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Gabriel Lima-Oliveira, Dorothy M Adcock, Gian Luca Salvagno, Emmanuel J Favaloro, Giuseppe Lippi
OBJECTIVE: Thus study was aimed to investigate whether the mixing technique could influence the results of routine and specialized clotting tests on post-thawed specimens. METHODS: The sample population consisted of 13 healthy volunteers. Venous blood was collected by evacuated system into three 3.5mL tubes containing 0.109mmol/L buffered sodium citrate. The three blood tubes of each subject were pooled immediately after collection inside a Falcon 15mL tube, then mixed by 6 gentle end-over-end inversions, and centrifuged at 1500g for 15min...
October 18, 2016: Clinical Biochemistry
Huiduo Guo, Yabin Gong, Bin He, Ruqian Zhao
Energy produced by mitochondria via oxidative phosphorylation (OXPHOS) is essential for mammalian sperm motility. Mammalian mitochondrial DNA (mtDNA)-encoded proteins are subunits of the OXPHOS system. Paradoxically, there are less mitochondrial and mtDNA contents in motile sperm than less motile sperm. Here, mature boar sperm was used as a model to investigate the relationships between mtDNA content, mitochondrial activity, and sperm motility. Motile and less motile sperm were separated by centrifugation on a discontinuous percoll density gradient...
September 13, 2016: Theriogenology
Zhongling Liu, Wei Yu, Hanqi Zhang, Fanbin Gu, Xiangqun Jin
Salting-out homogenous extraction followed by ionic liquid/ionic liquid dispersive liquid-liquid micro-extraction system was developed and applied to the extraction of sulfonamides in blood. High-performance liquid chromatography was applied to the determination of the analytes. The blood sample was centrifuged to obtain the serum. After the proteins in the serum were removed in the presence of acetonitrile, ionic liquid 1-butyl-3-methylimidazolium tetrafluoroborate, dipotassium hydrogen phosphate, ionic liquid 1-Hexyl-3-methylimidazolium hexafluorophosphate were added into the resulting solution...
December 1, 2016: Talanta
Anas Alshishani, Ahmad Makahleh, Hui Fang Yap, Elbaleeq Adam Gubartallah, Salizawati Muhamad Salhimi, Bahruddin Saad
A new sample preparation method, ion-pair vortex assisted liquid-liquid microextraction (VALLME-BE), for the determination of a highly polar anti-diabetic drug (metformin) in plasma sample was developed. The VALLME-BE was performed by diluting the plasma in borate buffer and extracted to 150µL 1-octanol containing 0.2M di-(2-ethylhexyl)phosphoric acid as intermediate phase. The drug was next back-extracted into 20µL of 0.075M HCl solution. The effects of pH, ion-pair concentration, type of organic solvent, volume of extraction phases, ionic strength, vortexing and centrifugation times on the extraction efficiency were investigated...
December 1, 2016: Talanta
Per J R Sjöberg, Pernilla Thelin, Emil Rydin
A method for isomeric separation of inositol phosphates (InsPn) in environmental samples originating from different sources such as soil, manure/compost, and aquatic sediments has been developed. The method includes a single NaOH-EDTA extraction step, centrifugation and direct injection of a particle free solution into an ion chromatographic column. Isomeric liquid chromatographic separation was achieved with an ammonium carbonate gradient compatible with electrospray ionization tandem mass spectrometric detection (LC-ESI-MS/MS)...
December 1, 2016: Talanta
Magdalena Krawczyk, Ewa Stanisz
The combination of ultrasound-assisted dispersive micro solid-phase extraction (USA DMSPE), with the use of TiO2 nanoparticles (NPs) as adsorbent, with cold vapor atomic absorption spectrometry (CV AAS) is proposed for preconcentration and determination of mercury species (Hgtotal, Hg(2+) and CH3Hg(+)) in biological, geological and water samples. The experimental parameters including the amount of TiO2 NPs, pH of sample solution, ultrasonication and centrifugation time, TiO2 slurry solution preparation before injection to CV AAS were investigated...
December 1, 2016: Talanta
Man Wang, Ping Ma, Xiaonan Xi, Lei Liu, Yabin Wen, Kangning Liu, Liang Sun, Yaxin Lu, Zheng Yin
The bioanalysis and especially the sample preparation of nucleoside drugs in complex media, such as human plasma, has been challenging due to the high polarity and high solubility of these drugs in water. Online solid phase extraction (SPE) offers significant advantages, such as automation and timesaving. Thus, several types of SPE columns have been developed for compounds with different polarities. In this study, SPE was applied to overcome the issue of sample pretreatment of nucleoside drugs in human plasma, with the final aim of establishing a robust analytical platform for drugs with similar structures...
