bluuebunny/crossref_metadata_2025_split huggingface dataset:
Down‐regulation of LRRc17 secreted by BMSCs alleviates age‐related bone aging through autophagy enhancement
Authors: Liu Fei , Yuan Yujia , Li Yuanmin , Zhang Jie , Lu Yanrong , Chen Younan , Cheng Jingqiu 
Published: April 2020

BackgroundsThe impaired function of endogenous bone marrow mesenchymal stem cells (BMSCs) is a determinant in the development of osteoporosis (OP). BMSCs have been described the potential relevance of autophagy to the pathogenesis of OP. However, the underlying mechanism remains unclear. Our previous researches have found leucine‐rich repeat‐containing 17 (LRRc17) secreted by MSCs, a member of the LRR superfamily, was markedly enriched in the age‐related pathways. This study aims to investigate the effects of LRRc17 in aging process of MSCs.MethodsHydrogen peroxide (H2O2) induced aging MSCs and natural senescent cell cultivated from old and young C57BL/6 mice were used to evaluate the effects of LRRc17 on MSCs. Western blot, flow cytometry was used to detect autophagy, apoptosis and cell cycle respectively. Furthermore, mitochondrial function was measured by Mitosox, JC‐1, ATP. The mitochondrial morphology was detected by Mitotracker. Moreover, co‐culture was performed to detect the effects of LRRc17 secreted by MSCs on osteoblast (OB) and osteoclast (OC).ResultsMSCs exhibited a senescent phenotype after treatment with hydrogen peroxide. This result was improved after the addition of rapamycin. Interestingly, compared with the H2O2‐treated group, MSCs transfected with shLRRc17 showed increased DNA synthesis phase, and decreased apoptosis ratio, ROS (Fig.
1A, B, C). Autophagy related protein LC3 II, Beclin1 were up‐upregulated and P62 was down‐regulated (Fig.
1G). And mitochondrial morphology was also significantly improved (Fig.
1D, E). More importantly, LRRc17 reversed the loss of Ψm (Fig.
2A) and enhanced ATP content (Fig.
1F). Co‐culture showed that silenced LRRc17 in MSCs enhanced the osteogenic ability of OB and decreased the bone resorption capacity of OC, and the opposite result was obtained after overexpression (Fig.
2B).ConclusionsLRRc17 enhances characterizations of BMSCs through attenuating MSC senescence during OP, and which could lead to novel therapies to target altered bone formation.Support or Funding InformationThis work was supported by the Chengdu Giant Panda Breeding Research Foundation (2016cdspt001).Figure 1Figure 2
Co‐Delivery of Anti‐Inflammatory and Proliferative Agents by Injectable Hydrogel to Promote Tissue Repair after Acute Kidney Injury
Authors: Wang Yizhuo , Liu Shuyun , Zhao Meng , Wang Chengshi , Yuan Yujia , Li Lan , Liao Guangneng , Chen Younan , Cheng Jingqiu , Lu Yanrong , Liu Jingping 
Published: April 2020

ObjectIschemia‐reperfusion (I/R)‐induced acute renal injury has become a serious issue in clinical. The increasingly mortality, induced by chronic fibrosis and further renal dysfunction which caused by poor recovery of acute injury, has made it urgent to address kidney tissue repair. By using a renal I/R injury model, we aimed to investigate the role of a hydrogel‐based dual‐drug delivery platform on promoting tissue repair.MethodsAn injectable, self‐assembling peptide/heparin (SAP/Hep) hydrogel was employed to co‐deliver TNF‐α neutralizing antibody (anti‐TNF‐α) and hepatocyte growth factor (HGF). We analyzed the microstructure and controlled release properties of KLD2R/Hep hydrogel in vivo. The effects of the drug‐loaded hydrogel (SAP‐drug) on renal injury were evaluated in a rodent renal I/R injury model.ResultsIn vitro, the SAP/Hep hydrogel exhibited a faster release of anti‐TNF‐α with a sustained release of HGF, and both drugs could maintain better bioactivities after release. In vivo, combined anti‐TNF‐α/HGF showed better renal protective potential than anti‐TNF‐α or HGF alone. Compared with free‐drug (anti‐TNF‐α/HGF in solution) or SAP alone , SAP‐drug treatment significantly reduced the levels of serum creatinine (Scr)/blood urea nitrogen (BUN), tubular apoptosis, renal inflammatory factors and macrophage infiltration. Moreover, the SAP‐drug group exhibited better efficacy on promoting tubular cell proliferation and dedifferentiation than SAP or free‐drug alone, and thus reduced chronic renal fibrosis in I/R injury mice.ConclusionsWe confirmed that SAP could sequentially deliver anti‐TNF‐α and HGF to achieve anti‐inflammatory and pro‐proliferative effects simultaneously with only once injection, and thus can be a promising delivery platform for tissue repair.Support or Funding InformationThis work was supported by grants from National Natural Science Foundation of China (81571808, 31871001, 31200754) and Sichuan Science and Technology Program (2019YJ0069)Design of dual‐drug delivery KLD2R/Hep hydrogel for enhancing renal repair. The formation of the cationic KLD2R/anionic heparin hybrid hydrogel via electrostatic interaction, and heparin further had high affinity to HGF. The encapsulated anti‐TNF‐α and HGF were released from hydrogel in a sequential manner, and thus synergistically promoted tissue repair.Figure 1
Electrophysiological measurements of peripheral nerves in rats using the Qtrac approach
Authors: Pedersen Michael , Skov Martin Nors , Larsen Yrsa Marie Damgaard , Tankisi Hatice , Matchkov Vladimir 
Published: April 2020

