bluuebunny/crossref_metadata_2025_split huggingface dataset:
Susceptibility of Hepatic ProgenitoInhibition by acetazolamide of reactivated carbonic anhydrase activity in a receptor protein tyrosine phosphatase γ (RPTPγ) mutantr Cells to Lipotoxicity as Contributor for Impaired Liver Regeneration and Disease Advancement In Pediatric Nonalcoholic Fatty Liver Disease (PNAFLD)
Authors: Viswanathan Preeti , Maisuradze luka , Jaber Fadi‐luc , Gupta Sanjeev 
Published: May 2022

BackgroundIn PNAFLD mechanisms underlying early onset and rapid disease progression have been unclear. Active liver growth in preadolescent children imparted by facultative progenitor cells would increase genome damage following oxidative stress and lipotoxicity, leading to DDR and further hepatic injury events. To reveal this we considered hTERT‐FH‐B hepatocytes expressing progenitor properties will be ideal for modeling growth‐related lipotoxicity and DDR of PNAFLD.Methods/ResultsThe FH‐B cells were derived from late‐gestation liver with doubling time of 24h, including 20‐30% in S phase, which was similar to pH3 histone +ve proliferating liver cell fractions in 5‐8‐yr old healthy children. Moreover, FH‐B cells express hepatobiliary markers (G6P, Glycogen, GGT, DPPIV) along with progenitor markers (SOX9, FOXA2, Cx43, EpCAM) and P450 profiles indicating hepatic immaturity. To induce lipotoxicity in cultured cells 50 μM palmitic acid (PA) was added with and without oxidized collagen matrix or 2,5‐10 ng/ml TNFa. This dose‐dependently caused oxidative stress (dihydrorhodamine dye for ROS) and lower mitochondrial membrane potential (MMP, JC‐1 dye) in 1‐4h and onset of steatosis (Oil red O or Bodipy staining) and decreased cell viability (MTT assay) in 18‐24h. The contribution of ATM signaling during lipotoxicity was apparent from pNBS1 accumulation, which marks DNA strand break sites. Moreover, lipotoxicity and DNA damage greatly increased in presence of ATM‐specific kinase inhibitor, KU‐60019. Additionally, ATM insufficiency exacerbated cell growth‐arrest through downstream p53‐dependent p21 activation, which increased polyploidy indicated by Hoechst dye‐stained nuclear DNA content analysis, and inhibited cell number increases in growth assays over few days. To test therapeutic feasibility, we studied obeticholic acid (OCA) and found this drug decreased ATM insufficiency‐related exacerbation of lipotoxicity. The improvements included steatosis, ROS, MMP. DDR, cell viability, as well as less growth‐arrest or polyploid accumulations and continued cycling.ConclusionsThe successful modeling of PNAFLD in hTERT‐FH‐B cells establishes the critical susceptibility of actively growing facultative progenitor hepatocytes to mitochondrial dysfunction and impaired DDR related to ATM insufficiency. The potential of OBC in improving lipotoxic outcomes will be relevant for PNAFLD. These pathophysiology‐based models offer opportunities to advance therapy in PNAFLD.
PFKFB3 Mediated Changes in ROS/NO Balance Contribute to Endothelial Dysfunction in Obesity
Authors: Batori Robert K. , Bordan Zsuzsanna , Padgett Caleb , Atawia Reem , de Chantemele Eric Belin , Stepp David , Fulton David J. 
Published: May 2022

