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https://www.readbyqxmd.com/read/28093272/a-method-for-obtaining-simian-immunodeficiency-virus-rna-sequences-from-laser-capture-microdissected-and-immune-captured-cd68-and-cd163-macrophages-from-frozen-tissue-sections-of-bone-marrow-and-brain
#1
Jaclyn Mallard, Emily Papazian, Caroline Soulas, David J Nolan, Marco Salemi, Kenneth C Williams
Laser capture microdissection (LCM) is used to extract cells or tissue regions for analysis of RNA, DNA or protein. Several methods of LCM are established for different applications, but a protocol for consistently obtaining lentiviral RNA from LCM captured immune cell populations is not described. Obtaining optimal viral RNA for analysis of viral genes from immune-captured cells using immunohistochemistry (IHC) and LCM is challenging. IHC protocols have long antibody incubation times that increase risk of RNA degradation...
January 13, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28066594/cough-frequency-monitors-can-they-discriminate-patient-from-environmental-coughs
#2
Stefan T Kulnik, Natalie M Williams, Lalit Kalra, John Moxham, Surinder S Birring
BACKGROUND: Objective cough frequency measurements are increasingly applied in clinical research. Technological advances enable automated detection and counting of cough events from sound recordings of many hours' duration. A possible limitation of sound-based cough frequency measurement is the contamination of recordings by environmental coughs (coughs from persons other than the patient). This study aimed to investigate the accuracy of a sound-based cough monitor for detecting and discriminating patient cough from environmental cough...
November 2016: Journal of Thoracic Disease
https://www.readbyqxmd.com/read/28058690/functions-of-chromophore-containing-domain-in-the-large-linker-lcm-polypeptide-of-phycobilisome
#3
I V Elanskaya, I A Kononova, E P Lukashev, Y V Bolychevtseva, M F Yanushin, I N Stadnichuk
In the large linker ArcE polypeptide of the phycobilisome (PBS) from the cyanobacterium Synechocystis sp. PCC 6803, the chromophore-containing 26-kDa domain was deleted with consequent disturbance of the main PBS functions. Phycobilisomes in mutant cells staying in contact with photosystem I cannot transfer energy to the photosystem II. Under the bright light conditions, the interaction of PBSs with the photoprotective orange carotenoid protein in the mutant was lost and the implementation of transition states 1 and 2 of the pigment apparatus was significantly reduced...
November 2016: Doklady. Biochemistry and Biophysics
https://www.readbyqxmd.com/read/28057717/pathology-driven-comprehensive-proteomic-profiling-of-the-prostate-cancer-tumour-microenvironment
#4
Lisa Staunton, Claire Tonry, Rosina Lis, Virginia Espina, Lance Liotta, Rosanna Inziatari, Michaela Bowden, Aurelie Fabre, John O'Leary, Stephen P Finn, Massimo Loda, Stephen R Pennington
: Prostate cancer (PCa) is the second most common cancer in men worldwide. Gleason grading is an important predictor of PCa outcomes and is influential in determining patient treatment options. Clinical decisions based on a Gleason score of 7 are difficult as the prognosis for individuals diagnosed with Gleason 4+3 cancer is much worse than for those diagnosed with Gleason 3+4 cancer. Laser capture microdissection (LCM) is a highly precise method to isolate specific cell populations or discrete microregions from tissues...
January 5, 2017: Molecular Cancer Research: MCR
https://www.readbyqxmd.com/read/28029219/defining-the-tipping-point-in-surgical-performance-for-laparoscopic-donor-nephrectomy-among-transplant-surgery-fellows-a-risk-adjusted-cumulative-summation-learning-curve-analysis
#5
O K Serrano, A S Bangdiwala, D M Vock, D Berglund, T B Dunn, E B Finger, T L Pruett, A J Matas, R Kandaswamy
UNOS recommends that fellowship-trained surgeons participate in 15 LDN to be considered proficient. ASTS mandates 12 LDN during abdominal transplant surgery fellowship (ATSF). We performed a retrospective intraoperative case analysis to create a risk-adjusted cumulative summation (RACUSUM) model to assess the learning curve (LC) of novice TSF (nTSF). Between January 2000 and December 2014, 30 nTSF rotated on the Organ Procurement Rotation (OPR) of our ASTS-approved ATSF. Measures of surgical performance included: 1) intraoperative time (ORT), 2) estimated blood loss (EBL), and 3) incidence of intraoperative complications (IOC)...
