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George R R Bell, Dean E Natwick, Sean R Collins
Neutrophils are key early responders in the innate immune response that use chemotaxis, the directed migration along chemical gradients, to reach sites of infection or inflammation. This process requires integrating inputs from cell surface receptors with the cell's polarity and motility signaling network, in which highly dynamic and interconnected signaling by Rho-family GTPases plays a central role. To understand this fundamentally important behavior, we describe a high-resolution, under-agarose chemotaxis assay for use with neutrophil-like cell lines (HL-60 or PLB-985) or with primary neutrophils...
2018: Methods in Molecular Biology
Tatiana M Vinogradova, Syevda Tagirova Sirenko, Edward G Lakatta
Spontaneous beating of the heart pacemaker, the sinoatrial node, is generated by sinoatrial node cells (SANC) and caused by gradual change of the membrane potential called diastolic depolarization (DD). Submembrane local Ca2+ releases (LCR) from sarcoplasmic reticulum (SR) occur during late DD and activate an inward Na⁺/Ca2+ exchange current, which accelerates the DD rate leading to earlier occurrence of an action potential. A comparison of intrinsic SR Ca2+ cycling revealed that, at similar physiological Ca2+ concentrations, LCRs are large and rhythmic in permeabilized SANC, but small and random in permeabilized ventricular myocytes (VM)...
July 25, 2018: International Journal of Molecular Sciences
Antonio F Melo Junior, Polyana L M Dalpiaz, Glauciene J Sousa, Phablo Wendell C Oliveira, Antônio M Birocale, Tadeu U Andrade, Gláucia R Abreu, Nazaré S Bissoli
AIMS: Hypertension is a highly prevalent disease that has been correlated to severe organ damage and mortality. However, the role of androgens in hypertension is controversial. The aim of this study was to evaluate the cardiac effects of the nandrolone decanoate (NDL) in male SHR. MAIN METHODS: At 12 weeks of age, male SHR rats were separated into three groups: Control (CON), Nandrolone 10 mg/kg twice weekly (NDL), and NDL plus Enalapril 10 mg/kg/day (NDL-E) groups...
July 21, 2018: Life Sciences
Bo Sun, Jinhong Wei, Xiaowei Zhong, Wenting Guo, Jinjing Yao, Ruiwu Wang, Alexander Vallmitjana, Raul Benitez, Leif Hove-Madsen, S R Wayne Chen
Sarcoplasmic reticulum (SR) Ca2+ cycling is governed by the cardiac ryanodine receptor (RyR2) and SR Ca2+-ATPase (SERCA2a). Abnormal SR Ca2+ cycling is thought to be the primary cause of Ca2+ alternans that can elicit ventricular arrhythmias and sudden cardiac arrest. Although alterations in either RyR2 or SERCA2a function are expected to affect SR Ca2+ cycling, whether and to what extent altered RyR2 or SERCA2a function affects Ca2+ alternans is unclear. Here we employed a gain-of-function RyR2 variant (R4496C) and the phospholamban-knockout (PLB-KO) mouse model to assess the effect of genetically enhanced RyR2 or SERCA2a function on Ca2+ alternans...
July 9, 2018: Journal of Biological Chemistry
Patricia Munsch-Alatossava, Reijo Käkelä, Dominique Ibarra, Mohammed Youbi-Idrissi, Tapani Alatossava
Cold storage aims to preserve the quality and safety of raw milk from farms to dairies; unfortunately, low temperatures also promote the growth of psychrotrophic bacteria, some of which produce heat-stable enzymes that cause spoilage of milk or dairy products. Previously, N2 gas flushing of raw milk has demonstrated significant potential as a method to hinder bacterial growth at both laboratory and pilot plant scales. Using a mass spectrometry-based lipidomics approach, we examined the impact of cold storage [at 6°C for up to 7 days, the control condition (C)], on the relative amounts of major phospholipids (phosphatidylethanolamine/PE, phosphatidylcholine/PC, phosphatidylserine/PS, phosphatidylinositol/PI, and sphingomyelin/SM) in three bovine raw milk samples, and compared it to the condition that received additional N2 gas flushing (N)...