IntroductionThe aim of this study was to build an implant and benchmark it for further improvement, specifically aimed to measure periphery nerves in rats. The nerve algorithm was based on the Qtrac approach for measurements of excitability measurements on motor and sensory nerves. This design was designed for enhanced measurements of nerve function related to diabetic neuropathy; a common and sometime a debilitating condition that eventually may result in foot ulcers and limb amputation. In order to better treat and prevent complications it is of great importance to recognize the sensory and autonomic dysfunction as early as possible. At the moment, it is unclear how and when the autonomic changes develop in relation to the somatosensory signs. The pathology of DN is primarily characterized by loss of nerve fibers. A typical course of the disease often starts with small‐fiber neuropathy/dysfunction in toes and feet, pain and paresthesia. Development of the proposed device addresses this problem.MethodsWe created a working pre‐amp and filter prototype and used conventional Qtrac algorithms. The performance consisted of multiple iterations to determine the best configuration for the amplification, signal‐to‐noise, filtration, space occupied on the PCB and battery use. The Qtrac package includes an automated stimulation<−>response threshold tracking software and hardware setup with specialized algorithms to determine multiple variables in nerve excitability studies. The device was measured in the nerves: sciatic, ulna and tail.ResultsEight rats were examined in 3 nerves (Fig
1A–1C). These data showed no significant difference between a regular Qtrac setup and the newly‐designed Qtrac setup. We showed a measurable difference between the nerves in question as shown in Figure
1D, showing 5 different variables obtained from the Qtrac protocol from the different nerves. The data is shown as a mean value of all rats in the study. At the same time, the difficulty in precisely measure in the same spot on different rats, became clear. Especially the Ulnar nerve proved too hard to use for this purpose, and we are reluctant to use this nerve in further studies.ConclusionThe study demonstrated a novel Qtrac design for measurements of periphery nerves in rats. The study revealed reliable data, showing significant difference in several excitability parameters for the three nerves. Potentially, explained by different amounts of fibers in the examined nerves, different fiber sizes and compositions. Further studies should include telemetric add‐on and measurements in rats suffering from DN. .needle placement in rats. A. Sciatic, B. Ulna, C. Tail. Graphs (D) show 5 different variables obtained from the Qtrac protocol from the different nerves.Figure 1
IQE.DYN‐Integrated Quantum Energy Dynamics by G.P. Einstein Physio‐Neuro Function
Authors: Sainvil Frantz , Tulp Orien Lee , Einstein George P. 
Published: April 2020