December 28, 2016: American Journal of Transplantation
https://www.readbyqxmd.com/read/28029204/translating-thermal-response-of-triblock-copolymer-assemblies-in-dilute-solution-to-macroscopic-gelation-and-phase-separation
#6
Zhe Sun, Ye Tian, Wendy L Hom, Oleg Gang, Surita R Bhatia, Robert B Grubbs
The thermal response of semi-dilute solutions (5 w/w%) of two amphiphilic thermoresponsive poly(ethylene oxide)-b-poly(N,N-diethylacrylamide)-b-poly(N,N-dibutylacrylamide) (PEO45 -PDEAmx -PDBAm12 ) triblock copolymers, which differ only in the size of the central responsive block, in water was examined. Aqueous PEO45 -PDEAm41 -PDBAm12 solutions, which undergo a thermally induced sphere-to-worm transition in dilute solution, were found to reversibly form soft (G'≈10 Pa) free-standing physical gels after 10 min at 55 °C...
December 28, 2016: Angewandte Chemie
https://www.readbyqxmd.com/read/28027905/notochord-isolation-using-laser-capture-microdissection
#7
R G C Santegoeds, Y Yakkioui, A Jahanshahi, G Raven, J J Van Overbeeke, A Herrler, Y Temel
BACKGROUND: Chordoma are malignant tumors of the axial skeleton, which arise from remnants of the notochord. The Notochord (chorda dorsalis) is an essential embryonic structure involved in the development of the nervous system and axial skeleton. Therefore, the notochord seems to be the most appropriate control tissue in chordoma molecular biology research. Nevertheless, up to now mainly different tissues but not the notochord have been used as control for chordoma, due to difficulty of isolating notochordal tissue...
December 24, 2016: Journal of Chemical Neuroanatomy
https://www.readbyqxmd.com/read/28018990/the-tcf1-bcl6-axis-counteracts-type-i-interferon-to-repress-exhaustion-and-maintain-t-cell-stemness
#8
Tuoqi Wu, Yun Ji, E Ashley Moseman, Haifeng C Xu, Monica Manglani, Martha Kirby, Stacie M Anderson, Robin Handon, Elizabeth Kenyon, Abdel Elkahloun, Weiwei Wu, Philipp A Lang, Luca Gattinoni, Dorian B McGavern, Pamela L Schwartzberg
During chronic viral infections and in cancer, T cells become dysfunctional, a state known as T cell exhaustion. Although it is well recognized that memory CD8 T cells account for the persistence of CD8 T cell immunity after acute infection, how exhausted T cells persist remains less clear. Using chronic infection with lymphocytic choriomeningitis virus clone 13 and tumor samples, we demonstrate that CD8 T cells differentiate into a less exhausted TCF1(high) and a more exhausted TCF1(low) population. Virus-specific TCF1(high) CD8 T cells, which resemble T follicular helper (TFH) cells, persist and recall better than do TCF1(low) cells and act as progenitor cells to replenish TCF1(low) cells...
December 23, 2016: Science Immunology
https://www.readbyqxmd.com/read/28005051/-russian-experience-of-lacosamide-vimpat-usage-in-treatment-of-uncontrolled-focal-epilepsy
#9
A V Lebedeva, S G Burd, O V Belyaev, P N Vlasov, M Yu Dorofeeva, A M Ismailov, M Ya Kissin, I Yu Kovaleva, N Yu Koroleva, L V Lipatova, K Yu Mukhin, Yu V Solomatin, A Chervyakov, R G Shikkherimov, O A Pylaeva, N V Freidkova
: Many patients with epilepsy receive treatment in polytherapy. Selection of antiepileptic drugs (AEDs) for the combination should be carried out in accordance with the principles of rational polytherapy, taking into account the mechanism of action, pharmacokinetic (PK) and pharmacodynamic (PD) properties of drugs. Along with levetiracetam, gabapentin, vigabatrin and pregabalin, lacosamide (LCM) shows superior PK profile in rating of all AED and can be combined with any of them. The goal of this study was to evaluate efficacy and tolerability of LCM in patients with uncontrolled partial onset seizures (POS) in routine clinical practice...
2016: Zhurnal Nevrologii i Psikhiatrii Imeni S.S. Korsakova
https://www.readbyqxmd.com/read/28004771/spatially-resolved-proteomics-rapid-quantitative-analysis-of-laser-capture-microdissected-alveolar-tissue-samples
#10
Geremy Clair, Paul D Piehowski, Teodora Nicola, Joseph A Kitzmiller, Eric L Huang, Erika M Zink, Ryan L Sontag, Daniel J Orton, Ronald J Moore, James P Carson, Richard D Smith, Jeffrey A Whitsett, Richard A Corley, Namasivayam Ambalavanan, Charles Ansong
Laser capture microdissection (LCM)-enabled region-specific tissue analyses are critical to better understand complex multicellular processes. However, current proteomics workflows entail several manual sample preparation steps and are challenged by the microscopic mass-limited samples generated by LCM, impacting measurement robustness, quantification and throughput. Here, we coupled LCM with a proteomics workflow that provides fully automated analysis of proteomes from microdissected tissues. Benchmarking against the current state-of-the-art in ultrasensitive global proteomics (FASP workflow), our approach demonstrated significant improvements in quantification (~2-fold lower variance) and throughput (>5 times faster)...