December 1, 2016: Talanta
Na Wei, Xian-En Zhao, Shuyun Zhu, Yongrui He, Longfang Zheng, Guang Chen, Jinmao You, Shu Liu, Zhiqiang Liu
This paper, for the first time, reported a simple, rapid, sensitive and environmental friendly ultrasonic-assisted in situ derivatization-dispersive liquid-liquid microextraction (in situ UAD-DLLME) method followed by ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) for the simultaneous determination of dopamine (DA), serotonin (5-HT) and their biosynthesis precursors and metabolites in rat brain microdialysates. In this work, a commercial reagent, Lissamine rhodamine B sulfonylchloride (LRSC), was proposed as a derivatization reagent...
December 1, 2016: Talanta
Simon Felz, Salah Al-Zuhairy, Olav Andreas Aarstad, Mark C M van Loosdrecht, Yue Mei Lin
To evaluate and develop methodologies for the extraction of gel-forming extracellular polymeric substances (EPS), EPS from aerobic granular sludge (AGS) was extracted using six different methods (centrifugation, sonication, ethylenediaminetetraacetic acid (EDTA), formamide with sodium hydroxide (NaOH), formaldehyde with NaOH and sodium carbonate (Na2CO3) with heat and constant mixing). AGS was collected from a pilot wastewater treatment reactor. The ionic gel-forming property of the extracted EPS of the six different extraction methods was tested with calcium ions (Ca(2+))...
September 26, 2016: Journal of Visualized Experiments: JoVE
Brandon L Thompson, Christopher Birch, Daniel A Nelson, Jingyi Li, Jacquelyn A DuVall, Delphine Le Roux, An-Chi Tsuei, Daniel L Mills, Brian E Root, James P Landers
Current conventional methods utilized for forensic DNA analysis are time consuming and labor-intensive requiring large and expensive equipment and instrumentation. While more portable Rapid DNA systems have been developed, introducing them to a working laboratory still necessitates a high cost of initiation followed by the recurrent cost of the devices. This has highlighted the need for an inexpensive, rapid and portable DNA analysis tool for human identification in a forensic setting. In order for an integrated DNA analysis system such as this to be realized, device operations must always be concluded by a rapid separation of short-tandem repeat (STR) DNA fragments...
October 21, 2016: Lab on a Chip
Ilhem Guesmi-Mzoughi, Antonio Archidona-Yuste, Carolina Cantalapiedra-Navarrete, Hajer Regaieg, Najet Horrigue-Raouani, Juan E Palomares-Rius, Pablo Castillo
Spiral nematode species of the genus Rotylenchus have been reported on olive (Olea europaea L.) in several Mediterranean countries (Castillo et al., 2010; Ali et al., 2014). Nematological surveys for plant-parasitic nematodes on olive trees were carried out in Tunisia between 2013 and 2014, and two nematode species of Rotylenchus were collected from the rhizosphere of olive cv. Chemlali in several localities of Tunisia (Tables 1,2 [Table: see text] [Table: see text] ). Twenty-two soil samples of 3 to 4 kg were collected with a shovel from the upper 50 cm of soil from arbitrarily chosen olive trees...
September 2016: Journal of Nematology
Cherie Walker, Peter Sinclair, Kenneth Graham, Stephen Cobley
Inertial Measurement Units (IMUs) may offer an ecologically valid, reliable, and practical method for biomechanical performance analysis. With such potential in mind, Part 1 of this study examined the accuracy of IMUs gyroscopes with an optical system (Cortex 3.3). A calibration formula standardised the IMUs angular velocity output with the optical system. The percentage differences between the two measures = 0.5% (p < 0.05), suggest IMU's efficacy for application. In Part 2, the aim was to examine and understand how dive flight angular velocity time series plots change and increase according to dive degree of difficulty...
October 20, 2016: Sports Biomechanics
Ahmet Bekdemir, Francesco Stellacci
Nanomedicine requires in-depth knowledge of nanoparticle-protein interactions. These interactions are studied with methods limited to large or fluorescently labelled nanoparticles as they rely on scattering or fluorescence-correlation signals. Here, we have developed a method based on analytical ultracentrifugation (AUC) as an absorbance-based, label-free tool to determine dissociation constants (KD), stoichiometry (Nmax), and Hill coefficient (n), for the association of bovine serum albumin (BSA) with gold nanoparticles...
October 20, 2016: Nature Communications
Takeshi Harada, Hideki Yamamoto, Shosei Kishida, Michiko Kishida, Chihiro Awada, Toshifumi Takao, Akira Kikuchi
Wnt5b, a member of the same family of proteins as Wnt5a of which overexpression is associated with cancer aggressiveness, is suggested to be involved in cancer progression, however details remain unclarified. We analyzed biochemical properties of purified Wnt5b and the mode of secretion of Wnt5b by cancer cells. Wnt5b was glycosylated at three asparagine residues and lipidated at one serine residue, and these post-translational modifications of Wnt5b are essential for secretion. Purified Wnt5b showed Dvl2 phosphorylation and Rac activation abilities to a similar extent as Wnt5a...