Obesity is a strong risk factor for cardiovascular disease, in large part due to an altered metabolic state. One of the earliest consequences of cardiovascular dysfunction in obesity is the loss of endothelial function and impaired nitric oxide (NO) signaling. In blood vessels, NO is synthesized by endothelial nitric oxide synthase (eNOS). The biological actions of NO can be compromised by rapid inactivation by reactive oxygen species (ROS) such as superoxide that is mainly generated by NADPH oxidases (NOXs). However, the signals coordinating the increased superoxide production leading to loss of NO with altered metabolism remain unknown.We have found that the expression of 6‐phosphofructo‐2‐kinase/fructose‐2, 6‐biphosphatase 3 (PFKFB3), which is a key glycolytic regulatory enzyme, is significantly increased in endothelial cells of obese animals. Overexpression of PFKFB3 within the endothelium of blood vessels is accompanied by the loss of vascular relaxation and in decreased NO production as a result of increased eNOS phosphorylation on the inhibitory T495 residue. PFKFB3 overexpression also blunted Akt‐S473 phosphorylation, reducing stimulus‐dependent phosphorylation and activation of eNOS at S1177. While PFKFB3 exerted negative effects on NO signaling, it increased the activity and mRNA levels of NOX1, a major contributor to endothelial dysfunction. Further, we have also found that pharmacological inhibition of NOX1 attenuates the decreased vasodilator responses in PFKFB3 overexpressing aortic rings.These results demonstrate a novel functional relationship between endothelial metabolism, ROS and NO production that may contribute to endothelial dysfunction in obesity.
ATP Attenuates α1‐Adrenergic and Peptidergic Vasocontraction in Isolated Porcine Cerebral Arteries
Authors: Luchkanych Adam M. , Morse Cameron , Tomczak Corey R. , Olver T. D. 
Published: May 2022

IntroductionCompeting influences regulate blood flow control in the brain. In the skeletal muscle vasculature, purinergic ATP signaling attenuates α1‐adrenergic and peptidergic‐induced vascular smooth muscle contraction (i.e., phenomenon referred to as functional sympatholysis); however, whether this occurs in the cerebrovasculature remains unknown. The purpose of this experiment was to examine the effect of ATP on vascular responses to α1‐adrenergic‐ and peptidergic‐receptor activation in cerebral arteries. We hypothesized that ATP would attenuate the α1‐adrenergic‐ and peptidergic‐mediated vasocontraction in isolated pial arteries.MethodsFemale pigs (n=5) were euthanized and their brains harvested. Thereafter, 1A branches of the middle cerebral artery were dissected for wire‐myography. Dose‐response curves for the α1‐adrenergic agonist phenylephrine (PE;1e‐10‐1e‐4M) and the peptidergic agonist neuropeptide Y (NPY;1e‐12‐1e‐6M) were performed in the absence or presence of ATP (1e‐6M). Paired one‐tailed t‐tests were used to compare the overall magnitude of contraction (area under the curve; AUC) and physiological maximal responses between conditions.ResultsData are mean±SD. The AUC of PE‐mediated cerebral vasocontraction was attenuated by ATP (untreated=67±25 vs. ATP pre‐treatment=33±23 AU; p=0.03). However, the mean reduction in the maximal response to PE in arteries pre‐treated with ATP was not significant (untreated=43±15 vs. ATP pre‐treatment=27±11%; p=0.10). The AUC (untreated=62±31 vs. ATP pre‐treatment=37±23 AU) as well as maximal NPY‐mediated cerebral vasocontraction (untreated=65±20 vs. ATP pre‐treatment=49±21%) was attenuated by ATP (all p≤0.05).ConclusionThese data indicate ATP attenuates PE‐ and NPY‐mediated vasocontraction in isolated pial arteries. Thus, similar to the skeletal muscle vasculature, purinergic signaling attenuates vasoreactivity to α1‐adrenergic and peptidergic receptor activation in the cerebrovasculature. Functional sympatholysis may assist in coupling cerebral blood flow to brain metabolism under basal conditions as well as in the setting of heightened sympatho‐excitation.
Specificity of Binding and Antitumor Efficacy of a Covalent Microtubule Stabilizer
Authors: Risinger April L. , Yee Samantha S. 
Published: May 2022

