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Downstream processing

Arun Kumar C Huded, Pavankumar Jingade, Manoj Kumar Mishra
Isolation of high-quality RNA from coffee is challenging because of high level of polysaccharides, polyphenols and other secondary metabolites. In the present study, a rapid and efficient RNA extraction protocol from different tissues of coffee was optimized. Sufficiently high quality and quantity (225.6-454.8 µg/g) of RNA was obtained by using the optimized protocol. The presence of two distinct bands of 28S rRNA and 18S rRNA in agarose gel proved the intactness of the RNA samples. The average spectrophotometric values of the isolated RNA ranged from 1...
March 2018: 3 Biotech
Jun Liu, Lei Huang, Pengxiao Su, Tao Song, Wentao Zhang, Jinzhu Fan, Yang Liu
A number of studies have attempted to elucidate the association between mircoRNAs (miRNAs/miRs) and cancer-associated processes. The aim of the present study was to determine how miR-499a-5p intervenes in human osteosarcoma cell proliferation and differentiation. The cancerous tissues and adjacent non-cancerous tissues of 62 patients with osteosarcoma (OS) were collected. miRNA microarray analysis revealed that 29 miRNAs were upregulated while 26 were downregulated, among which miR-499a-5p expression was the most decreased...
April 2018: Oncology Letters
D Podbevsek, D Colombet, G Ledoux, F Ayela
We have performed hydrodynamic cavitation experiments with an aqueous luminol solution as the working fluid. Light emission, together with the high frequency noise which characterizes cavitation, was emitted by the two-phase flow, whereas no light emission from luminol was recorded in the single phase liquid flow. Light emission occurs downstream transparent microdiaphragms. The maximum level of the recorded signal was around 180 photons per second with flow rates of 380 µl/s, that corresponds to a real order of magnitude of the chemiluminescence of 75,000 photons per second...
May 2018: Ultrasonics Sonochemistry
Cunzhen Shi, Xiangqun Li, Xiaogan Wang, Ning Ding, Lingyan Ping, Yunfei Shi, Lan Mi, Yumei Lai, Yuqin Song, Jun Zhu
BACKGROUND: Mantle cell lymphoma (MCL) is an incurable B cell-derived malignant tumor with a median overall survival of 4-5 years. Mer tyrosine kinase (MerTK) has been reported to be aberrantly expressed in leukemia, melanoma, and gastric cancer, and plays a pivotal role in the process of oncogenesis. This study assessed the role of MerTK in MCL for the first time. METHODS: Immunohistochemistry and western blot were performed to figure out expression of MerTK in MCL...
March 20, 2018: Journal of Hematology & Oncology
Andrea Luca Tasca, Monica Puccini, Ashleigh Fletcher
The herbicide terbuthylazine (TBA) has displaced atrazine in most of EU countries, becoming one of the most regularly used pesticides and, therefore, frequently detected in natural waters. The affinity of TBA for soil organic matter suggests prolonged contamination; degradation leads to the release of the metabolite desethylterbuthylazine (DET), which has higher water solubility and binds more weakly to organic matter compared to the parent compound, resulting in higher associated risk for contamination of groundwater resources...
March 14, 2018: Chemosphere
Fatima Cabral, Colton M Miller, Katrina M Kudrna, Blake E Hass, Jocelyn G Daubendiek, Brianna M Kellar, Edward N Harris
This protocol demonstrates a method for obtaining high yield and viability for mouse hepatocytes and sinusoidal endothelial cells (SECs) suitable for culturing or for obtaining cell lysates. In this protocol, the portal vein is used as the site for catheterization, rather than the vena cava, as this limits contamination of other possible cell types in the final liver preparation. No special instrumentation is required throughout the procedure. A water bath is used as a source of heat to maintain the temperature of all the buffers and solutions...
February 12, 2018: Journal of Visualized Experiments: JoVE
Justin Weinberg, Shaojie Zhang, Allison Kirkby, Enosh Shachar, Giorgio Carta, Todd Przybycien
We have proposed chemical modification of Protein A (ProA) chromatography ligands with polyethylene glycol (PEGylation) as a strategy to increase the resin selectivity and robustness by providing the ligand with a steric repulsion barrier against non-specific binding. Here, we report on robustness and selectivity benefits for Repligen CaptivA PriMAB resin with ligands modified with 5.2 kDa and 21.5 kDa PEG chains, respectively. PEGylation of ProA ligands allowed the resin to retain a higher percentage of static binding capacity relative to the unmodified resin upon digestion with chymotrypsin, a representative serine protease...
