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Riti Sharan, Hee-Jeong Yang, Preeti Sule, Jeffrey D Cirillo
Reporter enzyme fluorescence (REF) utilizes substrates that are specific for enzymes present in target organisms of interest for imaging or detection by fluorescence or bioluminescence. We utilize BlaC, an enzyme expressed constitutively by all M. tuberculosis strains. REF allows rapid quantification of bacteria in lungs of infected mice. The same group of mice can be imaged at many time points, greatly reducing costs, enumerating bacteria more quickly, allowing novel observations in host-pathogen interactions, and increasing statistical power, since more animals per group are readily maintained...
February 26, 2018: Journal of Visualized Experiments: JoVE
Holger Fehlauer, Adam L Nekimken, Anna A Kim, Beth L Pruitt, Miriam B Goodman, Michael Krieg
One central goal of mechanobiology is to understand the reciprocal effect of mechanical stress on proteins and cells. Despite its importance, the influence of mechanical stress on cellular function is still poorly understood. In part, this knowledge gap exists because few tools enable simultaneous deformation of tissue and cells, imaging of cellular activity in live animals, and efficient restriction of motility in otherwise highly mobile model organisms, such as the nematode Caenorhabditis elegans. The small size of C...
February 19, 2018: Journal of Visualized Experiments: JoVE
Madeleine S Durkee, Landon D Nash, Fatemeh Nooshabadi, Jeffrey D Cirillo, Duncan J Maitland, Kristen C Maitland
The rapid development of new optical imaging techniques is dependent on the availability of low-cost, customizable, and easily reproducible standards. By replicating the imaging environment, costly animal experiments to validate a technique may be circumvented. Predicting and optimizing the performance of in vivo and ex vivo imaging techniques requires testing on samples that are optically similar to tissues of interest. Tissue-mimicking optical phantoms provide a standard for evaluation, characterization, or calibration of an optical system...
February 12, 2018: Journal of Visualized Experiments: JoVE
Chao Xiong, Zhiquan Zhang, Gurpreet S Baht, Niccolo Terrando
Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and significantly diminish the outcome. Indeed, after routine orthopedic surgery to repair a fracture, as many as 50% of elderly patients suffer from neurologic complications like delirium. Also, the capacity to heal and regenerate tissue after surgery decreases with age, and can impact the quality of fracture repair and even osseous integration of implants. Thus, a better understanding of mechanisms that drive these age-dependent changes could provide strategic targets to minimize risk for such complications and optimize outcomes...
February 27, 2018: Journal of Visualized Experiments: JoVE
Xinyi Ding, Yuxiang Sun, Yanming Chen, Wanchuan Ding, Steven Emory, Tianhao Li, Zixing Xu, Ning Han, Jun Wang, Gang Ruan
Micellar nanocrystals (micelles with encapsulated nanocrystals) have become an emerging major class of nanobiomaterials. We describe a method of fabricating micellar nanocrystals based on combining top-down electrospray, bottom-up self-assembly, and solvent-based structure control. This method involves first using electrospray to generate uniform ultrafine liquid droplets, each of which functions as a micro-reactor in which self-assembly reaction occurs forming micellar nanocrystals, with the structures (micelle shape and nanocrystal encapsulation) controlled by the organic solvent used...
February 11, 2018: Journal of Visualized Experiments: JoVE
Meredith B Rosenthal, Carrie H Colla, Nancy E Morden, Thomas D Sequist, Alexander J Mainor, Zhonghe Li, Kevin H Nguyen
OBJECTIVES: A substantial portion of healthcare spending is wasted on services that do not directly improve patient health and that cause harm in some cases. Features of health insurance coverage, including enrollment in high-deductible health plans (HDHPs) or health maintenance organizations (HMOs), may provide financial and nonfinancial mechanisms to potentially reduce overuse of low-value healthcare services. STUDY DESIGN: Using 2009 to 2013 administrative data from 3 large commercial insurers, we examined patient characteristics and health insurance plan types associated with overuse of 6 healthcare services identified by the Choosing Wisely campaign...
March 2018: American Journal of Managed Care
Alajos Pár, Gabriella Pár
More than 25 years after the discovery of hepatitis C virus, the development of the direct acting antivirals can lead to the regional or long-term global elimination of the virus with over 90% efficacy. This is the success of basic and clinical translational research. Yet, some unsolved challanges remain, such as the great number of unidentified patients who are not aware of their condition, the limited access to the therapy due to the high prices of the drugs, and the treatment of resistance-associated variants...
