Gap Closure Research Articles

G Protein-coupled Estrogen Receptor 1 (GPER1) Regulates Expression of SERPINE1/PAI-1 and Inhibits Tumorigenic Potential of Cervical Squamous Cell Carcinoma Cells In Vitro.

G protein-coupled estrogen receptor 1 (GPER1) appears to play a tumor-suppressive role in cervical squamous cell carcinoma (CSCC)GPER1 suppression leads to significantly increased expression of serpin family E member 1 (SERPINE1)/protein plasminogen activator inhibitor type 1 (PAI-1). The question arises, what role does SERPINE1/PAI-1 play in GPER1-dependent tumorigenic potential of CSCC. SiHa and C33A CSCC cells were treated with GPER1 agonist G1 or antagonist G36. SERPINE1/PAI-1 expression was suppressed by RNAi and success was confirmed by RT-qPCR. Protein expression of PAI-1 was quantified by Western blot. Viability was analyzed using resazurin assay, while migration was investigated using gap closure. Colony and tumor sphere formation were used to test clonogenicity. After G1 treatment, viability of SiHa and C33A cells remained unchanged. Cell migration was dose-dependently reduced. SiHa and C33A cells formed significantly fewer and smaller colonies as well as spheroids. Furthermore, treatment with G1 led to decreased expression of SERPINE1/PAI-1, while blockade of GPER1 with G36 resulted in significantly increased SERPINE1/PAI-1 expression. After suppression of SERPINE1/PAI-1 in SiHa cells using RNAi, cell viability remained unaffected; however, significantly smaller colonies were formed, and fewer and smaller spheroids were developed. Cell migration remained unaffected. Activation of GPER1 reduces clonogenicity and migration of CSCC cells and suppresses expression of SERPINE1/PAI-1. Suppression of SERPINE1/PAI-1 in CSCC cells reduces tumorigenic potential. GPER1 may be a suitable target for suppression of SERPINE1/PAI-1 in CSCC. However, SERPINE1/PAI-1 does not appear to be the decisive factor for GPER1-regulated cell migration.

Vibron Softening of Solid Hydrogen under Nanoconfinement.

The vibron behavior of hydrogen bears significant importance for understanding the phases of solid hydrogen under high pressure. In this work, we reveal an unusual high-pressure behavior of hydrogen confined within nanopores through a combination of experimental measurements and theoretical calculations. The nanoconfined hydrogen molecules retain an hcp lattice up to 170 GPa, yet significant deviations from the vibrational characteristics of bulk hydrogen are observed in the primary vibrons of both Raman and infrared spectra. This lowered vibron peak is linked to the disorder of the hydrogen molecules with longer bonds and enhanced intermolecular interactions at the interface. Further investigation reveals that this nanoscale confinement leads to a considerable decrease in the band gap of solid hydrogen, potentially facilitating band gap closure at considerably lower pressures. Our findings provide crucial insights into the behavior of solid hydrogen under spatial nanoconfinement, paving the way for novel explorations into hydrogen metallization.

Gender Gap Closure and Reversal

Gender Gap Closure and Reversal

Outcomes of Endoscopic versus Microscopic Ossicular Chain Reconstruction-A Systematic Review and Meta-analysis.