In a mathematical context, a dynamic system is a set of deterministic partial differential equations for the time neuro‐physiological processes with multiple variables. After long times a dynamic system displays one of a few possible types of dynamic states such as a steady state, oscillations or chaos. The dynamic equations typically depend on parameters that qualitatively influence the dynamic states of the biological system.The implications of the neuronal dynamics we studied experimentally and applied G. P Einstein Integrated Quantum Energy Dynamic of Physio‐Neuro Mathematical Function Model.G. P. Einstein Mathematical Model of Effective DYNAMICS Neuronal stimulation by Sigma Photonics Quantum Energy Therapy can be presented:
d ∑ IQBET/dt1 > d ∑ IQBPDE/dt2where: d ∑ I Q B E T ‐ derivative ∑ Integrated Quantum Bioelectromagnetic Energy Therapy d ∑ I Q B P D ‐ derivative ∑ Integrated QuantumIn neuronal stimulation, function is supported by Infrared Quantum Dynamics. It is the process of using an external source to activate ion channels causing depolarization of the neural membrane and evoking an action potential to propagate down the axon of a stimulated neuron.The Quantum Dynamics generate action potentials in both the central nervous system (CNS) and peripheral nervous system (PNS). Quantum Dynamics in Neuronal Stimulation by Sigma Photonics Quantum Energy was applied. The application of 11 basics mathematical function therapies is supported by specific integrated quanta energy matrixes. Each mathematical quantum energy function contains the specific matrix quanta energy therapy. By applying a controlled dose of Integrated Quantum Dynamics Physio‐Neuro stimulation by Each mathematical quantum energy function we can alter the properties of the tissue and the internal organs and bring their characteristics specific matrix quantum energy treatment. With careful choice of frequencies, which depend on each individual patient, his/her condition, degree of pathology and the time‐dependent properties of the treatment, we can treat a wide range of tumor cancers.The neuronal stimulation function was derived from neuro‐physio model as presented in figures
1 and
2.Support or Funding InformationSupported by Institutional Resources of the University of Science Arts & Technology [USAT], Montserrat and the Einstein Institute USA.Figure 1Figure 2
Changes in Steroids, Xenobiotics, and Drug‐Metabolism in Humans by Genetic Variations in NADPH Cytochrome P450 Oxidoreductase
Authors: Pandey Amit V. 
Published: April 2020

A broad spectrum of human diseases, including disordered steroid metabolism, are caused by mutations in the NADPH cytochrome P450 oxidoreductase (POR)
[1,
2]. Human POR is a diflavin reductase that transfers electrons from NADPH to small molecules, non‐P450 redox partners, and cytochrome P450 proteins in the endoplasmic reticulum
[2]. Cytochrome P450 proteins perform an extensive range of reactions, including the metabolism of steroids, drugs, and other xenobiotics. Therefore, genetic variations in POR change several metabolic pathways by changing the activities cytochromes P450
[2]. Due to this unique role of POR in metabolism, it was believed for a long time that genetic defects in POR are unlikely, and indeed a POR knockout mouse is embryonically lethal. However, in 2004, the first human patients with defects in POR were reported, and since then, over 200 different variations in POR have been found
[3,
4].Recently, we have characterized POR variations from non‐clinical samples
[5]. By analyzing the POR sequences from the 1000 genome and other sequencing projects, we identified potentially disease‐causing variations and characterized these by functional studies using recombinant proteins produced in bacteria, yeast, and mammalian cells
[6,
7].Identification of severe effects of POR mutations on both the drug and the steroid metabolizing cytochromes P450 indicates that likely pathogenic mutations may be found in non‐clinical population. Their combination as compound heterozygotes or homozygous may severely impact both steroid and drug metabolism by modifications of cytochrome P450 activities. Variations in POR need to be evaluated individually. Changes in drug and steroid metabolism due to genetic variations can be addressed using personalized metabolic profiling and supplementation using modified dosages of drugs and steroids.Support or Funding InformationSupported by grants from the Swiss National Science Foundation (31003A‐134926) & Novartis Foundation for Medical‐Biological Research (18A053) to AVP.
The Extent of Metaplastic Hard Tissue Within the Appendicular Skeleton of the Nine‐Banded Armadillo (Dasypus novemcinctus)
Authors: Heck Christian , Woodward Holly N. 
Published: April 2020

