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measurement of ATP

Jean Pierre Oses, Alexandre Pastoris Müller, Nathan Ryzewski Strogulski, Julia D Moreira, Ana Elisa Böhmer, Gisele Hansel, Randhall Bruce Carteri, João Vicente Busnello, Afonso Kopczynski, Marcelo Salimen Rodolphi, Diogo Onofre Souza, Luis Valmor Portela
Generalized seizures trigger excessive neuronal firing that imposes large demands on the brain glucose/lactate availability and utilization, which synchronization requires an integral mitochondrial oxidative capability. We investigated whether a single convulsive crisis affects brain glucose/lactate availability and mitochondrial energy production. Adult male Wistar rats received a single injection of pentylentetrazol (PTZ, 60 mg/kg, i.p.) or saline. The cerebrospinal fluid (CSF) levels of glucose and lactate, mitochondrial respirometry, [14 C]-2-deoxy-D-glucose uptake, glycogen content and cell viability in hippocampus were measured...
March 11, 2019: Epilepsy Research
Lilin Ge, Bin Li, Houxi Xu, Wenyuan Pu, Hang Fai Kwok
A simple and feasible electrochemical immunosensing protocol with glucometer readout was designed for the detection of low-abundance disease-related biomarker (alpha-fetoprotein; AFP) on the basis of backfilling rolling cycle amplification (RCA) with invertase-DNA2 conjugates on the detection antibody. The assay consisted of the immunoreaction, RCA reaction, DNA2 -invertase hybridization and glucose measurement. Initially, a sandwiched immunoreaction was carried out between anti-AFP capture antibody-coated microplate between nanogold-labeled pAb2 detection antibody conjugated with DNA1 primer (DNA1 -AuNP-pAb2 ) in the presence of target ATP...
March 2, 2019: Biosensors & Bioelectronics
In-Sung Song, Yu Jeong Jeong, Seung Hun Jeong, Ji Eun Kim, Jin Han, Tae-Hyun Kim, Sung-Wuk Jang
BACKGROUND/AIMS: Breast cancer is a clinically and molecularly heterogeneous disease. Patients with triple-negative breast cancer (TNBC) have poorer outcomes than those with other breast cancer subtypes due to lack of effective molecular targets for therapy. The present study aimed to the identification of estrogen receptor (ER)β as a novel mitochondrial target in TNBC cells, together with underlying mechanisms. METHODS: Expression of ERβ in clinical breast samples were examined by qRT-PCR, immunohistochemistry and immunoblotting...
2019: Cellular Physiology and Biochemistry
Lingyu Li, Haomeng Yang, Lianwei Peng, Weibo Ren, Jirui Gong, Peng Liu, Xinhong Wu, Fang Huang
Sheepgrass [ Leymus chinensis (Trin.) Tzvel] is a valuable forage plant highly significant to the grassland productivity of Euro-Asia steppes. Growth of above-ground tissues of L. chinensis is the major component contributing to the grass yield. Although it is generally known that this species is sensitive to ecosystem disturbance and adverse environments, detailed information of how L. chinensis coping with various nutrient deficiency especially phosphate deprivation (-Pi) is still limited. Here, we investigated impact of Pi-deprivation on shoot growth and biomass accumulation as well as photosynthetic properties of L...
2019: Frontiers in Plant Science
Abhishek S Kashyap, Tamara Thelemann, Richard Klar, Sandra M Kallert, Julia Festag, Melanie Buchi, Lisa Hinterwimmer, Monika Schell, Sven Michel, Frank Jaschinski, Alfred Zippelius
BACKGROUND: Cancer cells are known to develop mechanisms to circumvent effective anti-tumor immunity. The two ectonucleotidases CD39 and CD73 are promising drug targets, as they act in concert to convert extracellular immune-stimulating ATP to adenosine. CD39 is expressed by different immune cell populations as well as cancer cells of different tumor types and supports the tumor in escaping immune recognition and destruction. Thus, increasing extracellular ATP and simultaneously reducing adenosine concentrations in the tumor can lead to effective anti-tumor immunity...
March 12, 2019: Journal for Immunotherapy of Cancer
Ladislav Valkovič, William T Clarke, Albrecht I Schmid, Betty Raman, Jane Ellis, Hugh Watkins, Matthew D Robson, Stefan Neubauer, Christopher T Rodgers
BACKGROUND: Cardiovascular phosphorus MR spectroscopy (31 P-CMRS) is a powerful tool for probing energetics in the human heart, through quantification of phosphocreatine (PCr) to adenosine triphosphate (ATP) ratio. In principle, 31 P-CMRS can also measure cardiac intracellular pH (pHi ) and the free energy of ATP hydrolysis (ΔGATP ). However, these require determination of the inorganic phosphate (Pi) signal frequency and amplitude that are currently not robustly accessible because blood signals often obscure the Pi resonance...