The efficacy of the transcanal endoscopic approach in tympanoplasty, cholesteatoma removal, and stapedotomy is detailed. This study reviews evidence on the feasibility of endoscopic ossiculoplasty as a suitable alternative to traditional microscopic ossiculoplasty. PubMed, Embase, and Cochrane Library were searched from inception to 19 June 2024 for articles comparing endoscopic and microscopic ossicular chain reconstruction. This systematic review was conducted in accordance with PRISMA guidelines. The primary outcomes assessed include air bone gap closure, pre-operative and post-operative air bone gap, pure tone audiometry, and word recognition scores. The secondary outcomes include operation time, surgical success, and surgical complications. These outcomes between microscopic and endoscopic techniques were pooled and compared as mean differences using a random-effects model. The Risk of Bias in Non-Randomized Studies of Interventions (ROBINS-I) and Cochrane Risk-of-Bias Tool for Randomized Trials (RoB2) scale were adopted to assess for study bias in observational studies and randomized controlled trials, respectively. This study included five studies comprising 504 patients. While endoscopic ossiculoplasty had shorter operating durations (95% CI -42.97 to -1.57), it conferred comparable air bone gap closure (95% CI -0.34 to 3.76), post-operative pure tone audiogram (95% CI 10.97-18.77), and surgical success. Additional qualitative strengths identified in endoscopic ear surgery include improvements in visualization, decreased need for supplemental incisions, excellent ergonomics, and reduced post-operative pain. Overall, endoscopic ossiculoplasty may confer comparable audiological outcomes to microscopic ossiculoplasty and may serve as a reasonable alternative surgical approach. Laryngoscope, 2025.

The Effectiveness and Safety of Tibial-Sided Osteotomy for Fibula Untethering in Lateral Close-Wedge High Tibial Osteotomy: A Novel Technique with Video Illustration.

Background and Objectives: Despite its advantages, lateral close-wedge high tibial osteotomy (LCWHTO) requires proximal tibiofibular joint detachment (PTFJD) or fibular shaft osteotomy for gap closing. These fibula untethering procedures are technically demanding and not free from the risk of neurovascular injuries. Our novel fibula untethering technique, tibial-sided osteotomy (TSO) near the proximal tibiofibular joint (PTFJ), aims to reduce technical demands and the risk of injury to the peroneal nerve and popliteal neurovascular structures. The purposes of this study were to introduce the TSO technique and compare the complexity and safety of TSO with those of radiographic virtual PTFJD, which is defined based on radiographic landmarks representing the traditional PTFJD technique. Materials and Methods: Between March and December 2023, 13 patients who underwent LCWHTO with TSO for fibula untethering were enrolled. All patients underwent MRI preoperatively and CT scanning postoperatively. The location of the TSO site on the postoperative CT scans was matched to preoperative MRI to measure the shortest distance to the peroneal nerve and popliteal artery. These values were compared with estimates of the distance between the PTFJ and neurovascular structures in the radiographic virtual PTFJD group. The protective effect of the popliteus muscle was evaluated by extending the osteotomy direction toward the posterior compartment of the knee. Results: The TSO procedure was straightforward and reproducible without producing incomplete gap closure during LCWHTO. On axial images, the distances between the surgical plane and the peroneal nerve or popliteal artery were significantly longer in the TSO group than in the radiographic virtual PTFJD group (both p = 0.001). On coronal and axial MRI, the popliteus muscle covered the posterior osteotomy plane in all patients undergoing TSO but did not cover the PTFJD plane in the radiographic virtual PTFJD group. Conclusions: Our novel TSO technique for fibula untethering during LCWHTO is reproducible and reduces the risk of neurovascular injury by placing the separation site more medially than in the PTFJD procedure.

Mesenchymal stem cell extracellular vesicle vascularization bioactivity and production yield are responsive to cell culture substrate stiffness

AbstractMesenchymal stem cell‐derived extracellular vesicles (MSC EVs) are an attractive therapeutic option for regenerative medicine applications due to their inherently pro‐angiogenic and anti‐inflammatory properties. However, reproducible and cost‐effective production of highly potent therapeutic MSC EVs is challenging, limiting their translational potential. Here, we investigated whether the well‐characterized responsiveness of MSCs to their mechanical environment—specifically, substrate stiffness—could be exploited to generate EVs with increased therapeutic bioactivity without the need for biochemical priming or genetic manipulation. Using polydimethylsiloxane and bone marrow‐derived MSCs (BM‐MSCs), we show that decreasing the stiffness of MSC substrates to as low as 3 kPa significantly improves the pro‐angiogenic bioactivity of EVs as measured by tube formation and gap closure assays. We also demonstrate that lower substrate stiffness improves EV production and overall yield, important for clinical translation. Furthermore, we establish the mechanoresponsiveness of induced pluripotent stem cell‐derived MSC (iMSC) EVs and their comparability to BM‐MSC EVs, again using tube formation and gap closure assays. With this data, we confirm iMSCs' feasibility as an alternative, renewable cell source for EV production with reduced donor variability. Overall, these results suggest that utilizing substrate stiffness is a promising, simple, and a potentially scalable approach that does not require exogenous cargo or extraneous reagents to generate highly potent pro‐angiogenic MSC EVs.

