bluuebunny/crossref_metadata_2025_split huggingface dataset:
Zoledronic Acid Activates TRPV1 Channels: Possible Role in Cell Proliferation and Pain
Authors: Scala Rosa , Maqoud Fatima , Latorre Ramon , Scilimati Antonio , Tricarico Domenico 
Published: April 2020

Bisphosphonates (BPs) are the most used bone specific anti‐resorptive drugs. Lately there is an increasing interest in BPs capability to prolong osteoblasts lifespan and mineralize; however, the mechanism of action which underpin these effects are nowadays poorly understood. In addition, an increasing concern about BPs pain‐relieving effects is now emerging. Empirical evidence showed BPs may exert anti‐nociceptive, anti‐hyperalgesia and anti‐allodynic properties against multiple stimuli; targets involved in these analgesic effects remains elusive. FDA alert showed that the chronic use of BPs is linked with an increased risk of musculoskeletal pain; mechanisms underlying this sensation are not known. TRPV1 channel is actually recognized to play a key role in bone formation process; as well, it appeared to be involved in bone pain sensation. Capsaicin, a selective agonist of TRPV1 channel, is used for treating different bone pain conditions. We have recently demonstrated that the BP Zoledronic acid (ZOL) can activate TRPV1 currents on osteoblasts. In the mineralization assay, the co‐application of ZOL with capsazepine, an antagonist of TRPV1 ion channel, reduced ZOL‐induced mineralization (Scala et al., Cancer 2019). We tested ZOL effects on TRPV1 ion channel in differentiated neuronal SH‐SY5Y cells and in HEK293 cells transfected with cDNA codifying for WT and PI(4,5)P2‐binding site mutant TRPV1 ion channels. In whole‐cell patch clamp experiments on neuronal SH‐SY5Y cells, ZOL (100μM) activated strong outward currents both at negative and positive voltages. At positive membrane voltages ZOL (100μM)‐ evoked currents were closed after the application of capsazepine (1μM). On WT TRPV1 channel expressed on HEK293 cells, ZOL (100 μM) activates currents by 37% at −60 mV, by +81% at +60 mV and +80% at +180 mV (Vm). As well, on K694A mutant, ZOL was able to activate currents by +10%, +63% and +76% at −60 mV, +60 mV and +180 mV (Vm), respectively. On the contrary, no activation of currents was observed on the R579A mutant.The ZOL‐induced activation of TRPV1 channel contributes to the mineralization of osteoblasts and to the pain‐relieving effect of ZOL in neurones. Experiments on mutant TRPV1 ion channel suggest that the arginine in the S4–S5 linker of the channel is essential for the drug interaction.Support or Funding InformationThis research was funded by Reg. Puglia (Italy) project “Cluster in Bioimaging” code QZYCUM0 D.T and A.S., and Fondecyt grants 1150273, 1190293 R.L.. R.S. was supported by GLOBAL THESIS Study (2016–2017) by Univ. degli Studi di Bari “Aldo Moro” at CINV “Centro Interdisciplinario De Neurociencia De Valparaíso”.
IgM Antibodies with atheroprotective properties in heart transplants are IgM anti‐phosphorylcholine
Authors: Labarrere Carlos Alberto , Hardin James W. , Jaeger Beate R. , Bhatt Deepak L. , Kassab Ghassan S. 
Published: April 2020