Microtubule stabilizers, including the taxanes paclitaxel and docetaxel, are a mainstay in the treatment of solid tumors, including breast and ovarian cancers. Although these agents are often effective as first‐line therapy, they are susceptible to common drug resistance mechanisms, including the upregulation of drug efflux pumps. Therefore, there is a critical need to develop novel classes of microtubule targeted agents that retain activity in taxane‐resistant settings. The C22,23‐epoxy taccalonolides are a class of plant‐derived microtubule stabilizers that covalently and irreversibly bind microtubules, allowing them to retain efficacy in a variety of taxane‐resistant cancer models. We employed cellular‐based binding studies to fully map the taccalonolide pharmacophore and confirm the target specificity of this covalent compound. The identification of β‐tubulin residues that are critical for taccalonolide binding in a cellular context has provided the opportunity to generate functional taccalonolide mimetics through a synthetically tractable approach.Herein, we demonstrate that taccalonolide AF is less susceptible to p‐glycoprotein‐mediated drug efflux than paclitaxel in two separate ovarian cancer models and that it has superior cellular persistence after drug washout, likely due to irreversible target engagement. While taccalonolide AF demonstrated potent and persistent in vivo antitumor efficacy against the taxane‐resistant NCI/ADR‐RES ovarian cancer model as a flank xenograft when administered systemically, it was associated with a narrow therapeutic window. Additional studies showing that intratumoral injection of the taccalonolides into flank tumors provided long‐term antitumor efficacy months after the final dose prompted a study of their utility in the targeted delivery to a disseminated orthotopic ovarian cancer model representing localized metastasis. Indeed, we found that taccalonolide‐treated animals showed a significant decrease in micrometastasis of taxane‐resistant ovarian cancer cells to the spleen, as detected by quantitative RT‐PCR, without any evidence of systemic toxicity. We propose that the irreversible mechanism of microtubule stabilization elicited by the taccalonolides has the unique potential for intraperitoneal treatment of locally disseminated taxane‐resistant disease, which represents a significant unmet clinical need in the treatment of ovarian cancer patients.
The role of cytoskeleton regulators in embryonic facial morphogenesis
Authors: Ibrahim Isra , Richman Joy M. 
Published: May 2022

Small GTPases are downstream of the non‐canonical WNT JNK‐PCP pathway. The most well studied members of this group are RhoA, Rac1 and CDC42. These small GTPases regulate cell shape, orientation, and migration through regulating the actomyosin cytoskeleton. In addition, components of this pathway have been linked to non‐syndromic cleft lip with or without cleft palate. The objective of this study is to test whether small GTPase signaling is required and sufficient for facial morphogenesis at stages where lip fusion is taking place.MethodsThe faces of E4.5 chicken embryos were dissected to include the two facial processes involved in lip fusion, the frontonasal and maxillary processes (Fig. 1). After a dose‐response experiment to find the optimal concentrations, the organ cultures were treated with either an antagonist of RhoA signaling (ROCKi 20µM) or an antagonist of Rac1 (Rac1i 100µM) or an agonist (RhoA/ Rac1/CDC42 activator 0.5µg/ml). Organ cultures were photographed and measured at 0 and 24hrs. Percentage growth differences were analyzed using Prism Graphpad software (v. 9.2).ResultsIn the control cultures, frontonasal mass width decreased by 23% and increased in cranio‐caudal height by 17% over 24‐hour period. The nasal slits did not change. The maxillary prominences increased in height and width by 12% and 37% respectively over 24‐hour period similar to growth in vivo. The ROCKi treated cultures prevented the narrowing of the frontonasal mass and increased the length of the nasal slits. ROCKi treatment had no effect on the height of the frontonasal mass nor maxillary prominences. In contrast, the RAC1i had no effect on facial growth. The agonist significantly reduced frontonasal mass width compared to ROCKi. In addition, the agonist significantly decreased growth of the maxillary prominences.ConclusionsThe changes in facial morphogenesis may be explained by active cytoskeletal remodeling, particularly in the RhoA pathway. We excluded a requirement for RAC1 in facial morphogenesis in our system. Our data also suggest that different regions of the face have different requirements for specific small GTPase signaling. The frontonasal mass is dependent on RhoA signaling but not the maxillary prominences. Over activation of all GTPases shows that correct levels are necessary for normal morphogenesis. Overall, we have revealed a complex interplay between small GTPAses in midfacial morphogenesis.
Morphological Studies of the Left Ventricle in the Creatine Deficiency Mice Model
Authors: Shults Nataliia V. , de Souza Aline M. , Baydyuk Maryna , Sandberg Kathryn , Huang Jeffrey K. 
Published: May 2022