December 22, 2016: Scientific Reports
https://www.readbyqxmd.com/read/28003483/adenovirus-serotype-5-vaccination-results-in-suboptimal-cd4-t-helper-1-responses-in-mice
#11
Junghwa Lee, Masao Hashimoto, Se Jin Im, Koichi Araki, Hyun-Tak Jin, Carl W Davis, Bogumila T Konieczny, Gregory A Spies, M Juliana McElrath, Rafi Ahmed
: Adenovirus serotype 5 (Ad5) is one of the most widely used viral vectors and is known to generate potent T cell responses. While many previous studies have characterized Ad5-induced CD8 T cell responses, there is a relative lack of detailed studies analysing CD4 T cells elicited by Ad5 vaccination. Herein, we immunized mice with Ad5 vectors encoding lymphocytic choriomeningitis virus (LCMV)-glycoprotein (GP) and examined GP-specific CD4 T cell responses elicited by Ad5 vectors compared to those induced by an acute LCMV infection...
December 21, 2016: Journal of Virology
https://www.readbyqxmd.com/read/28003378/chronic-type-i-ifn-is-sufficient-to-promote-immunosuppression-through-accumulation-of-myeloid-derived-suppressor-cells
#12
Kahina Taleb, Cédric Auffray, Pascale Villefroy, Adrien Pereira, Anne Hosmalin, Muriel Gaudry, Agnès Le Bon
Failure of the immune system to eradicate viruses results in chronic viral infections, which are associated with increased susceptibility to secondary infections. Pathogenic HIV or lymphocytic choriomeningitis virus chronic infections display a persistent type I IFN signature. In chronic lymphocytic choriomeningitis virus infection, blockade of type I IFN signaling partially restores antiviral responses. In a mouse model, we tested whether chronic administration of type I IFN, at doses mimicking chronic viral infection, induced immunosuppression...
December 21, 2016: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/27994594/type-i-interferon-impairs-specific-antibody-responses-early-during-establishment-of-lcmv-infection
#13
Matthieu Daugan, Armstrong Murira, Barbara C Mindt, Amélie Germain, Esther Tarrab, Pascal Lapierre, Jörg H Fritz, Alain Lamarre
Elicitation of type I interferon (IFN-I) has been shown to both enhance and impair cell-mediated immune responses in acute and persistent viral infections, respectively. Here, we show that, in addition to its effect on T cells, IFN-I drives impairment of specific antibody responses through interaction with B cells in the acute phase of lymphocytic choriomeningitis virus (LCMV) infection. This impairment was limited to the T cell-dependent B cell response and was associated with disruption of B cell follicles, development of hypergammaglobulinemia (HGG), and expansion of the T follicular helper cell population...
2016: Frontiers in Immunology
https://www.readbyqxmd.com/read/27984679/making-the-cut-innovative-methods-for-optimizing-perfusion-based-migration-assays
#14
Andrew W Holt, William E Howard, Elizabeth T Ables, Stephanie M George, Cindy A Kukoly, Jake E Rabidou, Jake T Francisco, Angel N Chukwu, David A Tulis
Application of fluid shear stress to adherent cells dramatically influences their cytoskeletal makeup and differentially regulates their migratory phenotype. Because cytoskeletal rearrangements are necessary for cell motility and migration, preserving these adaptations under in vitro conditions and in the presence of fluid flow are physiologically essential. With this in mind, parallel plate flow chambers and microchannels are often used to conduct in vitro perfusion experiments. However, both of these systems currently lack capacity to accurately study cell migration in the same location where cells were perfused...
December 16, 2016: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/27974559/mtor-promotes-anti-viral-humoral-immunity-by-differentially-regulating-cd4-helper-t-cell-and-b-cell-responses
#15
Lilin Ye, Junghwa Lee, Lifan Xu, Ata-Ur-Rasheed Mohammed, Weiyan Li, J Scott Hale, Wendy G Tan, Tuoqi Wu, Carl W Davis, Rafi Ahmed, Koichi Araki
: mTOR has important roles in regulation of both innate and adaptive immunity but whether and how mTOR modulates humoral immune responses is yet to be fully understood. To address this issue, we examined the effects of rapamycin, a specific inhibitor of mTOR, on B cell and CD4 T cell responses during acute infection with lymphocytic choriomeningitis virus. Rapamycin treatment resulted in suppression of virus-specific B cell responses by inhibiting proliferation of germinal center (GC) B cells...
