angles, and after data collection, the analysis was performed statistics. RESULTS AND CONCLUSION: We could conclude angles analyzed change with age, while two angles increase (naso-mental and facial angle), the other decreases (columella-labial angle). The study reinforces the principle of an objective clinical anthropometric assessment of facial angles as a relevant guide for a more appropriate treatment plan for the doctor to perform aesthetic procedures in order to restore a youthful face.

The Ovulatory Signal Precipitates LRH-1 Transcriptional Switching Mediated by Differential Chromatin Accessibility.

In the ovary, follicular growth and maturation are complicated processes that involve a series of morphological and physiological changes in oocytes and somatic cells leading to ovulation and luteinization, essential processes for fertility. Given the complexity of ovulation, characterization of genome-wide regulatory elements is essential to understand the mechanisms governing the expression of specific genes in the rapidly differentiating follicle. We therefore employed a systems biology approach to determine global transcriptional mechanisms during the early stages of the ovulatory process. We demonstrate that, following the hormonal signal that initiates ovulation, granulosa cells undergo major modification of distal regulatory elements, which coincides with cistrome reprogramming of the indispensable orphan nuclear receptor liver receptor homolog-1 (LRH-1). This cistromic reorganization correlates with the extensive changes in gene expression in granulosa cells leading to ovulation. Together, our study yields a highly detailed transcriptional map delineating ovarian cell differentiation during the initiation of ovulation.

Erosive Pustular Dermatosis of the Scalp: Why Do We Miss It?

Erosive pustular dermatosis of the scalp (EPDS) is an uncommon disease and primarily affects older men who have photo-damaged bald scalp, as was confirmed by our case series. EPDS is probably an overlooked disease, whose diagnosis is often missed because of a higher incidence of other cutaneous diseases affecting the same area and usually secondary to chronic actinic damage, such as actinic keratosis, basal cell carcinoma, and squamous cell carcinoma. For the first time, we report a case series of misdiagnosed EPDS with the aim of understanding why a diagnosis of EPDS was initially missed and try to give some tips to avoid future diagnostic delay.

Definition of the Mandibular Angle Using Botulinum Toxin Type A: A New Technique for the Improvement of Mandibular Contour and Definition.

While the injection of botulinum toxin type A (BoNTA) in the neck area, for face contouring through the blockade of the platysmal bundles, and in the mandibular body line has previously been explored, reports of its application for the demarcation of the mandibular angle are more limited. This article described a new technique of BoNTA injection in the mandibular angle for the improvement of contour and definition. At roughly 15 days after application in 30 patients, better visualization of the mandibular angle and its contours was evident and, consequently, smoothing of the appearance of sagging around the mandible with no adverse effects was noted. Further investigation of this technique is warranted.

Global Mapping of Open Chromatin Regulatory Elements by Formaldehyde-Assisted Isolation of Regulatory Elements Followed by Sequencing (FAIRE-seq).

Genetic information is organized in a complex structure composed of DNA and proteins together designated chromatin. Chromatin plays a dynamic role in transcriptional processes in that alteration of the interaction between its components results in the deregulation of cellular transcriptional program. Modification of epigenetic marks, variation in the precise positioning of nucleosomes, and consequent mobilization of nucleosomes regulate the access of various transcriptional factors to its underlying DNA template. Nucleosome-depleted regions, also designated open chromatin domains, are associated with active DNA regulatory elements, including promoters, enhancers, silencers, and insulators. Here, we describe the protocol of a rapid and simple technique entitled FAIRE (formaldehyde-assisted isolation of regulatory elements). Combined with high-throughput sequencing (FAIRE-seq), this procedure allows isolation of nucleosome-free regions and their mapping along the genome, thereby providing a global view of cell-specific regulatory elements.

The transcriptional co-repressor TLE3 suppresses basal signaling on a subset of estrogen receptor α target genes.