IntroductionHeart failure is the major cause of death characterized by changes in myocardial energetics, including a reduced level of creatine. In organs with high energy demand such as the heart, creatine is taken up by cardiomyocytes via creatine transporter with subsequent transformation into phosphocreatine, which controls by creatine kinase. This system functions as shuttles high‐energy phosphates from mitochondria to contractile elements such as the myofibril, which utilize 70 % of total produced energy by mitochondria. For the past decades, many studies tried to understand whether a reduced level of creatine affects the myocardial function leading to the failing heart. However, most of the results from these studies have been contradictory. The present study describes the detailed morphologic changes of cardiomyocytes of the left ventricle (LV) of mice with whole‐body creatine deficiency due to knockout of the biosynthetic enzyme (guanidinoacetate N‐methyltransferase, GAMT).Methods & ResultsThe present study used GAMT‐/‐ mice model with the unique metabolic fingerprint of zero creatine similar to those in human GAMT deficiency. The histological and transmission electron microscopy (TEM) studies of LV myocardium in GAMT‐/‐ mice showed cardiac hypertrophy and increased collagen accumulation in the myocardial interstitium compared to the wild‐type (WT) mice. The TEM examination showed irreversible destructive ultrastructure changes of cardiomyocytes in conjunction with insufficient adaptive protein synthesis accompanied by focal and diffuse lysis of myofilaments in cardiac myocytes. The mitochondrial changes were characterized by cristae homogenization and fragmentation in association with cluster formation. The volume of density of mitochondria (VvMt), myofibrils (VvMf), and their ratios (VvMt/VvMf), along with the coefficient of energy efficiency of mitochondria (CEEM) have been examined in GAMT‐/‐ mice in comparison to WT. We established that VvMt, VvMf, VvMt/VvMf, and CEEM were all significantly reduced. Further, destructive changes in the nucleus and nuclei, focal degradation of sarcoplasm, increases autophagy, are all evidence of inadequate adaptive protein synthesis in cardiac myocytes.ConclusionsThese studies of GAMT‐/‐ mice demonstrated that creatine deficiency promotes decreased VvMt/VvMf and CEEM in LV myocytes, which may contribute to the development progression of heart failure and in conjunction with insufficient adaptive protein synthesis contribute to irreversible ultrastructure restructuring of cardiomyocytes with loss contractile myocardial function.
Novel Role of Dual Oxidase 2 as a Mediator of Podocyte Injury in the Diabetic Environment
Authors: Castro Aracely , Ford Bridget M. 
Published: May 2022