March 14, 2019: Journal of Cardiovascular Magnetic Resonance
Jeonghoon Han, Jun Chul Park, Hye-Min Kang, Eunjin Byeon, Deok-Seo Yoon, Min-Chul Lee, Alaa El-Din H Sayed, Un-Ki Hwang, Jae-Seong Lee
To examine the adverse effects of the benzo[α]pyrene (B[α]P), the monogonont rotifer Brachionus rotundiformis was exposed to various concentration of B[α]P (0 [control], 1, 10, and 100 μg/L) and measured life cycle parameters (e.g., mortality, fecundity [cumulated number of offspring], and lifespan), reactive oxygen species (ROS), antioxidant enzymatic activity of glutathione S-transferase (GST). In addition, defense-related transcripts (e.g., glutathione S-transferases [GSTs], ATP binding cassette [ABCs] transporters) and Western blot analysis of mitogen-activated protein kinase (MAPK) signaling pathway were investigated in B[α]P-exposed rotifer...
March 6, 2019: Aquatic Toxicology
Kazuhiro Nakagawa, Takuya Kishimoto
The need for technologies to monitor cell health is increasing with advancements in the field of cell therapy and regenerative medicine. In this study, we demonstrated unlabeled optical metabolic imaging of cultured living cells. This imaging technique is based on motion vector analysis with a block-matching algorithm to compare sequential time-lapse images. Motion vector analysis evaluates the movement of intracellular granules observed with a phase-contrast microscope. We demonstrated that the motion speed of intracellular movement reflects adenosine triphosphate (ATP)-dependent intracellular trafficking in cells...
March 2019: BioTechniques
Cornelia Wiegand, Martin Abel, Uta-Christina Hipler, Peter Elsner
Non-adhering dressings are commonly used during granulation, tissue formation, and re-epithelialization. Elucidating cytotoxic effects and influence on proliferation/migration capacity of cells like fibroblasts is of interest. Dressings' effects were investigated by comprehensive in vitro approach: (1) MTT assay measuring cell viability after direct contact, (2) ATP assay determining effects on cell proliferation, and (3) scratch wound assay featuring an in vitro wound healing model. One cotton-based dressing with vaseline (vas) was included in the study and four polyester dressings containing vas and technology-lipido-colloid matrix (TLC), carboxymethylcellulose (CMC), hydrocolloid (HC), or glycerin (gly) as additives...
March 13, 2019: Scientific Reports
Kun Yin, Guihua Zhao, Chao Xu, Xiao Qiu, Biwei Wen, Hui Sun, Gongzhen Liu, Ye Liu, Qingsong Zhao, Qingkuan Wei, Bingcheng Huang, Ge Yan, Jianping Cao
BACKGROUND: Rhoptry protein 18 (ROP18) is a key virulence factor of Toxoplasma gondii. The host's immune responses mediated by immune-related GTPases (IRGs) could be blocked by ROP18's kinase activity. ROP18 also interacts with various substrates, such as activating transcription factor 6 beta (ATF6β) and affects multiple physiological functions within host cells, thereby inducing intense virulence. In this study, competitive inhibitors targeted to ROP18 were subjected to virtual screening based on the principle of structure-based drug design (SBDD)...
March 13, 2019: Parasites & Vectors
Manuel Belli, Ling Zhang, Xiaowei Liu, Annemarie Donjacour, Elena Ruggeri, Maria Grazia Palmerini, Stefania Annarita Nottola, Guido Macchiarelli, Paolo Rinaudo
STUDY QUESTION: Does the oxygen concentration in the culture medium [either physiologic (5%) or atmospheric (20%)] affect mitochondrial ultrastructure and function in preimplantation mouse embryos generated by IVF? SUMMARY ANSWER: Embryos cultured in 20% oxygen show increased mitochondrial abnormalities compared to embryos cultured in 5% oxygen. WHAT IS KNOWN ALREADY: ART are widely used and have resulted in the birth of more than 8 million children...
March 13, 2019: Human Reproduction
Shufen Han, Wei Zhang, Ru Zhang, Jun Jiao, Chunling Fu, Xing Tong, Weiguo Zhang, Liqiang Qin
Background: Dietary intake of cereal fiber has been reported to benefit lipid metabolism through multiple mechanisms. The present study aimed to discover the potential mechanisms by which cereal fiber could modify the intestinal cholesterol metabolism. Design: Male C57BL/6 mice were fed a reference chow (RC) diet; high-fat, high-cholesterol (HFC) diet; HFC plus oat fiber diet; or HFC plus wheat bran fiber diet for 24 weeks. Serum lipids were measured by enzymatic methods...
2019: Food & Nutrition Research
Nicholas Dale, Faming Tian, Ravjit Sagoo, Norman Phillips, Chris Imray, Christine Roffe
Stroke is a leading cause of death and disability. Here, we examine whether point-of-care measurement of the purines, adenosine, inosine and hypoxanthine, which are downstream metabolites of ATP, has potential to assist the diagnosis of stroke. In a prospective observational study, patients who were suspected of having had a stroke, within 4.5 h of symptom onset and still displaying focal neurological symptoms at admission, were recruited. Clinical research staff in the Emergency Departments of two hospitals used a prototype biosensor array, SMARTCap, to measure the purines in the venous blood of stroke patients and healthy controls...