February 24, 2018: Journal of Chromatography. A
Aritri Sanyal, Runa Antony, Gautami Samui, Meloth Thamban
Cryoconite holes (cylindrical melt-holes on the glacier surface) are important hydrological and biological systems within glacial environments that support diverse microbial communities and biogeochemical processes. This study describes retrievable heterotrophic microbes in cryoconite hole water from three geographically distinct sites in Antarctica, and a Himalayan glacier, along with their potential to degrade organic compounds found in these environments. Microcosm experiments (22 days) show that 13-60% of the dissolved organic carbon in the water within cryoconite holes is bio-available to resident microbes...
March 2018: Microbiological Research
Christopher S Jones, Sea-Won Kim, Thomas F Wilton, Keith E Schilling, Caroline A Davis
Real-time, continuous, in situ water quality sensors were deployed on a fourth-order Iowa (U.S.) stream draining an agricultural watershed to evaluate key in-stream processes affecting concentrations of nitrate during a 24-day late summer (Aug-Sep) period. Overall, nitrate-nitrogen (NO3 -N) concentrations declined 0.11 mg L-1  km-1 , or about 1.9% km-1 and 35% in total across 18 km. We also calculated stream metabolic rates using in situ dissolved oxygen data and determined stream biotic N demand to be 108-117 mg m-2  day-1 ...
March 17, 2018: Environmental Monitoring and Assessment
Yi-Mei J Lin, I-Lun Hsin, H Sunny Sun, Shankung Lin, Yen-Ling Lai, Hsuan-Ying Chen, Ting-Yu Chen, Ya-Ping Chen, Yi-Ting Shen, Hung-Ming Wu
POU-homeodomain transcription factor POU3F2 is a critical transcription factor that participates in neuronal differentiation. However, little is known about its downstream mediators. Here genome-wide analyses of a human neuronal differentiation cell model, NT2D1, suggested neurotrophin-3 (NTF3), a key mediator of neuronal development during the early neurogenic period, as a putative regulatory target of POU3F2. Western blot, cDNA microarray, and real-time quantitative PCR analyses showed that POU3F2 and NTF3 were upregulated during neuronal differentiation...
March 16, 2018: Molecular Neurobiology
Zachary S Lorsch, Yong-Hwee Eddie Loh, Immanuel Purushothaman, Deena M Walker, Eric M Parise, Marine Salery, Michael E Cahill, Georgia E Hodes, Madeline L Pfau, Hope Kronman, Peter J Hamilton, Orna Issler, Benoit Labonté, Ann E Symonds, Matthew Zucker, Tie Yuan Zhang, Michael J Meaney, Scott J Russo, Li Shen, Rosemary C Bagot, Eric J Nestler
Most people exposed to stress do not develop depression. Animal models have shown that stress resilience is an active state that requires broad transcriptional adaptations, but how this homeostatic process is regulated remains poorly understood. In this study, we analyze upstream regulators of genes differentially expressed after chronic social defeat stress. We identify estrogen receptor α (ERα) as the top regulator of pro-resilient transcriptional changes in the nucleus accumbens (NAc), a key brain reward region implicated in depression...
March 16, 2018: Nature Communications
Beate Hoffmann, Frank Löhr, Aisha Laguerre, Frank Bernhard, Volker Dötsch
Preparation of a protein sample for liquid-state nuclear magnetic resonance (NMR) spectroscopy analysis requires optimization of many parameters. This review describes labeling strategies for obtaining assignments of protein resonances. Particular emphasis is placed on the advantages of cell-free protein production, which enables exclusive labeling of the protein of interest, thereby simplifying downstream processing steps and increasing the availability of different labeling strategies for a target protein...
April 2018: Progress in Nuclear Magnetic Resonance Spectroscopy
Veronika Dill, Bernd Hoffmann, Aline Zimmer, Martin Beer, Michael Eschbaumer
BACKGROUND: Suspension culture of BHK cells allows large-scale virus propagation and cost-efficient vaccine production, while the shift to animal-component-free cell culture media without serum is beneficial for the quality and downstream processing of the product. Foot-and-mouth disease virus is still endemic in many parts of the world and high-quality vaccines are essential for the eradication of this highly contagious and economically devastating disease. METHODS: Changes to the viral genome sequence during passaging in an adherent and a suspension cell culture system were compared and the impact of amino acid substitutions on receptor tropism, antigenicity and particle stability was examined...