Enhancing food security while reducing environmental impacts: Life cycle assessment of cultivation-irrigation systems and yield gap closure in paddy fields

Enhancing food security while reducing environmental impacts: Life cycle assessment of cultivation-irrigation systems and yield gap closure in paddy fields

Public Valuations of Managing Compromised Molars: A Discrete Choice Experiment.

There is limited evidence to support optimal patient-centered management for compromised first permanent molars (cFPM) in children. Based on an online discrete choice experiment (DCE), this study elicits UK adult general population preferences and calculates willingness to pay (WTP) for pathways to manage cFPM. The DCE was designed with information from semistructured interviews and literature reviews, as well as focus groups with an expert panel of dentists, citizens, and policy makers. A statistically optimal D-efficient design generated 18 choice tasks, split across 2 blocks. Each respondent answered one block of 9 tasks to reduce survey fatigue. Choice tasks varied across 5 attributes: type of treatment, provider of care, who makes the management decision, number of future visits avoided, and cost. An opt-out was included (no treatment). Conditional logit models (fixed effects) were used for data analysis, and marginal WTP for each attribute level was calculated. An overall 430 respondents completed the DCE. Respondents valued children receiving care as compared with not. Restoring a cFPM was valued equally to spontaneous or orthodontic gap closure. In contrast, having a partial gap, prosthetic replacement with a bridge, or a full unit gap was valued less than restoration or full gap closure. General dentists were preferred to dentists with enhanced skills, but there was no evidence of a preference for general dentists over specialists in pediatric dentistry. Respondents preferred to be wholly or partly involved in the decision-making process as opposed to the dentist making the decision alone. Respondents preferred less costly treatments and the avoidance of future dental work. Dental care service providers must consider service user preferences for health and nonhealth outcomes in any service redesign. Furthermore, the results provide marginal WTP estimates that can be used to value dental care services.

Clinical and radiographical evaluation of two agenesis alveolar ridges of upper lateral incisors. A case report with 5 years follow up

Besides the functional issues, lateral incisor agenesis represents a significant aesthetic drawback (1). The decision about the treatment plan is complex due to the long-term success, which is not easy to achieve. It has to be discussed with the orthodontists to choose the closure, using permanent canines to replace the missing teeth, or maintaining the spaces, for example, with Maryland bridges or implants. The main advantage of orthodontic gap closure is that this approach preserves the natural architecture of the complex and soft tissues. However, canines and premolar sizes and shapes should be adjusted to mimic the replaced teeth, eventually resorting to odontoplasty or veneers (13). This article aims to show clinical protocols and procedures for the Split Crest without any graft. The biggest problem in agenesis cases is the thickness of the bone, which is bucco-palatal and associated with the need for proper functional and esthetic rehabilitation. In the case that we reported, because of the lack of thickness, we performed a minimally invasive split crest, which allowed us to correct the implant insertion with immediate loading by a temporary crown, just for aesthetic reasons.

Fingolimod Inhibits C6 Rat Glioma Proliferation and Migration, Induces Sub-G1 Cell Cycle Arrest, Mitochondrial and Extrinsic Apoptosis InVitro and Reduces Tumour Growth InVivo.