December 14, 2016: Journal of Virology
https://www.readbyqxmd.com/read/27959449/mir-204-regulates-epithelial-mesenchymal-transition-by-targeting-sp1-in-the-tubular-epithelial-cells-after-acute-kidney-injury-induced-by-ischemia-reperfusion
#16
Shun-Jie Chen, Ping Wu, Li-Jing Sun, Bo Zhou, Wei Niu, Shuang Liu, Fu-Jun Lin, Geng-Ru Jiang
Acute kidney injury (AKI) is a disease where kidney function is lost almost instantaneously; it can develop very rapidly over few hours to maximum of few days. Despite the advent of technology, the clinical management against this disease is very poor, and most of the time it is life-threatening. AKI has been actively regulated by extracellular matrix proteins (ECM), however, its underlying mechanism of regulation during AKI progression is very poorly understood. In this study, we explored the integrated network of mRNA and microRNAs (miRNAs) that maintains the progression of ECM after induction of AKI by lethal ischemia...
February 2017: Oncology Reports
https://www.readbyqxmd.com/read/27940191/the-power-of-flash-mob-research-conducting-a-nationwide-observational-clinical-study-on-capillary-refill-time-in-a-single-day
#17
Jelmer Alsma, Jan Lcm van Saase, Prabath Wb Nanayakkara, Wem Ineke Schouten, Anique Baten, Martijn P Bauer, Frits Holleman, Jack Jm Ligtenberg, Patricia M Stassen, Karin A Kaasjager, Harm Haak, Frank H Bosch, Stephanie Ce Schuit
BACKGROUND: Capillary refill time (CRT) is a clinical test used to evaluate the circulatory status of patients, and there are various methods to assess CRT. Conventional clinical research often demands large numbers of patients, making it costly, labor-intensive and time consuming. We studied the interobserver agreement on CRT in a nationwide study using a novel methodology of research called 'Flash Mob Research' (FMR). METHODS: Physicians in the Netherlands were recruited by word-of-mouth, conventional media and social media to participate in a nationwide, single-day, "nine-to-five", multi-center, cross-sectional, observational study to evaluate CRT...
December 6, 2016: Chest
https://www.readbyqxmd.com/read/27934333/thermal-rate-coefficients-for-the-astrochemical-process-c-ch-%C3%A2-c2-h-by-ring-polymer-molecular-dynamics
#18
Sergio Rampino, Yury V Suleimanov
Thermal rate coefficients for the astrochemical reaction C + CH(+) → C2(+) + H were computed in the temperature range 20-300 K by using novel rate theory based on ring polymer molecular dynamics (RPMD) on a recently published bond-order based potential energy surface and compared with previous Langevin capture model (LCM) and quasi-classical trajectory (QCT) calculations. Results show that there is a significant discrepancy between the RPMD rate coefficients and the previous theoretical results that can lead to overestimation of the rate coefficients for the title reaction by several orders of magnitude at very low temperatures...
December 22, 2016: Journal of Physical Chemistry. A
https://www.readbyqxmd.com/read/27924510/global-protein-expression-analysis-of-molecular-markers-of-ds-1-47-a-component-of-implantation-promoting-traditional-chinese-medicine
#19
Yan-Ling Li, Xiao-Yan Zhang, Yu Leng, Yan-Li Wu, Jing Li, Yun-Xia Wu
This study investigated the molecular markers of DS-1-47, a component of an implantation- promoting traditional Chinese medicine consisting of Astragalus mongholicus, Atractylodes macrocephala, Scutellaria baicalensis and Dipsacales, in an attempt to clarify the molecular mechanism and action targets of DS-1-47. Controlled ovarian stimulation (COS) method was used to establish the implantation dysfunction models of mice. Animals were divided into normal pregnant group, COS model group and DS-1-47 group. Laser capture microdissection-double dimensional electrophoresis-mass spectrum (LCM-DE-MS) was used to analyze the uterine protein molecules that were possibly involved in the promotion of implantation...
December 2016: Journal of Huazhong University of Science and Technology. Medical Sciences
https://www.readbyqxmd.com/read/27917413/efficacy-of-s-lacosamide-in-preclinical-models-of-cephalic-pain
#20
Aubin Moutal, Nathan Eyde, Edwin Telemi, Ki Duk Park, Jennifer Y Xie, David W Dodick, Frank Porreca, Rajesh Khanna
Migraine is one of the world's most common neurological disorders. Current acute migraine treatments have sub-optimal efficacy and new therapeutic options are needed. Approaches targeting calcitonin gene related peptide (CGRP) signaling are clinically effective but small molecule antagonists have not been advanced due to toxicity. In this study, we explored the axonal growth/specification collapsin response mediator protein 2 (CRMP2) as a novel "druggable" target for inhibiting CGRP release and for potential relevance for treatment of migraine pain...
June 2016: Pain Reports (Baltimore, Md.)
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