2018: Frontiers in Microbiology
Fei Hong, Huan-Xin Zhang, Chong Chen, Zhi-Ling Yan, Qing-Yun Wu, Ling-Yu Zeng, Zheng-Yu Li, Kai-Lin Xu
OBJECTIVE: To investigate the effect of steadily down-regulating the expression of VE-cadherin on the chemotheraputic sensitivity of K562 cells, and explore its possible mechanism. METHODS: Specifically targeting interference sequences carrying human VE-cadherin were designed, the recombinant lentiviral vector containing the IRES-GFP and NEO segment was constructed; recombinant lentivirus was generated by three-plasmids packing system, and transfected into K562 cells, then the cells steadily down-regulated were sorted...
June 2018: Zhongguo Shi Yan Xue Ye Xue za Zhi
Eli Fernández-de Gortari, L Michel Espinoza-Fonseca
Sarcoplasmic reticulum Ca2+ -ATPase (SERCA) is critical for cardiac Ca2+ transport. Reversal of phospholamban (PLB)-mediated SERCA inhibition by saturating Ca2+ conditions operates as a physiological rheostat to reactivate SERCA function in the absence of PLB phosphorylation. Here, we performed extensive atomistic molecular dynamics simulations to probe the structural mechanism of this process. Simulation of the inhibitory complex at super-physiological Ca2+ concentrations ([Ca2+ ] = 10 mM) revealed that Ca2+ ions interact primarily with SERCA and the lipid headgroups, but not with PLB's cytosolic domain or the cytosolic side of the SERCA-PLB interface...
June 22, 2018: Journal of Biological Chemistry
Leonardo Bocchi, Monia Savi, Valeria Naponelli, Rocchina Vilella, Gianluca Sgarbi, Alessandra Baracca, Giancarlo Solaini, Saverio Bettuzzi, Federica Rizzi, Donatella Stilli
BACKGROUND/AIMS: Dietary polyphenols from green tea have been shown to possess cardio-protective activities in different experimental models of heart diseases and age-related ventricular dysfunction. The present study was aimed at evaluating whether long term in vivo administration of green tea extracts (GTE), can exert positive effects on the normal heart, with focus on the underlying mechanisms. METHODS: The study population consisted of 20 male adult Wistar rats...
2018: Cellular Physiology and Biochemistry
Tatiana M Vinogradova, Syevda Sirenko, Yevgeniya O Lukyanenko, Dongmei Yang, Kirill V Tarasov, Alexey E Lyashkov, Nevin J Varghese, Yue Li, Khalid Chakir, Bruce Ziman, Edward G Lakatta
BACKGROUND: Spontaneous firing of sinoatrial node cells (SANCs) is regulated by cAMP-mediated, PKA (protein kinase A)-dependent (cAMP/PKA) local subsarcolemmal Ca2+ releases (LCRs) from RyRs (ryanodine receptors). LCRs occur during diastolic depolarization and activate an inward Na+ /Ca2+ exchange current that accelerates diastolic depolarization rate prompting the next action potential. PDEs (phosphodiesterases) regulate cAMP-mediated signaling; PDE3/PDE4 represent major PDE activities in SANC, but how they modulate LCRs and basal spontaneous SANC firing remains unknown...
June 2018: Circulation. Arrhythmia and Electrophysiology
Peter D Martin, Zachary M James, David D Thomas
We have used site-directed spin labeling and electron paramagnetic resonance (EPR) to map interactions between the transmembrane (TM) domains of the sarcoplasmic reticulum Ca2+ -ATPase (SERCA) and phospholamban (PLB) as affected by PLB phosphorylation. In the cardiac sarcoplasmic reticulum, PLB binding to SERCA results in Ca-dependent enzyme inhibition, which is reversed by PLB phosphorylation at Ser16. Previous spectroscopic studies on SERCA-PLB have largely focused on the cytoplasmic domain of PLB, showing that phosphorylation induces a structural shift in this domain relative to SERCA...