Introduction and ObjectiveAlthough the etiology of transplant coronary atherosclerosis (TCA), the major cause of long‐term morbidity and mortality in heart transplantation (tx), is unknown, several immunological and non‐immunological causes (innate immunity, inflammation, cell activation, coagulation) have been proposed. Our group and others found that high circulating levels of C‐reactive protein (CRP), a major soluble pattern recognition receptors (sPRR) in humans, associate with development and progression of native atherosclerosis and TCA; although the pathogenic role of different isotypes of CRP (native, non‐native and proinflammatory monomeric) in atheroprotection or atherogenesis remains unclear. Immunoglobulin (Ig) M natural antibodies (NAbs), another innate sPRR, conveys an atheroprotective function, since high titers of IgM anti‐phosphorylcholine (IgM anti‐PC) NAbs associate with reduced atherosclerosis. Our objective was to determine the protective role of sPRR, more specifically IgM anti‐PC, in TCA.MethodsAdult heart transplant patients (n=172; 5.2±1.0 serial biopsies/patient in first 3‐months post‐tx) followed 8.9±5.0 years were studied. CAV and CAV severity were evaluated with angiograms (5.3±1.0/patient). Percentage of microvascular IgM was calculated from double staining (IgM/CD31 or Ulex europaeus lectin). IgM and IgM anti‐ PC from biopsy eluates were evaluated with immunoblotting and ELISA. Analyses used logistic regression; and Kaplan‐Meier curves for different tertile groups.ResultsPatients had low (<30% positive microvessels in all biopsies; n=67), high (>50% positive microvessels in all biopsies; n=64) or oscillatory IgM (IgMosc; low and high in all biopsies; n=41). Steady high IgM had an atheroprotective effect at 1‐, 5‐ and 10‐years (p<0.001) post‐tx compared with steady low IgM; while IgMosc was only atheroprotective at 5‐years (p<0.04). To determine if IgM was bound to PC, we incubated biopsies with high IgM (n=5) with PC, CRP or albumin, and immunoblots from eluates showed that myocardial IgM was removed by PC and CRP but not albumin. ELISA studies of biopsy eluates confirmed IgM anti‐PC following PC (56.7±20.0 U/ml) and CRP (60.5±18.3 U/ml), but not albumin (0.0±0.0 U/ml) incubation. Immunohistochemistry of biopsies post‐elution showed a complete absence of IgM.ConclusionsAtheroprotective IgM antibodies are NAbs to PC and future research needs to demonstrate if enhancing IgM anti‐PC NAbs ameliorates TCA, and if native CRP enhances atheroprotection.
High Expression of F11R/JAM‐A is Predominant in Triple‐negative Breast Cancer Subtype and Correlates with Poor Prognosis in Breast Cancer Patients
Authors: Fang Chih-Yeu , Chen Chi-Long , Hsiao Michael 
Published: April 2020

Breast cancer (BrC) is the leading type of cancer in women, accounting for 25% of all female tumor cases. It affects about 13% of women worldwide and is the most common cause of cancer‐related death in women. Breast cancer is a heterogeneous disease that contains several subtypes with substantial differences in pathology and clinical outcome. F11 receptor (F11R), also known as the junctional adhesion molecules‐A (JAM‐A), is one of the members of an immunoglobulin superfamily of cell adhesion receptors. F11R is a component of tight junction molecules and involved in a variety of physiological processes including cell polarity, cell morphology and leukocyte migration. Overexpression of F11R has been reported in BrC, but its correlation with clinical prognosis is controversial. In this study, we investigated and clarified the role of F11R in breast carcinogenesis. Two BrC cohorts with a total of 542 cases were used to confirm the correlation of F11R expression and the clinical outcome. High expression of F11R was found to be associated with tumor size (p=0.035), grading (p=0.001), and recurrence (p=0.001) in BrC. Cox model analysis revealed that high expression of F11R is significantly associated with poor overall and disease‐free survival in BrC patients. Interestingly, high expression of F11R is predominant in triple‐negative breast cancer (TNBC, >65%) subtype and is inversely correlated with the expression of estrogen and progesterone receptors. Our data validated the expression of F11R in BrC, suggesting that it is a valuable biomarker for prognostic prediction of recurrence and survival, especially for TNBC.Support or Funding InformationThis research was supported by Academia Sinica [AS‐SUMMIT‐108].
Contribution of the ventral tegmental area to spontaneous coactivation of the motor and cardiovascular systems in decerebrate rats
Authors: Ishii Kei , Asahara Ryota , Komine Hidehiko , Liang Nan , Matsukawa Kanji 
Published: April 2020

Central command is a feedforward signal from higher brain centers, regulating the cardiovascular system in association with exercise. Although the concept is widely accepted, neural circuits responsible for central command were still unclear. We hypothesized that the ventral tegmental area (VTA) contributes to generating spontaneous motor activity and concomitant cardiovascular responses. To test the hypothesis, a GABAergic receptor agonist (muscimol) and saline were injected into the bilateral VTA in 9 decerebrate paralyzed rats. In the rats, spontaneous motor activity occurred at frequency of 80 ± 15 times/h (mean ± SE) and accompanied increases in heart rate (by 11 ± 2 beats/min) and mean arterial blood pressure (MAP, by 36 ± 4 mmHg). Muscimol into the VTA reduced the frequency of spontaneous motor activity (19 ± 8 times/h) and the baseline MAP, but not HR. The concomitant tachycardiac and pressor responses (by 12 ± 3 beats/min and 29 ± 3 mmHg) were similar (P > 0.05) before and after the muscimol injection. In contrast, saline into the VTA had no effects (P > 0.05) on the frequency of spontaneous motor activity and the baseline and peak values of HR and MAP. These findings suggest that the VTA plays a significant role in the synchronized activation of the motor and cardiovascular systems in decerebrate rats.Support or Funding InformationSupport of this work was provided by Grant‐in‐Aid for Research Activity Start‐up (grant number 17H07393) from the Japan Society for the Promotion of Science.
Diabetic metabolic dysregulation and chronic kidney disease induce specific perturbations in coronary microvascular function in swine
Authors: Sorop Oana , Van De Wouw Jens , Heinonen Ilkka , Merkus Daphne , Danser Jan A.H. , Duncker Dirk J. 
Published: April 2020