Glomerular injury is a prominent pathological feature of diabetic nephropathy (DN). In glomerular epithelial cells, or podocytes, hyperglycemia alters slit diaphragm proteins and causes foot process effacement, apoptosis and cell detachment. Oxidative stress has emerged as an important pathogenic mechanism in the development of glomerular injury in DN. However, the mechanisms by which these factors exert their action remain poorly understood.We provide evidence that the NADPH oxidase of the Nox family, Dual oxidase 2 (Duox2), is present in cultured glomerular cells, including podocytes. Exposure of cultured podocytes to high concentrations of glucose (HG) elicited a rapid upregulation of Duox2 protein expression. Inhibition of Duox2 with specific siRNA prevented the HG‐induced increase in intracellular reactive oxygen species (ROS) generation and hydrogen peroxide (H2O2) production in cultured podocytes.In additional experiments, we established a functional link between Duox2‐derived ROS generation and podocyte injury in response to HG. Impairment of Duox2 function nearly abolished HG‐mediated DNA fragmentation, apoptosis and decrease in/reorganization of slit diaphragm protein expression in podocytes, indicating that Duox2 is required for the deleterious effects of glucose in podocytes.Our study demonstrates, for the first time, that Duox2 is responsible for increased ROS generation and subsequent alteration of podocyte function in response to HG. Our work serves as proof of concept to demonstrate the utility of targeting Duox2 as a future therapeutic intervention to reduce diabetes‐mediated glomerular lesions.
Individualized Out-of-Pocket Price Estimators for “Shoppable” Surgical Procedures: A Nationwide Cross-Sectional Study of US Hospitals
Authors: Berlin Nicholas L. , Chopra Zoey , Bryant Arrice , Agius Josh , Singh Simone R. , Chhabra Karan R. , Schulz Paul , West Brady T. , Ryan Andrew M. , Kullgren Jeffrey T. 
Published: May 2022

Objective:
To estimate the nationwide prevalence of individualized out-of-pocket (OOP) price estimators at US hospitals, characterize patterns of inclusion of 14 specified “shoppable” surgical procedures, and determine hospital-level characteristics associated with estimators that include surgical procedures.

Background:
Price transparency for shoppable surgical services is a key requirement of several recent federal policies, yet the extent to which hospitals provide online OOP price estimators remains unknown.

Methods:
We reviewed a stratified random sample of 485 U.S. hospitals for the presence of a tool to allow patients to estimate individualized OOP expenses for healthcare services. We compared characteristics of hospitals that did and did not offer online price estimators and performed multivariable modeling to identify facility-level predictors of hospitals offering price estimator with and without surgical procedures.

Results:
Nearly two-thirds (66.0%) of hospitals in the final sample (95% confidence interval 61.6%–70.1%) offered an online tool for estimating OOP healthcare expenses. Approximately 58.5% of hospitals included at least one shoppable surgical procedure while around 6.6% of hospitals included all 14 surgical procedures. The most common price reported was laparoscopic cholecystectomy (55.1%), and the least common was recurrent cataract removal (20.0%). Inclusion of surgical procedures varied by total annual surgical volume and health system membership. Only 26.9% of estimators explicitly included professional fees.

Conclusions:
Our findings highlight an ongoing progress in price transparency, as well as key areas for improvement in future policies to help patients make more financially informed decisions about their surgical care.
A Rare Anatomical Finding of Undescribed Accessory Palmaris Longus in the Distal Forearm: A Case Report
Authors: Allahham Salim , Alyazji Zaki T. N. , Aljassem Ghanem , Sada Ruba , Wani Iqbal Rasool 
Published: May 2022

Summary:
Upper limb muscle anomalies and their clinical implications have been described frequently in the literature reviews. In this article, we are presenting a case of aberrant forearm muscle that had not been described before, and could be considered as a palmaris longus muscle variation. A 24-year-old man presented to the emergency department, Hamad General Hospital, Doha, Qatar, with right forearm laceration with multiple cut structures for which he was admitted for exploration and repair. Intraoperatively, flexor digitorum superficialis of the third, fourth, and fifth digits, flexor carpi radialis, and palmaris longus were injured‚ and all of them were repaired. We noticed an aberrant muscle—which was also injured—that originated from the distal third of the radius on its medial aspect to insert into the palmar fascia; pulling this muscle’s tendon resulted in tightening of palmar fascia‚ same as the palmaris longus. Along with the importance of deep knowledge of typical human anatomy, hand surgeons must be aware that an aberration from normal anatomy might be anticipated, to provide the best care to our patients.
EFFECTS OF ECTOPIC INNER FOVEAL LAYERS ON FOVEAL CONFIGURATION AND VISUAL FUNCTION AFTER IDIOPATHIC EPIRETINAL MEMBRANE SURGERY
Authors: Yang Xiaohan , Wang Zengyi , Yu Yanping , Mo Bin , Zhang Ke , Wu XiJin , Liu Lingzi , Wang Xinbo , Qi Biying , Liu Wu 
Published: May 2022