Glioblastoma multiforme (GBM), the most prevalent brain tumour, is universally fatal. GBM cells exhibit cell cycle disruption and treatment resistance, remarking an urgent need for newer treatments. Fingolimod, a sphingosine-1-phosphate receptor modulator, has been reported to have anti-cancer effects. This study investigated the therapeutic potentials of fingolimod in rat C6 cells and pursued the involved mechanism(s). Cell survival, proliferation, migration, and morphology of fingolimod-treated C6 cells were evaluated using MTT, soft-agar colony formation, wound-healing, and Giemsa staining assays. Apoptosis was investigated through acridine orange/ethidium bromide (AO/EB) and annexin V staining, and flow cytometry analysed the cell cycle. Quantitative reverse transcription PCR and western blotting were used to evaluate gene and protein expressions. An intracranial C6 rat model validated the anti-tumour effect of fingolimod. Fingolimod significantly reduced the survival and colonies of the C6 cells and delayed their gap closure. Cell shrinkage coupled with AO/EB and PI staining of the fingolimod-treated cells indicated apoptosis, subsequently confirmed by measuring the expression levels of the candidate genes involved in apoptosis and cell cycle, such as Bax/Bcl2, P53, Cytochrome C and Caspases 9/3, Fas, Fadd, Tnfrsf1a, Cdkn1a, and Ccnd1, at RNA and protein levels, indicating both extrinsic and mitochondrial apoptosis and cell cycle arrest at sub-G1 phase in fingolimod-treated cells. Furthermore, treating rats bearing intracranial C6 tumours with fingolimod led to significant suppression of intracranial tumour growth. Based on our findings, cell cycle arrest and apoptosis contribute to fingolimod antitumor effects.

Highly Biocompatible Lamellar Liquid Crystals Based on Hempseed or Flaxseed Oil with Incorporated Betamethasone Dipropionate: A Bioinspired Multi-Target Dermal Drug Delivery System for Atopic Dermatitis Treatment.

Atopic dermatitis (AD) is the most common chronic inflammatory skin disease that severely impairs patient's life quality and represents significant therapeutic challenge due to its pathophysiology arising from skin barrier dysfunction. Topical corticosteroids, the mainstay treatment for mild to moderate AD, are usually formulated into conventional dosage forms that are impeded by low drug permeation, resulting in high doses with consequent adverse effects, and also lack properties that would strengthen the skin barrier. Herein, we aimed to develop biomimetic lamellar lyotropic liquid crystals (LLCs), offering a novel alternative to conventional AD treatment. In screening studies, pseudoternary phase diagrams alongside polarized light microscopy (PLM) and viscosity measurements were utilized. Next, the selected LCCs underwent comprehensive characterization via PLM, small-angle X-ray scattering, differential scanning calorimetry, and rheological analysis. Lastly, their performance was evaluated and compared with the commercially available reference medicine in chemical stability study, in vitro permeation testing, in vitro safety assessment using cell proliferation assay, inverted light microscopy, and Raman mapping of keratinocytes, besides gap closure assay performed by live-cell imaging. Formulation (L/T)Ho30, containing the highest amount of lecithin/Tween 80 mixture (21%) and hempseed oil (28%), demonstrated lamellar microstructure with high skin hydration potential and favourable rheological features for skin administration. Moreover, in comparison with the reference medicine, it stood out by providing suitable chemical BD (betamethasone dipropionate) stability, improved 3-fold BD permeation, and excellent biocompatibility with over 85% cell proliferation at all tested concentrations, ensuring keratinocytes' integrity, as well as promoting skin healing with gap closure observed after 36hours. Unique multi-target drug delivery strategy depicted in newly developed bioinspired lamellar LCCs structurally resembling stratum corneum intercellular lipids, with incorporated BD drug, and composed of multifunctional components that synergistically strengthen skin barrier, was presented here and shows a promising approach for improved AD treatment.