The developmental and functional advantages of vertebrate metaplastic hard tissue is still relatively unclear. Metaplastic tissue forms from the direct transformation of one cell type to another. Specifically, mineralization of preformed dense connective tissue results in intratendinous metaplastic tissue, and has been described in a variety of taxa and elements including turtle shell, reptilian osteoderms, sturgeon scutes, dinosaur cranial and vertebral elements, ossified tendons, portions of bird skeletons, and tendinous entheses in mammalian bone. However, histological analysis of the extent of metaplastic hard tissues, both across Mammalia and within individual mammalian bones, is lacking. Here we serially sectioned both tibiae of a nine‐banded armadillo, Dasypus novemcinctus [Xenarthra: Cingulata], transversely and longitudinally to identify the existence and possible patterns of metaplastic hard tissue. All skeletal elements were processed using standard procedure for the preparation of undecalcified bone. We found evidence of metaplastic hard tissue in the tibial crest, as well as the medial side of the diaphysis and the proximal lateral side of the tibia. The metaplastic tissue in the tibial crest appears interstitially within a primary woven bone matrix. Along the lateral side, metaplastic tissue forms a thin patch in the mid‐cortex superficial to compact coarse cancellous bone. The presence of metaplastic hard tissue seems consistent with tendinous insertions through fibrous entheses along the length of the tibia and tibial crest. The entrapment of metaplastic tissue within the tibia is likely due to appositional bone growth at the enthesis. However, the presence of metaplastic hard tissue on the medial side of the mid‐diaphyseal and proximal portion of the tibia does not appear to be associated with any tendinous insertions. Previously, unassociated metaplastic tissue was found extensively in osteoderms, ankylosaur tail clubs, and dinosaur cranial ornamentation such as the parietal frill of Triceratops. Study of metaplastic hard tissue in modern bone, independent of tendinous insertions, can improve our understanding of its potential biomechanical advantages and thereby possibly provide clues to its use in ornamentation and within the appendicular skeleton. Metaplastic hard tissues have likely been misidentified and understudied in previous investigations of modern hard tissue microstructure. Additionally, because Xenarthra is proposed to be one of the four original clades of placental mammals, analysis of metaplastic tissue in the skeleton of D. novemcinctus could have implications for early mammalian bone development patterns.Support or Funding InformationPartial funding was provided by Montana State University Undergraduate Scholar’s ProgramFigure 1
BDNF/TrkB Signaling Increases Autophagy Flux in Cervical Spinal Cord
Authors: Ducaji Emily C. , Pareja-Cajiao Miguel , Gransee Heather M. , Sieck Gary C. , Mantilla Carlos B. 
Published: April 2020

Autophagy is a homeostatic process for the elimination of damaged, unnecessary proteins or organelles within cells. Our overarching hypothesis is that autophagy contributes to neuromuscular dysfunction, specifically increased NMTF seen with aging. Understanding the mechanisms regulating autophagy flux is essential to discovering therapeutic agents which may mitigate the effects of neuromuscular dysfunction across conditions including old age. In recent studies, we found that in 18 month old mice autophagy in the cervical spinal cord is accelerated and neuromuscular transmission failure (NMTF) in diaphragm muscle‐phrenic nerve preparations is increased. Neurotrophin activity, specifically brain‐derived neurotrophic factor (BDNF) signaling through its high affinity receptor tropomyosin‐related kinase receptor subtype B (TrkB) decreases NMTF. Thus, we predict BDNF/TrkB signaling suppresses autophagy in phrenic motor neurons located in the cervical spinal cord. We hypothesize that inhibiting TrkB kinase activity accelerates autophagy in phrenic motor neurons. TrkBF616A mice at 18 months of age were randomized to receive 1 week treatment with vehicle (n=4) or 1NMPP1 (TrkB kinase inhibitor; n=6). Additionally, 18 month old wild type C57BL/6 mice (n=2) were studied. Whole cervical spinal cord (CSC) was harvested for measurements of autophagy marker protein expression analyzed by Western blotting. Data for the vehicle‐treated TrkBF616A and wild type mice was combined given lack of differences for any measurement. p62, a marker associated with the initiation of autophagy, was significantly decreased in the 1NMPP1‐treated group compared to vehicle‐treated or wild type mice. LC3, a protein marking the autophagosome membrane elongation phase, was not significantly changed across groups indicating that LC3‐tagged autophagosomes proceed to lysosomal degradation and do not accumulate. These results support our hypothesis that inhibiting BDNF/TrkB signaling accelerates autophagy flux in the CSC and reveal trophic effects of BDNF/TrkB signaling in the neuromuscular system. Accelerated autophagy may underlie age‐related neuromuscular dysfunction in a BDNF/TrkB signaling dependent manner.Support or Funding InformationSupport: NIH R01 AG057052
Is Chemotherapy‐Induced Cardiomyopathy Caused by Myocardial Ischemia?
Authors: Davidian Anahis , Mistry Maahi , Pucci Thomas , Kolz Christopher L , Shockling Lindsay , Enrick Molly , Yin Liya , Chilian William , Ohanyan Vahagn 
Published: April 2020