Purpose:
To investigate the effects of ectopic inner foveal layers (EIFLs) on foveal configuration recovery and visual acuity after idiopathic epiretinal membrane removal and analyze the relationship of foveal configuration recovery between 1-month and long-term postoperatively.

Methods:
This retrospective study included 216 consecutive eyes with idiopathic epiretinal membrane that underwent vitrectomy surgery. A comprehensive ophthalmic examination was performed for all patients before and 1, 4, 10, and 24 months after surgery. We observed the postoperative anatomical and functional recovery of eyes with and without EIFLs and analyzed the association between short-term and long-term anatomical recovery.

Results:
The presence of EIFL was associated with severe preoperative visual damage (P < 0.001) and was considered as a negative factor for postoperative foveal depression recovery (P < 0.001). No significant difference was found in postoperative best-corrected visual acuity between the eyes with and without EIFLs (P = 0.442). For eyes with Stage II epiretinal membranes, 17.6% developed EIFLs postoperatively, which did not affect the final best-corrected visual acuity. Overall, 24.5% of epiretinal membranes restored the foveal configurations at 1 month postoperatively, and 14.7% continued recovering during the follow-up. Foveal structure recovery in eyes with EIFLs was associated with earlier stage, fewer microcystic macular edema, complete interdigitation zone layer, and a thinner central foveal thickness (all P < 0.05) at 1 month postoperatively.

Conclusion:
The EIFL is associated with severe preoperative visual damage and is a negative factor for postoperative anatomical recovery, but it is a minor factor for postoperative visual acuity. We also found an association of foveal configuration recovery between 1 month and long-term after surgery.
IMPACT OF COVID-19 PANDEMIC LOCKDOWNS ON VISUAL ACUITY OF PATIENTS WITH NEOVASCULAR AMD
Authors: Shor Reut , Segal Ori , Mimouni Michael , Greenbaum Eran , Zur Dinah , Trivizki Omer , Schwartz Shulamit , Loewenstein Anat , Goldstein Michaella , Rabina Gilad 
Published: May 2022

Purpose:Evaluating the impact of delayed care, secondary to coronavirus disease 2019 (COVID-19) pandemic lockdowns, on visual acuity in previously treated neovascular age-related macular degeneration (nAMD) patients.Methods:This was a multicenter, retrospective, study of patients with nAMD previously treated with anti-VEGF injections who were followed up during 2019 (pre–COVID-19) and compared with patients with nAMD during 2020 (COVID-19).Results:A total of 1,192 patients with nAMD with a mean age of 81.5 years met the inclusion criteria. Of these, 850 patients were assessed in 2019 (pre–COVID-19) and 630 patients were assessed in 2020 (COVID-19). Three hundred eight patients were assessed through both 2019 and 2020 and thus were included in both cohorts. There was no significant difference between 2020 and 2019 in baseline and change in best-corrected visual acuity (BCVA;P= 0.342 andP= 0.911, respectively). The mean number of anti-VEGF injections was significantly lower (5.55 vs. 6.13,P< 0.01), with constant lower ratio of injections per patient in the COVID-19 period. Baseline BCVA (0.859,P< 0.01), number of injections (−0.006,P= 0.01), and age (0.003,P< 0.01) were predictors of final BCVA.Conclusion:In patients with nAMD, delayed care secondary to COVID-19 pandemic lockdowns has no statistically significant impact on BCVA. Best-corrected visual acuity, older age, and lower number of yearly anti-VEGF injections are predictors for decrease BCVA.