March 2018: Orvosi Hetilap
Sumit Handa, Daniel R Dempsey, Divya Ramamoorthy, Nanci Cook, Wayne C Guida, Tyler J Spradling, Justin K White, H Lee Woodcock, David J Merkler
The non-mevalonate dependent (NMVA) pathway for the biosynthesis of isopentenyl pyrophosphate and dimethylallyl pyrophosphate is the sole source of these terpenoids for the production of isoprenoids in the apicomplexan parasites, in many eubacteria, and in plants. The absence of this pathway in higher organisms has opened a new platform for the development of novel antibiotics and anti-malarials. The enzyme catalyzing the first step of the NMVA pathway is 1-deoxy-D-xylulose-5-phosphate synthase (DXPS). DXPS catalyzes the thiamine pyrophosphate- and Mg (II)-dependent conjugation of pyruvate and D-glyceraldehyde-3-phosphate to form 1-deoxy-D-xylulose-5-phosphate and CO2 ...
2018: Biochemistry & Molecular Biology Journal
John A Calarco, Adam D Norris
Genetic interaction screens are a powerful methodology to establish novel roles for genes and elucidate functional connections between genes. Such studies have been performed to great effect in single-cell organisms such as yeast and E. coli (Schuldiner et al ., 2005; Butland et al ., 2008; Costanzo et al ., 2010), but similar large-scale interaction studies using targeted reverse-genetic deletions in multi-cellular organisms have not been feasible. We developed a CRISPR/Cas9-based method for deleting genes in C...
March 5, 2018: Bio-protocol
Syed M Hasan, Nida N Ahmed, Zunirah Ahmed, Allan Seibert
Amyloidosis is a rare disorder with a wide spectrum of presentations and anomalies. It is subdivided into 2 broad categories based on protein deposition; primary and secondary amyloidosis. It can present as a single-organ involvement or as a diffuse infiltrative multi-organ process. Isolated hepatic amyloidosis presentation is a rare phenomenon that develops due to insoluble amyloid deposition in liver. Its clinical presentation is usually vague and ranges from mild hepatomegaly with elevated liver enzymes to acute liver failure and hepatic rupture...
January 2018: Journal of Investigative Medicine High Impact Case Reports
Antonia Reimer, Yinghong He, Cristina Has
Genetic conditions affecting the skin and kidney are clinically and genetically heterogeneous, and target molecular components present in both organs. The molecular pathology involves defects of cell-matrix adhesion, metabolic or signaling pathways, as well as tumor suppressor genes. This article gives a clinically oriented overview of this group of disorders, highlighting entities which have been recently described, as well as the progress made in understanding well-known entities. The genetic bases as well as molecular cell biological mechanisms are described, with therapeutic applications...
2018: Frontiers in Pediatrics
Jacob D Estes, Roger LeGrand, Constantinos Petrovas
Immunological inductive tissues, such as secondary lymphoid organs, are composed of distinct anatomical microenvironments for the generation of immune responses to pathogens and immunogens. These microenvironments are characterized by the compartmentalization of highly specialized immune and stromal cell populations, as well as the presence of a complex network of soluble factors and chemokines that direct the intra-tissue trafficking of naïve and effector cell populations. Imaging platforms have provided critical contextual information regarding the molecular and cellular interactions that orchestrate the spatial microanatomy of relevant cells and the development of immune responses against pathogens...
2018: Frontiers in Immunology
Jiadela Teliewubai, Bin Bai, Yiwu Zhou, Yuyan Lu, Shikai Yu, Chen Chi, Jue Li, Jacques Blacher, Yawei Xu, Yi Zhang
Objective: Secreted frizzled related protein 5 (SFRP5) is a novel anti-inflammatory adipokine that is implicated in metabolic and cardiovascular disease (CVD). However, little is known about the relevance of SFRP5 with asymptomatic hypertensive target organ damages (TODs). We aimed to investigate the association between SFRP5 and TOD in a large population. Clinical trial registration: NCT02368938. Methods: A total of 1,745 community-dwelling elderly subjects aged over 65 years from northern Shanghai were recruited in the study...
2018: Clinical Interventions in Aging
Tsutomu Nakamura, Daisuke Koma, Maki Oshima, Hideto Hoshino, Takashi Ohmoto, Koichi Uegaki
Escherichia coli is the most popular organism used for producing recombinant proteins. However, the expression of recombinant proteins in E. coli sometimes results in the aggregation of proteins as an inclusion body in host cells. In such cases, it is necessary to optimize the refolding conditions to obtain the recombinant protein in its native form. Several techniques, such as reducing the concentration of the induction reagent during E. coli cultivation, have been developed to prevent the formation of inclusion bodies by controlling protein expression levels...