March 16, 2018: Virology Journal
Sandeep Choubey
Regulation of transcription is a vital process in cells, but mechanistic details of this regulation still remain elusive. The dominant approach to unravel the dynamics of transcriptional regulation is to first develop mathematical models of transcription and then experimentally test the predictions these models make for the distribution of mRNA and protein molecules at the individual cell level. However, these measurements are affected by a multitude of downstream processes which make it difficult to interpret the measurements...
February 2018: Physical Review. E
Martin Kornecki, Jochen Strube
Productivity improvements of mammalian cell culture in the production of recombinant proteins have been made by optimizing cell lines, media, and process operation. This led to enhanced titers and process robustness without increasing the cost of the upstream processing (USP); however, a downstream bottleneck remains. In terms of process control improvement, the process analytical technology (PAT) initiative, initiated by the American Food and Drug Administration (FDA), aims to measure, analyze, monitor, and ultimately control all important attributes of a bioprocess...
March 16, 2018: Bioengineering
Amjad M Husaini, Aafreen Sakina, Souliha R Cambay
Fusarium oxysporum, a ubiquitous soil-borne pathogen causes devastating vascular wilt in more than 100 plant species and ranks fifth among top ten fungal plant pathogens. It has emerged as a human pathogen too, causing infections in immune-compromised patients. It is, therefore, important to gain insight into the molecular processes involved in the pathogenesis of this trans-kingdom pathogen. A complex network comprising of interconnected and over lapping signal pathways; mitogen-activated protein kinase (MAPK) signaling pathways, Ras proteins, G-protein signaling components and their downstream pathways, components of the velvet (LaeA/VeA/VelB) complex and cAMP pathways, is involved in perceiving the host...
March 16, 2018: Molecular Plant-microbe Interactions: MPMI
Deepali Jain, Sinchita Roy-Chowdhuri
CONTEXT: - There has been a paradigm shift in the understanding of molecular pathogenesis of lung cancer. A number of oncogenic drivers have been identified in non-small cell lung carcinoma, such as the epidermal growth factor receptor ( EGFR) mutation and anaplastic lymphoma kinase ( ALK) gene rearrangement. Because of the clinical presentation at an advanced stage of disease in non-small cell lung carcinoma patients, the use of minimally invasive techniques is preferred to obtain a tumor sample for diagnosis...
March 16, 2018: Archives of Pathology & Laboratory Medicine
Adam S Lowet, Chaz Firestone, Brian J Scholl
An intrinsic part of seeing objects is seeing how similar or different they are relative to one another. This experience requires that objects be mentally represented in a common format over which such comparisons can be carried out. What is that representational format? Objects could be compared in terms of their superficial features (e.g., degree of pixel-by-pixel overlap), but a more intriguing possibility is that they are compared on the basis of a deeper structure. One especially promising candidate that has enjoyed success in the computer vision literature is the shape skeleton-a geometric transformation that represents objects according to their inferred underlying organization...
March 15, 2018: Attention, Perception & Psychophysics
H Zhao, J Wang, D Fang, O Lee, R T Chatterton, V Stearns, S A Khan, S E Bulun
Obesity is associated with increased risk of breast cancer in postmenopausal but not in premenopausal women. Many factors may be responsible for this difference. The aim of this study was to determine the mechanisms by which the genes related to the AMPK pathway, inflammation, and estrogen actions are affected by adiposity in breast tissue with the objective of identifying differences that may explain the different breast cancer risk in premenopausal and postmenopausal women. Random fine needle aspirates (rFNAs) of breast tissue were collected from 57 premenopausal and 55 postmenopausal women and were classified as normal weight, overweight, or obese...
March 15, 2018: Hormones & Cancer
Ana P F Santos, Bianca M Souza, Tânia F C V Silva, Rodrigo P Cavalcante, Silvio C Oliveira, Amílcar Machulek, Rui A R Boaventura, Vítor J P Vilar
The mineralization of bio-recalcitrant humic acids (HAs) by a solar photo-Fenton (SPF) process was investigated in aqueous system, in order to understand its abatement in real high-HA content matrices, such as sanitary landfill leachates. SPF reactions were performed in tubular photoreactors with CPCs at lab-scale (simulated solar light) and pilot-scale (natural sunlight). Considering the experimental conditions selected for this work, the formation of insoluble HA-Fe3+ complexes was observed. Thus, to avoid HA precipitation, oxalic acid (Ox) was added, since Fe3+ -Ox complexes present a higher stability constant...
March 15, 2018: Environmental Science and Pollution Research International
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