June 5, 2018: Biophysical Journal
Ian Warbrick, Simon W Rabkin
Heart failure with preserved ejection fraction (HFpEF) represents an important cardiac condition because of its increasing prevalence, resistance to treatment and high associated morbidity and mortality. Two of the major mechanisms responsible for HFpEF are impaired cardiomyocyte sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a), which is responsible for calcium reuptake into the SR, and cardiac fibroblasts/myofibroblasts that produce collagen or myocardial fibrosis. Phospholamban (PLB), in the SR and endoplasmic reticulum, is the primary regulator of SERCA2a in the heart and acts as a reversible inhibitor of SERCA2a...
May 25, 2018: Peptides
Andressa Heimbecher Soares, Nidia Celeste Horie, Lucas Augusto Piccinin Chiang, Bruno Caramelli, Mariana Gomes Matheus, Alexandre Holthausen Campos, Luciana Cavalheiro Marti, Fernanda Agostini Rocha, Marcio C Mancini, Elaine Maria Frade Costa, Cintia Cercato
BACKGROUND: Obesity causes secondary hypogonadism (HG) in men. Standard testosterone (T) replacement therapy improves metabolic parameters but leads to infertility. OBJECTIVE: To evaluate clomiphene citrate (CC) treatment of adult men with male obesity-associated secondary hypogonadism (MOSH). DESIGN: Single-center, randomized, double-blind, placebo-controlled trial. PARTICIPANTS: Seventy-eight men aged 36.5 ± 7...
June 2018: International Journal of Obesity: Journal of the International Association for the Study of Obesity
Amin Sheikh, Kai Y Chau, Helen M Evans
Histological abnormalities, including chronic hepatitis, fibrosis, and steatosis, are increasingly reported in liver biopsies of children after LT. These changes may be progressive and represent a form of rejection. Liver biochemistry is often initially normal. Our LT program began in 2002, utilizing tacrolimus and low-dose steroids for the first year post-LT. Patients undergo a protocol biopsy at 1 year post-LT prior to stopping steroids, then at 5 years and every 5 years thereafter. Target tacrolimus levels are 5-8 μg/L and 3-5 μg/L after 3 and 12 months, respectively...
August 2018: Pediatric Transplantation
Dolores Montalvo, Perla Pérez-Treviño, Katheryne Madrazo-Aguirre, Fabio A González-Mondellini, Hipólito O Miranda-Roblero, Diego Ramonfaur-Gracia, Mariana Jacobo-Antonio, Maritza Mayorga-Luna, Norma L Gómez-Víquez, Noemí García, Julio Altamirano
Hypothyroidism (Hypo) is a risk factor for cardiovascular diseases, including heart failure. Hypo rapidly induces Ca2+ mishandling and contractile dysfunction (CD), as well as atrophy and ventricular myocytes (VM) remodeling. Hypo decreases SERCA-to-phospholamban ratio (SERCA/PLB), and thereby contributes to CD. Nevertheless, detailed spatial and temporal Ca2+ cycling characterization in VM is missing, and contribution of other structural and functional changes to the mechanism underlying Ca2+ mishandling and CD, as transverse tubules (T-T) remodeling, mitochondrial density (Dmit ) and energy availability, is unclear...
June 2018: Cell Calcium
Hsin-Yuan Fang, Meng-Yu Hung, Yueh-Min Lin, Sudhir Pandey, Chia-Chien Chang, Kuan-Ho Lin, Chia-Yao Shen, Vijaya Padma Viswanadha, Wei-Wen Kuo, Chih-Yang Huang
Earlier studies have shown that estrogen possess protective function against the development of pathological cardiac hypertrophy. However, the molecular mechanisms of estrogens (E2) protective effect are poorly understood. Additionally, abnormal activation of β-adrenergic signaling have been implicated in the development of pathological cardiac remodeling. However, the role of serine/threonine protein phosphatase 1 (PP1) in pathological cardiac remodeling under the influence of β-adrenergic signaling have been sparsely investigated...