March 15, 2018: Journal of Bioscience and Bioengineering
Mauro E M Nunes, Talise E Müller, Camila Murussi, Aline M B Amaral, Jeane L C Gomes, Aline T Marins, Jossiele Leitemperger, Cintia C R Rodrigues, Tiago L Fiuza, Maiara D Costa, Eduardo S Severo, Denis B Rosemberg, Vania L Loro
The use of commercial pesticides combinations increases the risk of intoxication in non-target aquatic organisms. Here, we investigate the potential of a commercial pesticide formulation containing (CYP) plus chlopyrifos (CPF) to induce oxidative damage on two fish species (common carp and zebrafish). Carp and zebrafish were exposed for 96 h under laboratory conditions. Fish were divided in three different groups: CTL, 0.3 μg L-1 or 0.6 μg L-1 of CYP and 0.5 or 1 μg L-1 of CPF in commercial formulation...
March 15, 2018: Comparative Biochemistry and Physiology. Toxicology & Pharmacology: CBP
Paola Fincheira, Andrés Quiroz
Agricultural practices require novel products that allow sustainable development and commercial production according to the needs of farmers and consumers. Therefore, in the last decade, eco-friendly alternatives have been studied, so volatile organic compounds (VOCs) emitted by microorganisms have emerged as a cheaper, effective, efficient, and an eco-friendly alternative. VOCs are lipophilic compounds derived from microbial metabolic pathways with low molecular weight (<300 g mol-1 ), low boiling point, and high vapor pressure that allow them to act as signal molecules over short and long distances...
March 2018: Microbiological Research
Dandan Han, Lanying Wang, Yanping Luo
Actinomycetes are an important group of gram-positive bacteria that play an essential role in the rhizosphere ecosystem. The confrontation culture and Oxford cup method were used to evaluate the antagonistic activities of strains, which were isolated from the rhizosphere soil of Mikania micrantha. The two isolates were identified using morphological and physiological tests combined with 16S rRNA-based molecular analysis, respectively. The type I polyketone synthase (PKS-I) was amplified. The constituents of fermentation metabolites were analyzed by gas chromatography mass spectrometry...
March 2018: Microbiological Research
Davar Abedini, Sajad Rashidi Monfared
A primary mechanism for controlling the development of multicellular organisms is transcriptional regulation, which carried out by transcription factors (TFs) that recognize and bind to their binding sites on promoter region. The distance from translation start site, order, orientation, and spacing between cis elements are key factors in the concentration of active nuclear TFs and transcriptional regulation of target genes. In this study, overrepresented motifs in cold and pathogenesis responsive genes were scanned via Gibbs sampling method, this method is based on detection of overrepresented motifs by means of a stochastic optimization strategy that searches for all possible sets of short DNA segments...
March 17, 2018: Molecular Biology Reports
Vadim V Chumak, Sergiy V Klymenko, Horst Zitzelsberger, Christina Wilke, Lyudmila A Rybchenko, Elena V Bakhanova
The Chernobyl reactor accident in 1986 has caused significant exposure to ionizing radiation of the Ukrainian population, in particular clean-up workers and evacuees from the exclusion zones. A study aiming at the discovery of radiation markers of the breast cancer was conducted from 2008 to 2015 within a collaborative project by HZM, LMU, and NRCRM. In this study, post-Chernobyl breast cancer cases both in radiation-exposed female patients diagnosed at age less than 60 from 1992 to 2014 and in non-exposed controls matched for residency, tumor type, age at diagnosis, TNM classification as well as tumor grading were investigated for molecular changes with special emphasis to copy number alterations and miRNA profiles...
March 17, 2018: Radiation and Environmental Biophysics
Filipa Caeiro Alves, Rute Aguiar, Pedro Pessegueiro, Carlos Pires
Thrombotic microangiopathy (TMA) comprises a process of sequential endothelial damage, microvascular thrombosis, consumptive thrombocytopenia and microangiopathic haemolytic anaemia that can affect several organs, including the kidney. A 36-year-old woman was presented with a petechial rash 3 weeks after an upper respiratory tract infection. Laboratory results showed normocytic normochromic anaemia, thrombocytopenia and evidence of TMA with decreased haptoglobin, elevated serum lactate dehydrogenase and a peripheral blood smear with numerous schistocytes...
March 17, 2018: BMJ Case Reports
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