October 20, 2016: Cancer Science
Antonella Caivano, Francesco La Rocca, Vittorio Simeon, Marco Girasole, Simone Dinarelli, Ilaria Laurenzana, Angelo De Stradis, Luciana De Luca, Stefania Trino, Antonio Traficante, Giovanni D'Arena, Giovanna Mansueto, Oreste Villani, Giuseppe Pietrantuono, Luca Laurenti, Luigi Del Vecchio, Pellegrino Musto
PURPOSE: The use of extracellular vesicles (EVs) from body fluids as "liquid biopsies" is emerging as a promising approach for the diagnosis, prognosis and therapeutic monitoring of cancer patients. MicroRNA-155 (miR155), a non-coding transcript of the B-cell integration cluster (BIC) gene, has been reported to play a critical role in the pathogenesis of several types of hematologic malignancies (HMs) in which high miR155 levels have been found. At yet, however, the EV miR155 level and its putative clinical relevance in sera of HM patients have not been reported...
October 19, 2016: Cellular Oncology (Dordrecht)
Chien-Cheng Chen, Christopher J Bockisch, Dominik Straumann, Melody Ying-Yu Huang
Spontaneous eye movements of zebrafish larvae in the dark consist of centrifugal saccades that move the eyes from a central to an eccentric position and postsaccadic centripetal drifts. In a previous study, we showed that the fitted single-exponential time constants of the postsaccadic drifts are longer in the temporal-to-nasal (T->N) direction than in the nasal-to-temporal (N->T) direction. In the present study, we further report that saccadic peak velocities are higher and saccadic amplitudes are larger in the N->T direction than in the T->N direction...
2016: Frontiers in Systems Neuroscience
Alberto de Castro Pochini, Eliane Antonioli, Daniella Zanetti Bucci, Luiz Roberto Sardinha, Carlos Vicente Andreoli, Mario Ferretti, Benno Ejnisman, Anna Carla Goldberg, Moisés Cohen
Objective: To evaluate growth factors and cytokines in samples of platelet-rich plasma obtained by three different centrifugation methods. Methods: Peripheral blood of six individuals with no hematological diseases, aged 18 to 68 years, was drawn to obtain platelet-rich plasma, using the open method and commercial columns by Medtronic and Biomet. The products obtained with the different types of centrifugation were submitted to laboratory analysis, including pro-inflammatory cytokines and chemokines by flow cytometry assays, the concentration of fibroblast growth factors-2 (FGF-2) and transforming growth factor-beta1 (TGF-β1)...
July 2016: Einstein
Sérgio Monteiro de Almeida, Indianara Rotta, Arnaldo José de Conto, Dario Antonelli, Carlos Dabdoub Roda, Edna Yoshiko Yamada, Gisele M B Singer
Objective: To define how to best handle cerebrospinal fluid (CSF) specimens to obtain the highest positivity rate for the diagnosis of malignancy, comparing two different methods of cell concentration, sedimentation and cytocentrifugation. Methods: A retrospective analysis of 411 CSF reports. Results: This is a descriptive comparative study. The positive identification of malignant CSF cells was higher using the centrifuge than that using the Suta chamber (27...
October 2016: Arquivos de Neuro-psiquiatria
W Tipkantha, P Thuwanut, B Siriaroonrat, P Comizzoli, K Chatdarong
Clouded leopards (Neofelis nebulosa) produced high proportion of abnormal spermatozoa (mainly tail defects) that can limit sperm movement and conception. The study aimed to better identify the origin of those defects using a demembranation approach. Ejaculates (1-2 ejaculations/male; n = 9) were allocated to simple washing (control; resulting in 11.7% ± 1.9% coiled tails) and processed through colloid centrifugation to reduce the number of sperm with tail defects (treatment, resulting in 5.9% ± 0.9% coiled tails)...
October 18, 2016: Reproduction in Domestic Animals, Zuchthygiene
Alexandra Condé-Green, Vasanth S Kotamarti, Lauren S Sherman, Jonathan D Keith, Edward S Lee, Mark S Granick, Pranela Rameshwar
: Standard isolation of adipose stromal vascular fraction (SVF) requires the use of collagenase and is considered more than "minimally manipulated" by current good manufacturing practice requirements. Alternatively, nonenzymatic isolation methods have surfaced using physical forces to separate cells from the adipose matrix. The purpose of this study was to review the literature on the use of mechanical isolation protocols and compare the results. The implication for use as a standard procedure in practice is discussed...
September 2016: Plastic and Reconstructive Surgery. Global Open
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