2018: PloS One
Kumiko Nishimura, Masafumi Takeda, Jun K Yamashita, Ichiro Shiojima, Nagaoki Toyoda
AIMS: Type 3 iodothyronine deiodinase (D3), which converts thyroxine (T4 ) and 3,5,3'-triiodothyronine (T3 ) to 3,3',5'-triiodothyronine (rT3 ) and 3,3'-diiodothyronine (T2 ), respectively, inactivates thyroid hormones. We investigated the expression and regulation of D3 in human cardiomyocytes which were differentiated from human induced pluripotent stem cells (hiPSCs). MAIN METHODS: We characterized D3 activity using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis...
June 15, 2018: Life Sciences
Fan-Jing Meng, Jiang Cao, Chong Chen, Qing-Yun Wu, Xu-Guang Song, Wei Chen, Kai-Lin Xu, Wan-Chuan Zhuang
OBJECTIVE: To evaluate biological effects of OCT4A gene on K562 cells and explore the molecular mechanism of K562 cell apoptosis. METHODS: Two recombinant lentiviral vectors were constructed, which could stablely up- regulate and down- regulate OCT4A protein. Recombinant lentivirus was generated by co-transfection of three-plasmids and transfec-ted into K562 cells. The experiments were divided into 5 groups: normal, pLVX-OCT4A-ZsGreen1, pLVX vector control, PLB-OCT4A shRNA and non-specific shRNA groups...
April 2018: Zhongguo Shi Yan Xue Ye Xue za Zhi
Fernandes-Cerqueira Cátia, Quelhas-Santos Janete, Sampaio-Maia Benedita, Simões-Silva Liliana, Soares-Silva Isabel, Roncon-Albuquerque Roberto, Pestana Manuel
Puromycin aminonucleoside-induced nephrotic syndrome (PAN-NS) is characterized by cardiac remodeling and increased local inflammatory activity. Patients with NS and animal models of NS have vitamin D3 deficiency. The aim of the present study was to evaluate the influence of calcitriol on cardiac remodeling and local inflammatory state in PAN-NS rat model. Male Sprague-Dawley rats were injected with PAN or vehicle on day 0. PAN and control rats were divided into two subgroups for the administration of calcitriol (PAN-D and Ct-D groups) or the vehicle (PAN-V and Ct-V groups) during 21 days...
2018: BioMed Research International
Karem M Zekry, Norio Yamamoto, Katsuhiro Hayashi, Akihiko Takeuchi, Hiroyuki Tsuchiya
Introduction: Primary lymphoma of the bone (PLB) is uncommon clinical entity with the vast majority of the cases are non-Hodgkin's lymphoma. Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of PLB. Case Report: We report a case of 60-year-old female presented with pain in the right hip that persisted for 2 months. Plain X-ray, computed tomography, and magnetic resonance imaging of the right hip showed a lytic lesion involving the right acetabulum and adjoining iliac bone with extraosseous mass...
November 2017: Journal of Orthopaedic Case Reports
Andrea Roca-Dols, Marta Elena Losa-Iglesias, Rubén Sánchez-Gómez, Daniel López-López, Ricardo Becerro-de-Bengoa-Vallejo, César Calvo-Lobo
BACKGROUND: Peroneus longus and brevis (PLB) disorders are commonly in people with lateral ligamentous instability, ankle pain, lateral hindfoot pain and structures of the proximal compartment of the lower legs and their muscle activity is believed to be influenced by different footwear types. The proposal of this research is to evaluate the effects of five types of footwear with respect to the barefoot condition and analyze the activity patterns of PLB muscles in healthy subjects during the gait cycle...
June 2018: Journal of the Mechanical Behavior of Biomedical Materials
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