Coupled Hydro-Gas-Mechanical 3D Modeling of LASGIT Experiment

Gas transport simulation in bentonite for radioactive waste disposal poses challenges for numerical models due to its complex microstructure. Understanding the processes involved is a prerequisite for assessing gas flow's impact on repository layouts. The DECOVALEX23 (D-2023) Task B (Large Scale Gas Injection Test: LASGIT) project aims to advance numerical techniques for predicting gas flow in repository systems through gas injection tests on compacted bentonite at the British Geological Survey (BGS). This study develops a comprehensive coupled hydro-gas-mechanical 3D numerical model to simulate the test, considering heterogeneous initial permeability and embedded fractures. Addressing bentonite swelling, three gap closure scenarios for the canister-bentonite blocks gap interface were considered. The model reproduces observed test behaviors, capturing preferential gas flow paths. Sensitivity analysis explores variations in volume factor sensitivity, calibration, hydraulic conductivity of interfaces, heterogeneity, permeability, and model parameters, contributing to a deeper understanding of the phenomenon's complexity. The proposed hydraulic modeling, enriched by considerations of gap closure states, predicts measured evolution of gas injection trends. suggesting reliability and potential applicability for similar conditions and facilitating a comprehensive analysis of its impact on gas testing processes. Additionally, the embedded fracture models underscore the critical role of fracture behavior and dilatancy in determining the system's hydro-mechanical response, with significant sensitivity to these factors influencing stress and pore pressure evolution. Hydro-mechanical models demonstrate that modeling approaches involving embedded fractures and dilatancy significantly influence gas pathways and entry gas pressure. System volume plays pivotal role in the analysis, while sensitivity analysis of contact transmissivity reveals potential influences on preferential gas pathway formation. Hydraulic and hydro-mechanical modeling methods show promise for further numerical investigations, indicating potential for yielding meaningful insights in future studies.

Determination of Optimum Irrigation Water Requirement of Wheat at Sayo District, Kellem Wollega, Oromia

Ethiopia needs 70-80 million quintals of wheat to feed 110 million people, but imports are a significant deficit. The government has implemented food and wheat security measures, including yield gap closure, area expansions, and irrigation. Wheat is a strategic commodity for Ethiopia's food security, agroindustry, import substitution, and job creation. An experiment was conducted at Dambi Dollo University campus in Sayo District, Kellem Wollega Zone, to determine wheat irrigation water demand. The study evaluated five irrigation depths in wheat cultivation, using soil parameters, meteorological data, and crop characteristics. The results showed that 90% of the net water requirement of wheat is optimal for normal physiological activities, including evapotranspiration and metabolic activities. The saved water, 10% of the total 191.9 mm, can be used to irrigate more command areas and prevent abandonment. The water productivity value of irrigated wheat is within the range of previous findings, and the average wheat yield is improved under irrigation conditions even in Ethiopia. The efficiency parameters (GY, WUE, and WP) generally perform significantly. The study recommends extension services to demonstrate this finding on farmers' fields for further evaluation and popularization, and Zonal/District Agricultural Offices to use this 172.71 mm to sustainably boost irrigated wheat production.

Impact of pharmacy involvement on care gap closure in Managed Medicaid patients.

A population health pharmacy team was developed to improve health and increase reimbursement for patients with value-based care plans. The purpose of this study was to evaluate the impact of pharmacy involvement on care gap closure in Managed Medicaid patients. This was a single-center, retrospective cohort study of Managed Medicaid patients seen at outpatient facilities. Patients had a pharmacy risk score (PRS) of 6 or greater and had not achieved the Medicaid quality measures for both glycated hemoglobin (HbA1c) and blood pressure (BP). The intervention group included patients reviewed by pharmacy, compared to a control group of patients not reviewed by pharmacy. The primary outcome was closure of at least one care gap by the end of 2022. Secondary outcomes were the number of each type of gap closed, the frequency at which recommendations were made by pharmacists, and the frequency at which pharmacist recommendations were implemented by providers. Data were collected for 80 patients for the period from January through October 2022. The primary outcome occurred in 37 (74%) patients in the intervention group and 15 (50%) patients in the control group (odds ratio, 2.85; P = 0.032). The HbA1c gap was closed in 30 (60%) patients in the intervention group and 8 (27%) patients in the control group. The BP gap was closed in 24 (48%) patients in the intervention group and 11 (37%) patients in the control group. The frequency with which recommendations were made by a pharmacist was associated with gap closure (P = 0.012). No significant difference was found based on the frequency at which recommendations were implemented by providers (P = 0.4). Pharmacy intervention was associated with an almost 3-fold-higher likelihood of closing at least one care gap in Medicaid patients. HbA1c gap closure was achieved more frequently than BP gap closure due to pharmacy involvement. The frequency with which recommendations were made by pharmacy was associated with increased gap closure regardless of the frequency with which these recommendations were implemented by providers.