Over the last 50 years progress has been made in treating childhood and adult solid and hematological tumors. One commonly used chemotherapeutic agent is doxorubicin (DOX), an anthracycline antitumor antibiotic, which was discovered in early 1960s. Anthracycline (ANT) chemotherapy alone or in combination with other chemotherapeutic agents markedly improves the survival rate of cancer patients, particularly among children, where survival rates have more than doubled over 5 decades. However, the use of ANT chemotherapy is a double‐edged sword in that it can produce chemotherapy induced cardiac dysfunction (CRCD), which can cause death in 50% of the patients with who present with the cardiomyopathy. We hypothesized, that one of the causes for CRGD is a reduction in myocardial blood flow (MBF) that leads to cardiac myocyte hypoxia, producing small areas of ischemia and myocyte damage. We also hypothesize that increasing MBF by the coronary vasodilator chromonar prevents cardiomyocytes from hypoxic damage.To test these hypotheses, we treated mice genetically engineered to show cardiac myocyte fluorescence if the cells are hypoxic (hypoxia fate mapping [HFM]) with DOX for 6 weeks (group 1, N=8) and Dox + Chromonar for 6 weeks (group 2 N=8). Five weeks after treatment we started tamoxifen for 5 days to activate the HFM protocol. Tamoxifen induces Cre in cardiac myocytes and hypoxia activates a program that results in the expression of td‐tomato only in hypoxic cardiac myocytes. In tissue sections, hypoxic cardiac myocytes are visualized by fluorescence microscopy. Cardiac function was measured by ultrasound and MBF was measured with contrast echocardiography.Six weeks after DOX treatment many hypoxic myocytes were observed in the myocardium. Cardiac function was significantly decreased at the end of DOX treatment (% ejection fraction EF=39±3) compared to baseline (62±5.8%). Simultaneous administration of chromonar prevents the cardiomyocytes from hypoxic damage and prevents cardiac dysfunction development. %EF was 58±6 at the end of the chemotherapy. Chromonar also significantly increased MBF in group 2 mice compared to group 1 (P<0.05).Based on these data CRCD may be a type of ischemic heart disease where the treatment reduces MBF, which leads cardiomyocyte hypoxia, and causes progressive, accumulative cardiac injury. Moreover, increasing myocardial blood flow by chromonar prevents the cardiomyocytes from hypoxic damage and development of CRCD.Support or Funding InformationNIH, NHLBIPanel A shows the relationship between myocardial blood flow and cardiac work 6 weeks after DOX and DOX + Chromonar treatment. Panel B shows ejection fraction changes in DOX +Chromonar (Group 2 mice)Figure 1
Protein Kinase C Fusions Reveal Another Mechanism for Loss of Protein Kinase C Function in Cancer
Authors: Van An-Angela N. , Kunkel Maya T. , Baffi Timothy R. , Antal Corina E. , Newton Alexandra C. 
Published: April 2020

Over 60 protein kinase C (PKC) fusion genes have been identified in a wide variety of cancers, making PKC the most frequently fused AGC kinase. Our laboratory previously revealed that cancer‐associated point mutations annotated for PKC family members are generally loss‐of‐function, with functional studies revealing a tumor suppressive role for PKC. Here, we sought to determine whether PKC fusions may also be loss‐of‐function. In our analysis, we characterized TANC2‐PRKCA, a fusion identified in lung squamous cell carcinoma that retains the catalytic domain of PKCα, and PRKCA‐CDH8, a fusion identified in breast cancer that retains the regulatory domain of PKCα. Overexpression of TANC2‐PKCα in cells showed that the fusion protein is constitutively active; however, it lacks phosphorylation at key sites required for the stability of the enzyme. Most importantly, without its amino‐terminal pseudosubstrate, the fusion protein is unable to adopt the autoinhibited, stable conformation of wild‐type PKC, rendering it markedly unstable. To determine whether the fusion protein could accumulate in cells given this instability, we utilized CRISPR/Cas9‐mediated gene editing to express TANC2‐PRKCA. Indeed, while the fusion mRNA was detected in the CRISPR‐edited clones, we were unable to detect the fusion protein. Thus, although the fusion is rendered constitutively active through loss of autoinhibition, its subsequent instability prevents its accumulation in cells, making it paradoxically loss‐of‐function. In contrast to fusions that retain the PKC catalytic domain, fusions that retain the PKC regulatory domain are intrinsically loss‐of‐function with respect to PKC activity. However, we were interested in exploring whether regulatory domain fusions could also act in a dominant‐negative manner. Overexpression of PKCα‐CDH8 in cells suppressed endogenous PKC activity, both basally and following agonist stimulation. This suggests that PKC regulatory fusions are not only loss‐of‐function by loss of the catalytic domain, but also dominant negative by competition of the ligand‐sensing modules for binding the allosteric activator diacylglycerol. Taken together, our data demonstrate that both catalytic and regulatory domain PKC fusions are loss‐of‐function, revealing another mechanism by which PKC activity is lost in cancer and supporting a tumor suppressive role for PKC.Support or Funding InformationThis work was supported by NIH R35 GM122523 (ACN). ANV, TRB, and CEA were supported by the UCSD Graduate Training Program in Cellular and Molecular Pharmacology (T32 GM007752), and CEA was supported by an NSF Graduate Research Fellowship (DGE1144086).