IntroductionComorbidities such as diabetes mellitus (DM), hypercholesterolemia (HC) and chronic kidney disease (CKD), are often associated with altered coronary microvascular function. However, the specific mechanisms affected by the various comorbidities have not been investigated in detail. Here we studied endothelium‐dependent and ‐independent vasodilation of isolated small coronary arteries, as well as vasoconstrictor responses to endothelin‐1 and their specific mechanisms of action in swine with multiple comorbidities.Methods and ResultsDM (streptozotocin, 150mg/kg i.v.) and hypercholesterolemia (high fat diet) were induced in 10 female swine, which were followed for 6 months (DM+HC). In a separate group of 9 female swine, CKD (by renal artery embolization) was added on top of the two risk factors and animals were followed for 6 months (DM+HC+CKD). 12 female healthy swine on normal pig‐chow served as controls. At sacrifice, coronary small arteries were isolated and vasomotor control was studied in vitro. Both DM+HC and DM+HC+CKD groups showed impaired endothelium‐dependent vasodilation to bradykinin, mainly via loss of EDHF‐dependent mechanism. No changes were seen in the response to the exogenous NO‐donor, SNAP, indicating preserved smooth muscle cell function. Coronary arteries from swine with DM+HC exhibited a blunted vasoconstriction to endothelin‐1, (ET‐1), due to increased ETB‐mediated vasodilation. In contrast, adding CKD to DM+HC resulted in enhanced ET‐1 vasoconstriction.ConclusionDiabetic metabolic dysregulation and chronic kidney disease induce comorbidityspecific perturbations in coronary microvascular function in swine.Support or Funding InformationSupported by grants from the European Commission FP7‐Health‐2010 grant MEDIA‐261409, the Netherlands CardioVascular Research Initiative: an initiative with support of the Dutch Heart Foundation [CVON2014‐11(RECONNECT)] and The Academy of Finland 251272.
G protein βγ translocation to the Golgi activates the mitogen‐activated protein kinases via phosphoinositide 3‐kinase γ
Authors: Khater Mostafa Kamel , Wei Zhe , Huang Wei , Lokeshwar Bal L. , Lambert Nevin A. , Wu Guangyu 
Published: April 2020

The activation of G protein‐coupled receptors (GPCRs) induces the translocation of G protein βγ complex from the plasma membrane (PM). However, its functions are not well known. Here we demonstrated that among the entire twelve Gγ family members, Gγ9 is the most translocatable Gγ subunit. More than 80% of Gγ9, in complex with Gβ1, relocated from the PM to the Golgi in response to CXCR4 activation by stromal cell‐derived factor 1α (SDF1α) and the half time of the translocation was about 5 seconds. To study the function of Gβγ translocation to the Golgi, we focused on the activation of the mitogen‐activated protein kinases (MAPK) ERK1/2. CRISPR‐Cas9‐mediated knockout of Gγ9 abolished ERK1/2 activation by SDF1α, whereas knockout of the least‐translocatable Gγ3 had no effect. To investigate if Gβγ‐mediated ERK1/2 activation indeed occurred on the Golgi, we employed an inducible system to specifically express Gβγ onto the Golgi. Interestingly, the targeted expression of Gγ2, Gγ3 and Gγ9, in complex with Gβ1, onto the Golgi each induced remarkable ERK1/2 activation. Such constitutive ERK1/2 activation was blocked by the expression of Gα subunits, the targeted expression of the Gβγ blocker GRK2ct at the Golgi, and chemical treatments to disrupt the Golgi structure. By contrast, the expression of individual Gγ subunits at the PM did not activate ERK1/2. To define the downstream effectors of Gβγ, we determined the effect of selective pharmacological inhibition of phosphoinositide 3‐kinase (PI3K), phospholipase C, PKC and PKD on ERK1/2 activation. This strategy identified PI3Kγ and PI3Kδ to be essential for Gβγ‐mediated ERK1/2 activation. Indeed, knockout of PI3Kγ by CRISPR‐Cas9 almost abolished ERK1/2 activation induced by both SDF1α and Gγ9 targeting to the Golgi. Similar to CXCR4, α2‐adrenergic receptor‐mediated ERK1/2 activation was also dramatically inhibited by Gγ9 and PI3Kγ knockout. We further showed that knockout of Gγ9 and PI3Kγ markedly suppressed the migration and invasion of prostate cancer PC3 cells in response to SDF1α stimulation in vitro. Gγ9 and PI3Kγ knockout also significantly reduced the bone metastasis of PC3 tumors following intra‐cardiac injection of PC3‐Luc cells in athymic nude mice. Overall, our data demonstrate a novel function for Gβγ translocation to the Golgi to spatiotemporally control the activation of the MAPK pathway by GPCRs and tumor progression.Support or Funding InformationNIH R01 GM118915
Compensatory Plasticity of Breathing in Patients with Neuromuscular Disease
Authors: Smith Barbara K 
Published: April 2020