Chromatin constitutes a repressive barrier to the process of ligand-dependent transcriptional activity of nuclear receptors. Nucleosomes prevent the binding of estrogen receptor α (ERα) in absence of ligand and thus represent an important level of transcriptional regulation. Here, we show that in breast cancer MCF-7 cells, TLE3, a co-repressor of the Groucho/Grg/TLE family, interacts with FoxA1 and is detected at regulatory elements of ERα target genes in absence of estrogen. As a result, the chromatin is maintained in a basal state of acetylation, thus preventing ligand-independent activation of transcription. In absence of TLE3, the basal expression of ERα target genes induced by E2 is increased. At the TFF1 gene, the recruitment of TLE3 to the chromatin is FoxA1-dependent and prevents ERα and RNA polymerase II recruitment to TFF1 gene regulatory elements. Moreover, the interaction of TLE3 with HDAC2 results in the maintenance of acetylation at a basal level. We also provide evidence that TLE3 is recruited at several other regulatory elements of ERα target genes and is probably an important co-regulator of the E2 signaling pathway. In sum, our results describe a mechanism by which TLE3 affects ligand dependency in ERα-regulated gene expression via its binding restricting function and its role in gene regulation by histone acetylation.

LRH-1 governs vital transcriptional programs in endocrine-sensitive and -resistant breast cancer cells.

Tumor characteristics are decisive in the determination of treatment strategy for patients with breast cancer. Patients with estrogen receptor α (ERα)-positive breast cancer can benefit from long-term hormonal treatment. Nonetheless, the majority of patients will develop resistance to these therapies. Here, we investigated the role of the nuclear receptor liver receptor homolog-1 (LRH-1, NR5A2) in antiestrogen-sensitive and -resistant breast cancer cells. We identified genome-wide LRH-1-binding sites using ChIP-seq (chromatin immunoprecipitation sequencing), uncovering preferential binding to regions distal to transcriptional start sites. We further characterized these LRH-1-binding sites by integrating overlapping layers of specific chromatin marks, revealing that many LRH-1-binding sites are active and could be involved in long-range enhancer-promoter looping. Combined with transcriptome analysis of LRH-1-depleted cells, these results show that LRH-1 regulates specific subsets of genes involved in cell proliferation in antiestrogen-sensitive and antiestrogen-resistant breast cancer cells. Furthermore, the LRH-1 transcriptional program is highly associated with a signature of poor outcome and high-grade breast cancer tumors in vivo. Herein, we report the genome-wide location and molecular function of LRH-1 in breast cancer cells and reveal its therapeutic potential for the treatment of breast cancers, notably for tumors resistant to treatments currently used in therapies.

Endocrine resistance in breast cancer: from cellular signaling pathways to epigenetic mechanisms.

Although multiple cellular mechanisms have been proposed to explain endocrine resistance in breast cancer, the genomics events promoting the dysregulation of gene expression pattern are not clearly understood. Because chromatin plays a dynamic role in the estrogen receptor α (ERα) transcriptional program, we herein review signaling pathways implicated in endocrine resistance and try to merge them with recent epigenetic studies.

ERRs and cancers: effects on metabolism and on proliferation and migration capacities.

ERRs are orphan members of the nuclear receptor superfamily which, at least for ERRα and ERRγ display important roles in the control of various metabolic processes. On other hand, correlations have been found between the expression of ERRα and γ and diverse parameters of tumor progression in human cancers. Whereas it is tempting to speculate that ERR receptors act in tumors through the regulation of metabolism, recent data have suggested that they also may directly regulate tumor proliferation and progression independently of their effects on metabolism. The two aspects of tumoral functions of ERR receptors are the purpose of the present review.

H3K27 demethylation by JMJD3 at a poised enhancer of anti-apoptotic gene BCL2 determines ERα ligand dependency.

Chromatin represents a repressive barrier to the process of ligand-dependent transcriptional activity of nuclear receptors. Here, we show that H3K27 methylation imposes ligand-dependent regulation of the oestrogen receptor α (ERα)-dependent apoptotic response via Bcl-2 in breast cancer cells. The activation of BCL2 transcription is dependent on the simultaneous inactivation of the H3K27 methyltransferase, EZH2, and the demethylation of H3K27 at a poised enhancer by the ERα-dependent recruitment of JMJD3 in hormone-dependent breast cancer cells. We also provide evidence that this pathway is modified in cells resistant to anti-oestrogen (AE), which constitutively express BCL2. We show that the lack of H3K27 methylation at BCL2 regulatory elements due to the inactivation of EZH2 by the HER2 pathway leads to this constitutive activation of BCL2 in these AE-resistant cells. Our results describe a mechanism in which the epigenetic state of chromatin affects ligand dependency during ERα-regulated gene expression.

Modulating estrogen receptor-related receptor-alpha activity inhibits cell proliferation.