March 12, 2019: Purinergic Signalling
Liam Curry, Hani Almukhtar, Jala Alahmed, Richard Roberts, Paul A Smith
Plasma membrane ion channels and mitochondrial electron transport complexes (mETC) are recognised "off-targets" for certain drugs. Simvastatin is one such drug, a lipophilic statin used to treat hypercholesterolaemia, but which is also associated with adverse effects like myopathy and increased risk of glucose intolerance. Such myopathy is thought to arise through adverse actions of simvastatin on skeletal muscle mETC and mitochondrial respiration. In this study we investigated whether the glucose intolerance associated with simvastatin is also mediated via adverse effects on mETC in pancreatic beta-cells since mitochondrial respiration underlies insulin secretion from these cells, an effect in part mediated by promotion of Ca2+ influx via opening of voltage-gated Ca2+ channels (VGCCs)...
March 11, 2019: Toxicological Sciences: An Official Journal of the Society of Toxicology
Nazia Naz S Khan, Karen Kelly-Blake, Zhehui Luo, Adesuwa Olomu
Purpose: The purpose of this study was to determine the difference in the rate of statin prescribing based on the Adult Treatment Panel (ATP) III and 2013 American College of Cardiology (ACC)/American Heart Association cholesterol guidelines across sex in Federally Qualified Health Centers (FQHCs), and to determine the proportion of patients on recommended statin dosage based on the 2013 cholesterol guideline. Methods: The Office Guidelines Applied to Practice (Office-GAP) study is a quasi-experimental, 2 FQHCs center study that enrolled patients with coronary heart disease and diabetes mellitus (DM)...
January 2019: Health Services Research and Managerial Epidemiology
Caroline Di Tommaso, Liz Taylor-Edmonds, Susan A Andrews, Robert C Andrews
Biofiltration has been shown to be effective for disinfection by-product (DBP) precursor control, however few studies have considered its role in the potential formation of DBPs. Biofilm is composed of heterogeneous bacteria as well as extracellular polymeric substances (EPS). The objective of this study was to determine the contribution of biofilm-related materials such as EPS to form nitrogen-containing DBPs upon chloramination, and to determine the influence of cyclical (scheduled on-off) biofilter operation on DBP precursor removal...
February 26, 2019: Water Research
Ievgen Mazurenko, Nikos S Hatzakis, Lars J C Jeuken
Proton-pumping enzymes of electron transfer chains couple redox reactions to proton translocation across the membrane, creating a proton-motive force used for ATP production. The amphiphilic nature of membrane proteins requires particular attention to their handling, and reconstitution into the natural lipid environment is indispensable when studying membrane transport processes like proton translocation. Here, we detail a method that has been used for the investigation of the proton-pumping mechanism of membrane redox enzymes, taking cytochrome bo3 from Escherichia coli as an example...
February 21, 2019: Journal of Visualized Experiments: JoVE
Morgan B Vorwald, Lisa C Letzkus
The aim of this project is to determine the amount of organic matter burden focused on exercise-tracking devices worn by heath care professionals in the clinical setting. It was completed by using adenosine triphosphate surveillance, which measures the residual organic matter that remains on a device. Because all but 1 of the devices sampled were considered contaminated, health care professionals should be aware of the potential risk associated with wearing exercise-tracking devices in the clinical setting.
March 5, 2019: American Journal of Infection Control
Bernard P Binks, Hui Shi
An aqueous two-phase system (ATPS) is of great value in low calorie foods or oil-free cosmetics and pharmaceuticals. In contrast to the recent work on polymer/polymer ATPSs, a simple polymer/salt ATPS (polyethylene glycol/Na2 SO4 ) was chosen to study water-in-water (w/w) emulsions stabilized by solid particles. The binodal curve and the tie lines were first determined for the mixture at room temperature. Above the binodal curve, two water-based phases coexist; the upper phase is rich in polymer, whereas the lower phase is rich in salt...
March 8, 2019: Langmuir: the ACS Journal of Surfaces and Colloids
Li You, Ying-Ying Pan, Meng-Yao An, Wen-Hua Chen, Ying Zhang, Yan-Na Wu, Yan Li, Kai Sun, Yong-Qiang Yin, Jian-Shi Lou
BACKGROUND Cardiac remote ischemic conditioning (RIC) is a noninvasive cardioprotective method in ischemia-reperfusion injury and acute myocardial infarction (AMI). The aims of this study were to investigate the effects of RIC in a rat model of AMI. MATERIAL AND METHODS Adult male Sprague-Dawley rats included the AMI group that underwent ligation of the left anterior descending (LAD) coronary artery (n=24), the RIC group that consisted the AMI rat model treated with RIC once daily in the left hind limb until days 1, 7 and 14 (n=24), and the sham group (n=24)...
March 8, 2019: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
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