Neuromuscular diseases are comprised of variable, often inherited conditions and characterized by muscular (e.g. Duchenne muscular dystrophy), neural (e.g. amyotrophic lateral sclerosis) or a mixed neuromuscular (e.g. Pompe disease) dysfunction. While individual neuromuscular disorders differ in the underlying mechanism and exhibit a unique disease presentation, the consistently leading cause of morbidity and mortality is respiratory failure. Loss of breathing function is literally a “life‐or‐death" issue for patients. Clinical management of breathing relies upon pulmonary function tests to estimate the degree of respiratory involvement in patients, but standard pulmonary function test results can be biased by learning, cognition, or poor effort. This presentation will discuss compensatory neuromuscular adaptations to preserve breathing in the face of worsening disease, including changes in neural drive, neuromuscular junction plasticity, and a greater reliance on less‐affected respiratory motor pools. Detection of these adaptations is needed for prescribing supportive therapies with greater precision and identifying clinically meaningful changes in response to treatments. Citing natural history studies from our team and others, this presentation will feature clinical approaches to objectively evaluate respiratory motor control and gain mechanistic insights into neuromuscular disease. The control of breathing will then be discussed in the context of clinical trials, with an emphasis on current and emerging treatments directed toward improvements in ventilation. Examples will highlight basic‐clinical collaborations that promote new therapeutic frontiers for patients with neuromuscular disease.Support or Funding InformationThis work is supported by National Institutes of Health awards R21 HD090752 and U01 HL121842.
Role of TMAO, a Gut Metabolite, on Proline‐Rich Proteins
Authors: Kumari Kritika , Singh Laishram Rajendrakumar 
Published: April 2020

Osmoprotectants, a group of low molecular weight compounds, are accumulated in various cells to protect organism from various stresses. Most of the osmoprotectants (osmolytes) are known to stabilize proteins by assisting them in proper folding into the native conformation. However, Trimethylamine N‐oxide (TMAO), a human gut metabolite, is one such osmolyte that is believed to have both stabilizing and destabilizing effect on different proteins. Till date the destabilizing effect of TMAO has been confined only to the fast‐folding proteins. We, therefore, investigated the destabilizing effect of TMAO on folding pattern of various slow‐folding proteins.MethodologyFolding pattern of chosen proteins in absence and presence of TMAO were studied using enzyme kinetic assays. Different spectroscopic probes were further used to measure effect on native and refolded state structure. Light Scattering measurements and electron micrographs were obtained using Dynamic light scattering (DLS) and Transmission electron microscopy (TEM) respectively to measure aggregation status of different protein species.ResultsProtein stability and enzyme activity studies revealed TMAO acts as a denaturant. Among all osmolytes this effect was found to be unique to TMAO only. No recovery of the enzyme activity of refolded protein even after prolonged incubation without formation of aggregates confirms TMAO interfere with refolding of proteins with high number of proline residues. Structural studies revealed refolding of protein to a compact denatured state. Changed behavior of protein from denatured to native state was observed upon addition of Peptidyl‐Prolyl Isomerase (PPIase).ConclusionWe discovered that TMAO fails to stabilize or refold the slow folding proteins. Furthermore, we showed that in proteins with high number of proline residues, proline cis‐trans isomerization becomes the rate limiting step in protein folding process, thus resulting into slow folding of proteins. Recovery of native like activity of protein in presence of PPIase indicate that TMAO might have interfered cis‐trans conversion generating a non‐native protein species. This study highlights the potential role of TMAO in protein folding and associated pathological consequences.
To investigate whether metastasis‐related mechanism are the same in different organs through the in vivo selection of colorectal cancer cell lines from liver and lung metastases
Authors: Hsu Tai-I , Hsiao Micheal 
Published: April 2020