High expression of the estrogen receptor-related receptor (ERR)-alpha in human tumors is correlated to a poor prognosis, suggesting an involvement of the receptor in cell proliferation. In this study, we show that a synthetic compound (XCT790) that modulates the activity of ERRalpha reduces the proliferation of various cell lines and blocks the G(1)/S transition of the cell cycle in an ERRalpha-dependent manner. XCT790 induces, in a p53-independent manner, the expression of the cell cycle inhibitor p21(waf/cip)(1) at the protein, mRNA, and promoter level, leading to an accumulation of hypophosphorylated Rb. Finally, XCT790 reduces cell tumorigenicity in Nude mice.

The orphan receptor ERRalpha interferes with steroid signaling.

The estrogen receptor-related receptor alpha (ERRalpha) is an orphan member of the nuclear receptor superfamily that has been shown to interfere with the estrogen-signaling pathway. In this report, we demonstrate that ERRalpha also cross-talks with signaling driven by other steroid hormones. Treatment of human prostatic cells with a specific ERRalpha inverse agonist reduces the expression of several androgen-responsive genes, in a manner that does not involve perturbation of androgen receptor expression or activity. Furthermore, ERRalpha activates the expression of androgen response elements (ARE)-containing promoters, such as that of the prostate cancer marker PSA, in an ARE-dependent manner. In addition, promoters containing a steroid response element can be activated by all members of the ERR orphan receptor subfamily, and this, even in the presence of antisteroid compounds.

Endogenous cytotoxic T-cell response contributes to the long-term antiretroviral protection induced by a short period of antibody-based immunotherapy of neonatally infected mice.

Neutralizing monoclonal antibodies (MAbs) are increasingly being considered for blunting human viral infections. However, whether they can also exert indirect effects on endogenous antiviral immune responses has been essentially overlooked. We have recently shown that a short (several-day) period of immunotherapy with the neutralizing 667 MAb of mouse neonates shortly after infection with the lethal FrCas(E) retrovirus not only has an immediate effect on the viral load but also permits an endogenous antiviral immunity to emerge. Even though passive immunotherapy was administered during the particular period of immunocompetence acquisition, the endogenous response eventually arising was protective and persisted long (>1 year) after the MAb has disappeared. As very high levels of anti-FrCas(E) antibodies, predominantly of the immunoglobulin G2a (IgG2a) isotype and showing strong neutralization activity, were found in the sera of MAb-treated mice, it was necessary to address whether this humoral immunity was sufficient on its own to confer full protection against FrCas(E) or whether a cytotoxic T-lymphocyte (CTL) response was also necessary. Using a variety of in vivo assays in young and adult animals previously infected by FrCas(E) and treated by 667, we show here that transient 667 immunotherapy is associated with the emergence of a CTL response against virus-infected cells. This cytotoxic activity is indispensable for long-term antiviral protective immunity, as high neutralizing antibody titers, even enhanced in in vivo CD8(+) cell depletion experiments, cannot prevent the FrCas(E)-induced death of infected/treated mice. Our work may have important therapeutic consequences, as it indicates that a short period of MAb-based immunotherapy conducted at a stage where the immune system is still developing can be associated with the mounting of a functional Th1-type immune response characterized by both CTL and IgG2a-type humoral contributions, the cooperation of which is known to be essential for the containment of chronic infections by a variety of viruses.

Potentiation of ICI182,780 (Fulvestrant)-induced estrogen receptor-alpha degradation by the estrogen receptor-related receptor-alpha inverse agonist XCT790.

ICI182,780 (Fulvestrant) is a pure anti-estrogen used in adjuvant therapies of breast cancer. This compound not only inhibits the transcriptional activities of the estrogen receptor-alpha (ER alpha) but also induces its proteasome-dependent degradation. The latter activity is believed to be required for the antiproliferative effects of ICI182,780. Estrogen receptor-related receptor-alpha (ERR alpha) is an orphan member of the nuclear receptor superfamily that is expressed in a wide range of tissues including breast tumors, in which its high expression correlates with poor prognosis. Although not regulated by any natural ligand, ERR alpha can be deactivated by the synthetic molecule XCT790. Here we demonstrate that this compound also induces a proteasome degradation of ERR alpha. We also show that although it does not act directly on the steady-state level of ER alpha, XCT790 potentiates the ICI182,780-induced ER alpha degradation. We suggest that treatment with XCT790 could thus enhance the efficacy of ICI182,780 in ER alpha-dependent pathologies such as breast cancer.