Hubbard Gap Closure-Induced Dual-Redox Li-Storage Mechanism as the Origin of Anomalously High Capacity and Fast Ion Diffusivity in MOFs-Like Polyoxometalates.

MOFs-like polyoxometalate (POMs) electrodes, harvesting combined advantages of interlocking porosity and multi-electron transfer reaction, have already emerged as promising candidates for lithium-ion batteries (LIBs), yet the origins of the underlying redox mechanism in such materials remain a matter of uncertainty. Of critical challenges are the anomalously high storage capacities beyond their theoretical values and the fast ion diffusivity that cannot be satisfactorily comprehended in the theory of crystal lattice. Herein, for the first time we decode t2g electron occupation-regulated dual-redox Li-storage mechanism as the true origin of extra capacity in POMs electrodes. The lattice and electronic transition of active centers and reaction intermediates were systematically decoupled through density functional theory (DFT) and a suite of structural spectroscopic investigations, such as X-ray absorption near-edge spectroscopy (XANES), soft X-ray absorption spectroscopy (sXAS) and 7Li solid-state nuclear magnetic resonance (NMR). Enhanced V-t2g orbital occupation by Li coordination significantly triggers the Hubbard gap closure and reversible Li deposition/dissolution at surface region. Conjugated V-O-Li configuration at interlayers endow Li+ ion pathways along pore walls as the dominant contribution to the low migration barrier and fast diffusivity. As a result, remarkable cycle stability (~100 % capacity retention after 2000 cycles at 1 A g-1), extremely high specific capacity (1200 mAh g-1 at 100 mA g-1) and excellent rate performance (404 mAh g-1 at 8 A g-1) were achieved, providing new understandings on the underlying mechanism of POMs electrodes and pivotal guidance for dual-storage materials.

Endoscopic Fat Graft Myringoplasty Augmented With Hyaluronic Acid for Managing Large-Sized Eardrum Perforations; A Prospective Comparative Randomised Study.

We aimed to assess the outcomes of fat graft myringoplasty augmented with hyaluronic acid in closing large-sized eardrum perforations compared to the traditional underlay cartilage-perichondrium composite myringoplasty (CPCM). It was a prospective randomised comparative study. It was held in tertiary referral institutions between May 2020 and April 2022. We included 100 patients with a large-sized eardrum perforation (50%-75% of the eardrum surface area). Using the endoscopic transcanal approach, 50 patients were managed by fat graft myringoplasty augmented with hyaluronic acid, while CPCM managed the other 50 patients. We evaluated the closure rates 1, 6 months and 1 year after surgery. Also, we assessed the audiological performance of the patients with a successful closure before and 1 year after the operation. Fat graft myringoplasty operation was statistically shorter than the CPCM. The closure rate 1 year after surgery was 92% in the first group and 86% in the second group, without a statistically significant difference between both groups. Successful air-bone gap closure to less than 10 dB occurred in 93.5% of group A and 81.4% of group B without a statistically significant difference. The mean postoperative air-bone gap was 5.3 ± 3.95 dB in the first group and 7.95 ± 5.17 dB in the second group, with a statistically significant difference. Fat graft myringoplasty augmented with hyaluronic acid was a reliable, safe, simple, and effective manoeuvre to close large-sized eardrum perforations compared to the conventional CPCM.