The mortality and incidence rate of colorectal cancer is currently one of the top three cancers in Taiwan. In the United States the mortality and incident rate of colorectal cancer also the one of top three cancers. The colorectal cancer can use the stool occult blood examination as the early screening tool to achieve the early prevention. Although, the non‐invasive and highly effective method of early prevention, there still have some patients who cannot be screened or who are false negatives may have had micrometastasis or some treatment difficulties when the disease is discovered. The current therapeutic strategies are surgery, radiation, chemotherapy, chemotherapy concurrent radiotherapy and target therapy. The survival rate of patients is improved within the advanced medical technology and drug. However, for metastatic colorectal cancer, it is still unable to effectively improve the survival rate. The metastasis is also the majority cause of death in cancer. In our proposal, the genetic profile of metastatic colorectal cancer will be analyzed and the drug efficacy will be verified in metastatic cancer cells. The xerography model had been established. The cancer cells can be injected on the subserosal of cecum wall. In the in vivo tracking, the mice used the IVIS to tracking the immunofluorescence signal. After 4 weeks the signal can be detected in the other organ such as liver and lung. To confirm the signal from the metastasis organs, the mice were sacrificed and collected the organ to confirm the cells metastatic to liver and lung. However the cells in the transwell assay seem can not show the migration or metastatic function. The IHC staining for human mitochondria cells verified the cells with large ratio of nucleus and substance. The optical observation did not show the difference. The experimental assay showed the control group did not show highly growth rate. The cell growth rate in lung group is increased compared with the other groups. In the cancer progression theory, the cancer stemness theory has been mentioned. The western blotting assay related of stemness theory showed that lung group with dominates notch1 level decreased. Therefore the non‐coding RNA array was a tool for us to screen the candidate non‐coding RNA included 4 up‐regulation and 1 down‐regulation miRNAs. In the future, the miRNAs will be the target to investigated.Support or Funding InformationThis research was supported by Academia Sinica [AS‐SUMMIT‐108].
Molecular and cellular analysis of intestinal lactase‐phlorizin hydrolase gene variants unravel a heterogeneous pathogenic pattern of congenital lactase deficiency
Authors: Marten Lara M. , Wanes Dalanda , Santer René , Naim Hassan Y. 
Published: April 2020

Congenital lactase deficiency (CLD) is a rare autosomal recessive genetic disorder, leading to a dysfunctional lactase‐phlorizin hydrolase (LPH). LPH is a ß‐galactosidase located in the intestinal brush border membrane and indispensable for lactose digestion. Impaired LPH‐function results in severe symptoms within a few days after birth with the onset of nursing. The symptoms include watery diarrhea and consequently dehydration, leading to potential life‐threatening disease of the newborn. Several mutations in the coding region of LPH have been characterized, occurring in homozygote or compound heterozygote patterns of inheritance. The basis of the genotype‐phenotype relationship in CLD on a molecular and cellular level has not been studied in details. Here we analyze the influence of the mutations p.S688P, p.E1612X, p.S1121L, that have been identified in patients from different ethnic groups, on the structure, trafficking and function of the LPH protein. For this purpose, LPH‐mutants were generated by site‐directed mutagenesis and transiently expressed in COS‐1 cells. Additionally we concluded studies regarding the interaction of these mutants with the wild‐type‐LPH protein. The results reveal phenotypic variations in the protein trafficking of the mutants between the ER and the Golgi. Additionally, the LPH‐mutant harboring the mutation p.S688P showed residual lactose digesting capacity. These variations implicate possible clinical correlation as for example different course and severity of disease. These data are relevant for the further understanding of CLD, potential impact for heterozygote mutation carriers and possible invention of treatment strategies, as for example protein stabilization through chaperones.Support or Funding InformationIntramural funds, University of Veterinary Medicine Hannover