Simulation of homogenization behavior of compacted bentonite containing technological voids using modified penalty-based contact model

Low-density zones generated during the bentonite blocks/voids homogenization process in the repository may serve as potentially preferential paths for radionuclide leakage. More importantly, void closure during homogenization process involves complex contact problems, where the stiffness at the contact interface typically undergoes significant fluctuations. In this work, with contact interface stiffness addressed through a step function approach, a modified penal.ty-based contact model was proposed to simulate the contact behavior involved at the gap closure stage of the bentonite/gap assemblage homogenization process. Then, unsaturated infiltration swelling tests on bentonite block (1.7 Mg/m3)/gap (width: 2 mm) assemblages were performed, and the variation of dry density at different hydrated times (0, 24, 72, 168, and 720 h) in specific areas were measured. Based on the results, time-dependent swelling pressure profiles of the assemblage were acquired, while the homogenization process was evaluated. Results reveal that after approximately 40 h of hydration, the gap is completely closed, and the radial stress condition of the compacted bentonite transits progressively from the initial free swelling into a constant volume expansion state. The swelling pressure correspondingly develops quickly to a peak value at 1.8 MPa once the hydration starts, then decreases to a valley value of 1.4 MPa at the complete gap closure, and subsequently begins to increase to the final stable value of 1.8 MPa. Further examination reveals that as hydration advances, dry density of the assemblage converges to the expected final dry density with a maximum residual inhomogeneity of about 2 %. Finally, validations demonstrate that the proposed model can accurately reproduce deformations of the assemblage during the free swelling stage, and the swelling pressure profiles. A comparative analysis was made with the previous approach of identifying gaps as highly deformable materials, revealing that the proposed model overcomes the traditional limitations associated with the separation or penetration behavior occurring between the compacted bentonite and contact boundaries during the gap closure.

Evaluation of Storage Parameters of Platelet-Rich Plasma in Functional Assay of Epithelial Gap Repair

Platelet-rich plasma (PRP), produced from autologous blood and containing growth factors, has shown promise in treating various conditions, including diabetic wounds. However, as expected, patient compliance is low in the context of repeated treatments as required for hard-to-heal wounds. This study investigates the efficacy of PRP stored under refrigeration for up to 2 weeks in promoting in-vitro human epithelial cell gap closure. PRP was prepared from six healthy individuals and immediately split into sterile test tubes for storage at 4°C; the impact of fresh PRP on epithelial gap closure was compared to paired PRP samples stored for one and two weeks. According to gap closure analysis, 5 out of 6 individuals depicted accelerated epithelial gap closure compared to control conditions. There was no significant difference between fresh and refrigerated samples. An overall benefit was observed for using 1-week old samples over 2-week-old samples. Interestingly, according to a 6-hr epithelial gap analysis, significantly accelerated cell migration was obtained by 1-week-old refrigerated samples compared to their corresponding fresh PRP samples. The current study provides data in support of increasing the accessibility of PRP treatment for patients with hard-to-heal wounds. Patients can potentially self-treat between wound clinic appointments by using refrigerated sterile aliquots of their fresh PRP. Notably, by the end of two weeks, refrigerated PRP will not act as fresh PRP. More research is needed to fully investigate the effect of refrigeration on PRP sample efficacy, including larger cohorts, clinical wounds, and an evaluation of the molecular fingerprint of refrigerated PRP samples.

Effects of BET Inhibition on lung micro-environmental changes in obesity-related pulmonary Arterial Hypertension (PAH)

Abstract Background Obesity is emerging as a pivotal risk factor for pulmonary arterial hypertension (PAH) but the underlying molecular mechanisms are poorly explored. The lung microenvironment, namely the cell-cell communication among fibroblasts, immune and endothelial cells, may significantly impact on vascular structure and function thus promoting microvascular disease in this setting. Obesity-induced chromatin modifications lead to maladaptive transcriptional programs altering cell function in different organs. Apabetalone (APA), a selective inhibitor of bromodomain and extra-terminal containing protein family (BET) proteins, prevents bromodomain-containing protein 4 (BRD4) interactions with chromatin thus modulating gene expression. Purpose To determine whether a new epi-drug, namely APA, affects the lung microenvironment in obesity-related PAH. Methods Mice were fed a normal diet (ND, control) or high-fat-diet (HFD) and L-NAME for 15 weeks to induce obesity-related PAH (obPAH). Echocardiography was performed. Pathways regulating obPAH were identified by single-cell nuclei RNA sequencing in lung specimens. Primary endothelial cells (ECs) and Fibroblasts (mFs) were freshly isolated from obPAH lungs and treated with APA for 48 hours. Cells and lung samples were used for molecular biology and histology. Cellular migration was investigated by in vitro gap closure. Results Obese mice displayed PAH as shown by increased pulmonary artery (PA) pressure, PA resistance, right ventricular wall thickness and reduced PA acceleration time. Of note, chronic APA treatment prevented PAH-related features in obese mice. To determine the cell types affected by APA, we performed scRNAseq. We found that fibroblast and endothelial cells had the highest enrichment of genes whose expression significantly correlated with echo-determined PAH features. ECs and mFs from obese mice displayed an altered phenotype whereas treatment with APA rescued obesity-driven ECs and mFs dysfunction. In obPAH ECs, we found increased expression of NF-kB p65-related inflammatory genes (IL-1β, IL-6, TNF-alpha) as well as enhanced p53 signaling. Moreover, obPAH-ECs expressed high levels of hypoxia-inducible factor (HIF1-α) and NADPH-oxidase-NOX4 with subsequent increase in ROS generation. Furthermore, APA treatment protected against cellular inflammation, senescence and oxidative stress. To evaluate the effect of APA and the paracrine response of fibroblasts, we performed a scratch assay on control mFs exposed to TGF-β or conditioned media collected from obPAH-ECs with and without APA. In both experimental settings, APA affected gap closure, suggesting that the treatment influences fibroblast migration through both a direct effect and paracrine mechanisms. Conclusions APA is able to reset the lung micro-envirnment in obesity thus reducing inflammation and senescence. BET inhibitors could represent potential therapeutic approaches in this setting.

Suppressing Expression of SERPINE1/PAI1 Through Activation of GPER1 Reduces Progression of Vulvar Carcinoma.

The serine proteinase inhibitor 1 (SERPINE1) gene codes for the plasminogen activator inhibitor 1 (PAI1) protein and is thought to play a tumor supportive role in various cancers. In this work we aimed to uncover the role PAI1 plays in the proliferation, migration, and invasion of vulvar cancer (VC), and define the protein's function as an oncogene or tumor suppressor. Through treatment with an agonist (G1) and antagonist (G36) of G-coupled estrogen receptor 1 (GPER1), an upstream regulator of SERPINE1 expression, and a forward transfection knockdown protocol, the expression of SERPINE1/PAI1 in VC cells was altered. The effects these altered SERPINE1/PAI1 levels had on tumor cell functions were then examined. Proliferation was analyzed using the resazurin assay, while migration was studied via the gap closure assay. Through colony- and tumor sphere- formation assays clonogenicity was tested, and western blots showed protein expression. In A431 VC cells, when the levels of PAI1 were reduced via knockdown or treatment with G1, migration, proliferation, and colony growth was reduced. Treatment with G36 increased expression of PAI1 and increased migration and colony size in CAL39 cells. Based on the findings in this study, suppressing PAI1 expression in VC cells appears to reduce their progression and tumorigenic potential. Therefore, PAI1 could possibly function as an oncogene in VC. GPER1 appears to be a suitable target for suppressing PAI1 in VC.